Abstract
Twenty bacterial strains were isolated on selective milk agar plates (pH 9.0) from tannery solid waste on the basis of different
colony morphology. These strains exhibited variable haloalkaline protease activity and were tolerant to different concentration of
both chromate (350-1450 μg/ml) and NaCl (2-9%). Those having clearance zone greater than 20.0 mm were considered as
significant isolate. Out of twenty, nine strains were interestingly tolerant to high concentration of Cr(VI) (850-1450 mg/l) and
NaCl (6.5-9%) and exhibited vibrant clear zone diameter between 21-35 mm. All these isolates in this study were capable of
reducing Cr(VI) aerobically and the reduction values ranged between 50.0-76.0% after 16-20 h of growth. Further, significant
haloalkaline protease production was observed in these bacteria within 24 h under wide temperature (25-45°C) and pH (8.0-10.5)
range. The potential of these strains to produce protease at higher rate in the presence of wheat bran as a cheap carbon source
and yeast extract or beef extract as a nitrogen source makes them a potential candidate for industrial applications and removal of
Cr(VI) and proteinaceous waste simultaneously from industrial waste of alkaline nature.
Key words: Tannery waste, chromate resistant, bacteria, haloalkaline, protease
Microbial Production Of Alkaline Proteases And Evaluation Of Its Performances...Shafkat Shamim Rahman
A high alkaline protease producing bacterial strain was isolated and identified a local soil sample. The organism was gram positive and forms spore during adverse condition in the growth medium. After various tests it was suggested and the features agreed with the description of Bacillus subtilis. It was also identified as B. subtilis with 99.9% identity by API 50 CHB. The enzyme hydrolyses a number of proteins including azocasein which suggests that it is an extracellular alkaline protease. The experimentally determined isoelectric point was 5.1 and the optimal enzyme activity was at 60°C and at pH 8.5. The esterase preferentially hydrolyzed short-chain fatty acids. Native enzyme preparations typically showed a Michaelis constant (Km) and Vmax of 0.40mM and 12,200 U mg)-1, respectively. This microbial enzyme was partially purified by ammonium sulfate fractionation, dialysis, DEAE cellulose chromatography and electrophoretic analysis. Enzyme purity was tested by SDS-PAGE. Quantitative estimation has shown that 40mL of culture supernatant could dehair 2×1 cm of leather completely in 9 hours. In future the tanneries will use a combination of chemical and enzymatic processes. In practical applications, protease is a useful enzyme for promoting the hydrolysis of proteins and showing significant industrial applications.
Proteases are protein-degrading enzymes that catalyses hydrolytic reaction in which protein molecules are degraded into peptides and amino acids. Thermostable alkaline proteases are of particular great interest for industrial application because they are stable and active at temperature above 60-70˚C. Thermophiles are found in wide array of environment such as mushroom compost material, nest, hay, wood chips, grains, soil, manure, coal mines etc. Alkaline proteases are most important industrial enzymes and they occupy about 60% of total enzyme market. From the soil samples, eight different fungal species were isolated through soil dilution plate method. In the present study, two fungi Aspergillus nidulans and Aspergillus glaucus from mushroom compost and two fungi Aspergillus terrus, and Aspergillus fumigates from cow manure, showing alkaline protease activity, were isolated. The zones of clearance were observed in Aspergillus nidulans, Aspergillus glaucus, Aspergillus terrus, and Aspergillus fumigatus species of fungi isolated from cow manure and mushroom compost. The best enzyme production was observed in Aspergillus terrus (1.005 ± 0.057 IU/mg protein) obtained from cow manure and the minimum enzyme activity was observed with Aspergillus glaucus (0.278 ± 0.026 IU/mg protein). However, more studies are required to assess the potential of Aspergillus nidulans, Aspergillus glaucus, Aspergillus terrus, and Aspergillus fumigatus species. Key-words- Alkaline protease, Thermophiles, Zone of clearance, Trichloroacetic acid
PRODUCTION OF PROTEASE BY ALKALOPHILIC BACILLUSSUBTILIS IN BIOREACTOR AND ITS...AM Publications
The current studies were aimed at to investigate role of pH, dissolved oxygen for production protease in bioreactor by alkalophilic bacterium and application of saw dust for its purification. The production of proteolytic enzyme by Bacillus subtilis IC-5 started as pH of medium falls to 9 and reached to maximum at pH 7 i.e., 4400 Uml-1 . Likewise dissolved oxygen decreased in the medium as the protease production progresses. Saw dust was successively utilized for partial purification of protease. The partial purification of protease increased the specific activity to5.3 fold. The optimum pH and temperature for purified activity was 11 and 700C, respectively. The purified enzyme was stable up to pH 12 and 80oC.
Alkaline Protease - One of the class of protease enzyme.
An extracellular enzyme.
Performs proteolysis, that is, protein catabolism by hydrolysis of the peptide bonds.
Active at alkaline pH 8 to 12 and at temperature 30⁰-80⁰C.
Molecular weight is about 20,000 to 45,000 Dalton.
The structure is determined by X-ray crystallography.
EC (Enzyme Commission) Number: 3.4.21–24.99
In 1971, Japanese scientist Koki Horikoshi first reported the production of alkaline protease from bacteria.
Production and optimization of lipase from candida rugosa using groundnut oil...eSAT Publishing House
IJRET : International Journal of Research in Engineering and Technology is an international peer reviewed, online journal published by eSAT Publishing House for the enhancement of research in various disciplines of Engineering and Technology. The aim and scope of the journal is to provide an academic medium and an important reference for the advancement and dissemination of research results that support high-level learning, teaching and research in the fields of Engineering and Technology. We bring together Scientists, Academician, Field Engineers, Scholars and Students of related fields of Engineering and Technology.
Microbial Production Of Alkaline Proteases And Evaluation Of Its Performances...Shafkat Shamim Rahman
A high alkaline protease producing bacterial strain was isolated and identified a local soil sample. The organism was gram positive and forms spore during adverse condition in the growth medium. After various tests it was suggested and the features agreed with the description of Bacillus subtilis. It was also identified as B. subtilis with 99.9% identity by API 50 CHB. The enzyme hydrolyses a number of proteins including azocasein which suggests that it is an extracellular alkaline protease. The experimentally determined isoelectric point was 5.1 and the optimal enzyme activity was at 60°C and at pH 8.5. The esterase preferentially hydrolyzed short-chain fatty acids. Native enzyme preparations typically showed a Michaelis constant (Km) and Vmax of 0.40mM and 12,200 U mg)-1, respectively. This microbial enzyme was partially purified by ammonium sulfate fractionation, dialysis, DEAE cellulose chromatography and electrophoretic analysis. Enzyme purity was tested by SDS-PAGE. Quantitative estimation has shown that 40mL of culture supernatant could dehair 2×1 cm of leather completely in 9 hours. In future the tanneries will use a combination of chemical and enzymatic processes. In practical applications, protease is a useful enzyme for promoting the hydrolysis of proteins and showing significant industrial applications.
Proteases are protein-degrading enzymes that catalyses hydrolytic reaction in which protein molecules are degraded into peptides and amino acids. Thermostable alkaline proteases are of particular great interest for industrial application because they are stable and active at temperature above 60-70˚C. Thermophiles are found in wide array of environment such as mushroom compost material, nest, hay, wood chips, grains, soil, manure, coal mines etc. Alkaline proteases are most important industrial enzymes and they occupy about 60% of total enzyme market. From the soil samples, eight different fungal species were isolated through soil dilution plate method. In the present study, two fungi Aspergillus nidulans and Aspergillus glaucus from mushroom compost and two fungi Aspergillus terrus, and Aspergillus fumigates from cow manure, showing alkaline protease activity, were isolated. The zones of clearance were observed in Aspergillus nidulans, Aspergillus glaucus, Aspergillus terrus, and Aspergillus fumigatus species of fungi isolated from cow manure and mushroom compost. The best enzyme production was observed in Aspergillus terrus (1.005 ± 0.057 IU/mg protein) obtained from cow manure and the minimum enzyme activity was observed with Aspergillus glaucus (0.278 ± 0.026 IU/mg protein). However, more studies are required to assess the potential of Aspergillus nidulans, Aspergillus glaucus, Aspergillus terrus, and Aspergillus fumigatus species. Key-words- Alkaline protease, Thermophiles, Zone of clearance, Trichloroacetic acid
PRODUCTION OF PROTEASE BY ALKALOPHILIC BACILLUSSUBTILIS IN BIOREACTOR AND ITS...AM Publications
The current studies were aimed at to investigate role of pH, dissolved oxygen for production protease in bioreactor by alkalophilic bacterium and application of saw dust for its purification. The production of proteolytic enzyme by Bacillus subtilis IC-5 started as pH of medium falls to 9 and reached to maximum at pH 7 i.e., 4400 Uml-1 . Likewise dissolved oxygen decreased in the medium as the protease production progresses. Saw dust was successively utilized for partial purification of protease. The partial purification of protease increased the specific activity to5.3 fold. The optimum pH and temperature for purified activity was 11 and 700C, respectively. The purified enzyme was stable up to pH 12 and 80oC.
Alkaline Protease - One of the class of protease enzyme.
An extracellular enzyme.
Performs proteolysis, that is, protein catabolism by hydrolysis of the peptide bonds.
Active at alkaline pH 8 to 12 and at temperature 30⁰-80⁰C.
Molecular weight is about 20,000 to 45,000 Dalton.
The structure is determined by X-ray crystallography.
EC (Enzyme Commission) Number: 3.4.21–24.99
In 1971, Japanese scientist Koki Horikoshi first reported the production of alkaline protease from bacteria.
Production and optimization of lipase from candida rugosa using groundnut oil...eSAT Publishing House
IJRET : International Journal of Research in Engineering and Technology is an international peer reviewed, online journal published by eSAT Publishing House for the enhancement of research in various disciplines of Engineering and Technology. The aim and scope of the journal is to provide an academic medium and an important reference for the advancement and dissemination of research results that support high-level learning, teaching and research in the fields of Engineering and Technology. We bring together Scientists, Academician, Field Engineers, Scholars and Students of related fields of Engineering and Technology.
The International Journal of Engineering and Science (The IJES)theijes
The International Journal of Engineering & Science is aimed at providing a platform for researchers, engineers, scientists, or educators to publish their original research results, to exchange new ideas, to disseminate information in innovative designs, engineering experiences and technological skills. It is also the Journal's objective to promote engineering and technology education. All papers submitted to the Journal will be blind peer-reviewed. Only original articles will be published.
Production of secondary metabolites : enzymes which involves the upstream technological process
Introduction
History
Process involved
Contribution of different micro-organisms
Flowchart
Example: Methods Production of Amyalse in industrial view
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is an open access international journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Production and Purification of Amylase from Bacillus subtilis Isolated from SoilDr. Amarjeet Singh
In spite of progress in biotechnology and
enzymology, the enzymes have been industrialized in recent
years for the mounting up the product development in
various arena. The ultimate goal of this study comprises the
production and purification the amylase enzyme from the
bacterial strain. A powerful amylase producer, Bacillus
subtilis ISOLATE-4 was isolated, screened and identified
from the soil sample. In order to produce extracellular
amylase, various physico-chemical parameters were
optimized. During optimization, the maximal production of
amylase by the isolate at 48 hrs of incubation in 100 rpm was
found to be 6.93U/ml, 5.94U/ml, 6.0U/ml at 45ºC, pH 6 with
1% substrate concentration respectively. Ammonium
sulphate fractionation was done for rapid precipitation of the
amylase at a concentration of 60% and exposed to dialysis
showed the 25% purification fold of an enzyme. The dialyzed
product was further subjected to DEAE-Cellulose column
chromatography resulted in an increase up to 75%
purification fold than crude enzyme. The amylase enzyme
might be suitable for the liquefaction of starch, detergent,
textile and several additional industrial applications.
Amylase is an enzyme that catalyzes the hydrolysis of starch into sugars.
Amylase is present in the saliva of humans and some other mammals, where it begins the chemical process of digestion. Foods that contain large amount of starch but less amount of sugar, such as rice and potatoes, may acquire a slightly sweet taste as they are chewed because amylase degrades some of their starch into sugar.
α- amylase is a protein enzyme that hydrolyses alpha bonds of large, alpha - linked polysaccharides, such as starch and glycogen, yielding glucose and maltose. It is a major form of amylase found in humans and other mammals.
Many of the enzymes used in the industries are extracellular derived from microorganisms. Among various extracellular enzymes, alpha amylase ranks first in terms of commercial exploitation.
Bacteria and fungi secrets amylases to the outside of the cells to carryout extracellular digestions when they have broken down the soluble starch, the soluble end products such as Glucose or Maltose are absorbed into their cells.
The industrially important Bacillus strains which are extensively used to produce alpha amylase, are, B. licheniformis, B. subtilis etc. B. amyloliquefaciens
Bacillus licheniformis is a Gram-positive endospore forming organism that can be isolated from soils and plant material all over the world.
This organism is used extensively for large-scale industrial production of exoenzymes as it can secrete large quantities of proteins of up to 20–25 g/l.
The use of the submerged culture is advantageous because of the ease of sterilization and its process control.
The objective of this work was to study the pattern and the comparison of α-amylase production by using two strains of Bacillus licheniformis, MTCC 2617 and MTCC 2618 using four different substrates starch, rice, wheat and ragi powder as carbon source.
Ragi or finger millet is round, soft yet firm and rich brown in color. It is probably the only edible solid you are advised to swallow not chew. A gram of ragi has 72% carbohydrate, 3.6% fiber, 7.3% of protein, vitamin B and a good combination of minerals.
Production of cellulase and it's applicationRezwana Nishat
Cellulase is an enzyme that are found in livestock animals and herbivores' digestive system. It is also found in microbes system which is a great deal for researchers to study this enzymatic system furthermore. In this presentation, the production and the applications of this enzyme for biostoning of denim and cellulose nanofiber production have been studied.
Isolation and purification of peroxidase from soyabeanPooja Walke
Peroxidase (EC. 1.11.1.7), an oxidoreductase, has iron porphyrin ring generally and catalyzes a redox reaction between H202 as an electron acceptor and many kinds of substrates by means of oxygen liberation from HzOz (Brill, 1996).
ABSTRACT- Microbial source of amylase is preferred to other sources because of its plasticity, vast availability, higher yield and
thermostability even at elevated temperatures.Various physical and chemical factors have been known to affect the production of α-
amylase such as temperature, pH, period of incubation, carbon sources acting as inducers, surfactants, nitrogen sources, phosphate,
different metal ions, moisture. Interactions of these parameters are reported to have a significant influence on the production of
the enzyme.Study was mainly aimed to isolate a bacterium capable of hydrolyzing a starch source and to check effect of different physiological
parameters on amylase enzyme activity. To conduct this research, study was mainly focused on three objectives i.e. 1st Screening
and morphological characterization of the isolated bacteria. 2nd Characterization of amylase production by selected isolates. 3rd
Time course of Enzyme production and Partial purification with Ammonium Sulphate saturation.Amylases of isolate-6 and isolate-9
were concentrated by ammonium sulfate precipitation which can be used as partially purified enzyme for further study. Isolate-6 and
Isolate-9 showed the activity 0.34 and 0.28 units/ml/min respectively.Enzyme derived from isolate-6 and isolate-9 was stable at different
physiological conditions. So, it is useful in fermentation industry and in pharmaceuticals.
Key words- Amylase, Starch hydrolyzing bacteria, fermentation and pharmaceutical industries
UNIT 6 Fermentation technology, Fermenters, Study of Media, types of fermenta...Shyam Bass
UNIT-6 6th Sem B.Pharma Pharmaceutical Biotechnology-
Following slides include-
Fermentation technology and biotechnological products :
Fermentation methods and general requirements
Study of media
Equipment
Sterilization methods
Aeration process
Stirring
large scale production fermenter design and its various controls
BY- SHYAM BASS
The International Journal of Engineering and Science (The IJES)theijes
The International Journal of Engineering & Science is aimed at providing a platform for researchers, engineers, scientists, or educators to publish their original research results, to exchange new ideas, to disseminate information in innovative designs, engineering experiences and technological skills. It is also the Journal's objective to promote engineering and technology education. All papers submitted to the Journal will be blind peer-reviewed. Only original articles will be published.
The papers for publication in The International Journal of Engineering& Science are selected through rigorous peer reviews to ensure originality, timeliness, relevance, and readability
The International Journal of Engineering and Science (The IJES)theijes
The International Journal of Engineering & Science is aimed at providing a platform for researchers, engineers, scientists, or educators to publish their original research results, to exchange new ideas, to disseminate information in innovative designs, engineering experiences and technological skills. It is also the Journal's objective to promote engineering and technology education. All papers submitted to the Journal will be blind peer-reviewed. Only original articles will be published.
Production of secondary metabolites : enzymes which involves the upstream technological process
Introduction
History
Process involved
Contribution of different micro-organisms
Flowchart
Example: Methods Production of Amyalse in industrial view
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is an open access international journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Production and Purification of Amylase from Bacillus subtilis Isolated from SoilDr. Amarjeet Singh
In spite of progress in biotechnology and
enzymology, the enzymes have been industrialized in recent
years for the mounting up the product development in
various arena. The ultimate goal of this study comprises the
production and purification the amylase enzyme from the
bacterial strain. A powerful amylase producer, Bacillus
subtilis ISOLATE-4 was isolated, screened and identified
from the soil sample. In order to produce extracellular
amylase, various physico-chemical parameters were
optimized. During optimization, the maximal production of
amylase by the isolate at 48 hrs of incubation in 100 rpm was
found to be 6.93U/ml, 5.94U/ml, 6.0U/ml at 45ºC, pH 6 with
1% substrate concentration respectively. Ammonium
sulphate fractionation was done for rapid precipitation of the
amylase at a concentration of 60% and exposed to dialysis
showed the 25% purification fold of an enzyme. The dialyzed
product was further subjected to DEAE-Cellulose column
chromatography resulted in an increase up to 75%
purification fold than crude enzyme. The amylase enzyme
might be suitable for the liquefaction of starch, detergent,
textile and several additional industrial applications.
Amylase is an enzyme that catalyzes the hydrolysis of starch into sugars.
Amylase is present in the saliva of humans and some other mammals, where it begins the chemical process of digestion. Foods that contain large amount of starch but less amount of sugar, such as rice and potatoes, may acquire a slightly sweet taste as they are chewed because amylase degrades some of their starch into sugar.
α- amylase is a protein enzyme that hydrolyses alpha bonds of large, alpha - linked polysaccharides, such as starch and glycogen, yielding glucose and maltose. It is a major form of amylase found in humans and other mammals.
Many of the enzymes used in the industries are extracellular derived from microorganisms. Among various extracellular enzymes, alpha amylase ranks first in terms of commercial exploitation.
Bacteria and fungi secrets amylases to the outside of the cells to carryout extracellular digestions when they have broken down the soluble starch, the soluble end products such as Glucose or Maltose are absorbed into their cells.
The industrially important Bacillus strains which are extensively used to produce alpha amylase, are, B. licheniformis, B. subtilis etc. B. amyloliquefaciens
Bacillus licheniformis is a Gram-positive endospore forming organism that can be isolated from soils and plant material all over the world.
This organism is used extensively for large-scale industrial production of exoenzymes as it can secrete large quantities of proteins of up to 20–25 g/l.
The use of the submerged culture is advantageous because of the ease of sterilization and its process control.
The objective of this work was to study the pattern and the comparison of α-amylase production by using two strains of Bacillus licheniformis, MTCC 2617 and MTCC 2618 using four different substrates starch, rice, wheat and ragi powder as carbon source.
Ragi or finger millet is round, soft yet firm and rich brown in color. It is probably the only edible solid you are advised to swallow not chew. A gram of ragi has 72% carbohydrate, 3.6% fiber, 7.3% of protein, vitamin B and a good combination of minerals.
Production of cellulase and it's applicationRezwana Nishat
Cellulase is an enzyme that are found in livestock animals and herbivores' digestive system. It is also found in microbes system which is a great deal for researchers to study this enzymatic system furthermore. In this presentation, the production and the applications of this enzyme for biostoning of denim and cellulose nanofiber production have been studied.
Isolation and purification of peroxidase from soyabeanPooja Walke
Peroxidase (EC. 1.11.1.7), an oxidoreductase, has iron porphyrin ring generally and catalyzes a redox reaction between H202 as an electron acceptor and many kinds of substrates by means of oxygen liberation from HzOz (Brill, 1996).
ABSTRACT- Microbial source of amylase is preferred to other sources because of its plasticity, vast availability, higher yield and
thermostability even at elevated temperatures.Various physical and chemical factors have been known to affect the production of α-
amylase such as temperature, pH, period of incubation, carbon sources acting as inducers, surfactants, nitrogen sources, phosphate,
different metal ions, moisture. Interactions of these parameters are reported to have a significant influence on the production of
the enzyme.Study was mainly aimed to isolate a bacterium capable of hydrolyzing a starch source and to check effect of different physiological
parameters on amylase enzyme activity. To conduct this research, study was mainly focused on three objectives i.e. 1st Screening
and morphological characterization of the isolated bacteria. 2nd Characterization of amylase production by selected isolates. 3rd
Time course of Enzyme production and Partial purification with Ammonium Sulphate saturation.Amylases of isolate-6 and isolate-9
were concentrated by ammonium sulfate precipitation which can be used as partially purified enzyme for further study. Isolate-6 and
Isolate-9 showed the activity 0.34 and 0.28 units/ml/min respectively.Enzyme derived from isolate-6 and isolate-9 was stable at different
physiological conditions. So, it is useful in fermentation industry and in pharmaceuticals.
Key words- Amylase, Starch hydrolyzing bacteria, fermentation and pharmaceutical industries
UNIT 6 Fermentation technology, Fermenters, Study of Media, types of fermenta...Shyam Bass
UNIT-6 6th Sem B.Pharma Pharmaceutical Biotechnology-
Following slides include-
Fermentation technology and biotechnological products :
Fermentation methods and general requirements
Study of media
Equipment
Sterilization methods
Aeration process
Stirring
large scale production fermenter design and its various controls
BY- SHYAM BASS
The International Journal of Engineering and Science (The IJES)theijes
The International Journal of Engineering & Science is aimed at providing a platform for researchers, engineers, scientists, or educators to publish their original research results, to exchange new ideas, to disseminate information in innovative designs, engineering experiences and technological skills. It is also the Journal's objective to promote engineering and technology education. All papers submitted to the Journal will be blind peer-reviewed. Only original articles will be published.
The papers for publication in The International Journal of Engineering& Science are selected through rigorous peer reviews to ensure originality, timeliness, relevance, and readability
Phycoremediation of malachite green and reduction of physico chemical paramet...eSAT Journals
Abstract
Water is the elixir of life, a precious gift of nature to all the living species on earth. It is rapidly becoming a scare commodity in most parts of the world. Only 0.35% of the total availability of water found in lakes and wetlands and 0.01% in rivers and streams which are likely getting depleted due to the discharge of the effluents such as dyes simultaneously increases the water quality parameters. Malachite green is a common textile dye being discharged in lake water situated near textile industries. The phycoremediation method employs the use of algae, say, Chlorella pyrenoidosa which helps in decolorizing the water and due to its growth in the lake water, the physico-chemical parameters higher in the polluted water found to be reduced and water can be used for various purposes. The trails were made using Chlorella pyrenoidosa and Malachite green at different concentrations. The optimum concentration for the degradation of dye was completely done at the concentration 15 mg confirming the decolorization capacity by Chlorella pyrenoidosa. The same experiment was performed with adjusting the pH at 6, 7 and 8. Optimum pH for the dye degradation was found as pH 7 at which all the concentrations Chlorella pyrenoidosa was able to degrade the dye with the decolourization capacity of above 95%. The phyico-chemical parameters checked after the decolorization by Chlorella were found to be within the limits and thus Chlorella which is available in nature can be used in the decolorization of effluent water by eco-friendly method.
Keywords: Chlorella pyrenoidosa, Malachite Green, Phycoremediation, Physico-chemical parameters
IJRET : International Journal of Research in Engineering and Technology is an international peer reviewed, online journal published by eSAT Publishing House for the enhancement of research in various disciplines of Engineering and Technology. The aim and scope of the journal is to provide an academic medium and an important reference for the advancement and dissemination of research results that support high-level learning, teaching and research in the fields of Engineering and Technology. We bring together Scientists, Academician, Field Engineers, Scholars and Students of related fields of Engineering and Technology
Microbial Decolorization of Leather Dye Effluentijtsrd
As we know 71% of Earths Surface is covered with water. Water is one of the most essential element for the persistence of life on this planet. Pure and contamination free water is the dire necessity of every living being present on Earth but today water pollution has posed great threat to the existence of life. The discharge of effluents from various industries into the water bodies are mainly responsible for water pollution specially effluents from leather industry are the most contaminating as along with the harmful chemicals they contain organic matter as well. The dyes disposed by the leather industry are the most harmful for the environment. The conventional physicochemical methods used for the treatment of leather dyes in the contaminated water i.e. coagulation, precipitation, oxidation etc have disadvantages and limitations. This study presents microbial decolorization of leather dye effluents from Shigella sp. isolated from effluent samples collected from leather industry. Different parameters such as temperature and pH were optimized for decolorization of Methylene Blue, Crystal Violet, EosinYellow, Safranine dyes by using bacterial isolates. Optimum temperature for decolorization was observed to be 300c, the optimum pH range for decolorization was found to be from pH6-pH8. All the samples were incubated at 30-°C 150 rpm. The decolorization was measured as decrease in absorbance maxima at 663 nm, 590 nm, 518nm, 530 nm for mehtylene blue, crystal violet, eosin yellow, safranine respectively. Dr. Jitender Kumar | Navleen Kaur Chopra"Microbial Decolorization of Leather Dye Effluent" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-1 | Issue-5 , August 2017, URL: http://www.ijtsrd.com/papers/ijtsrd2405.pdf http://www.ijtsrd.com/biological-science/biotechnology/2405/microbial-decolorization-of-leather-dye-effluent/dr-jitender-kumar
International Journal of Engineering Research and Applications (IJERA) is an open access online peer reviewed international journal that publishes research and review articles in the fields of Computer Science, Neural Networks, Electrical Engineering, Software Engineering, Information Technology, Mechanical Engineering, Chemical Engineering, Plastic Engineering, Food Technology, Textile Engineering, Nano Technology & science, Power Electronics, Electronics & Communication Engineering, Computational mathematics, Image processing, Civil Engineering, Structural Engineering, Environmental Engineering, VLSI Testing & Low Power VLSI Design etc.
International Refereed Journal of Engineering and Science (IRJES)irjes
International Refereed Journal of Engineering and Science (IRJES) is a leading international journal for publication of new ideas, the state of the art research results and fundamental advances in all aspects of Engineering and Science. IRJES is a open access, peer reviewed international journal with a primary objective to provide the academic community and industry for the submission of half of original research and applications
Multidrug resistance pattern of bacteria isolated from domestic and tannery w...iosrjce
The objective of this study was to screen for the microorganisms in the domestic wastes and tannery
effluents and characterize it’s antibiogram to know the prevalence of resistant pathogens. A total of 109 isolates
of 16 different genera were isolated from 38 samples by aerobic culture method. Fourteen and ninety-five
isolates were obtained from six domestic wastes and 32 tannery waste samples, respectively. The isolates
belonged to the genus Micrococcus (18.3%), Alcaligens (15.6%), Staphylococcus (11.0%), Enterobacter (4.6%),
Shigella (14.7%), Klebsiella (6.4%), Haemophilus (4.6%), Citrobacter (3.7%), Actinobacillus (3.7%),
Escherichia (4.6%), Corynebacterium (4.6%) and others (8.2%). It was interesting to notice that most of the
isolates were Gram-negative bacillus (63.3%) and few were Gram-positive cocci (36.7%). Pathogenic
microorganisms from domestic wastes and tannery effluents have been identified and reported. Most of the
isolates were resistant to chloramphenicol, nalidixic acid, nitrofurontoin and cefixime. Levofloxacin and
imipenem were effective against 108 (99.5%) of the isolates. Multi drug resistance was observed in most of the
isolates. Some isolates were found in both domestic and tannery waste samples, but their antibiotic resistance
patterns were not similar. Serratia spp. and two Bacillus spp. with different antibiogram pattern were found only in tannery waste samples. The significant number of Multiple Antibiotic Resistant (MAR) bacteria was observed in both the samples. Human infections caused by these bacteria could be difficult to treat with available drugs.
Dissertation ppt biostimulation- a potential practice for wastewater treaat...Sumer Pankaj
Phycoremediation is a green technology that supports the direct use of living green microalgae for in situ, or in place removal, degradation, of contaminants in soils, sludge, sediments, surface water and ground waters by the mechanisms of bio-transformation, bio-accumulation, bio-concentration, bio-sparging.
It can be said by the current study that microalgae has a great potential for the treatment of industrial and municipal wastewaters as compared to the chemical treatments available commercially. Biological systems are much more efficient in cleaning the excess nutrients from the waste water followed by generation of valuable biomass which can be applied in the food, fertilizer, energy production as use of inorganic chemicals like lime and ferrous sulphate generates huge amount of sludge in textile industries, but on the other hand static anaerobic treatment using acclimatized MLSS gives better colour reduction with zero sludge generation. Microalgal cells can be used in free form to treat waste waters containing high C.O.D., high ammonical nitrogen and high TDS. It not only provides a better reduction of chemicals from wastewaters but it also helps to reduce the operational cost of ETP. Microalgaes not only helps to remediate industrial waste waters but also to treat sweage water and to restore natural water bodies like lakes and ponds. As they are active in remediating the chemicals but also it shows an antagonistic effect against some pathogenic germs like total coliforms and fecal coliforms.
These microalgal cells can also be combined with bacterial biomass of activated sludge process to develop an Algal-Bacterial consortium (ALBA) for better enhancement in the reduction of chemicals from the wastewaters as this symbiotic relation of algae and bacteria provides high satiability of the microalgae along with MLSS and faceable in terms of price and economy for instance the bacterial biomass provides carbon dioxide to algal cells for photosynthesis and in return the bacteria acquires oxygen from algae. The harvested biomass from the ETP’s can be used as bio-fertilizers as it consists of appropriate ratio of vital macro and micro nutrients like N,P,K etc. which enhance the growth of plantlets. It can also be used as aqua feeds for shrimps, fishes and molluscs. Furthermore these microlgal cells are non-toxic in the environment as it becomes a part of food chain and do not cause eutrophication. Therefore, micro-algal based treatment is most suitable for the treating the waste waters and restoring the natural water bodies as compared to other chemical treatments.
Isolation, characterization of aspergillus fumigatus and optimization of cult...eSAT Publishing House
IJRET : International Journal of Research in Engineering and Technology is an international peer reviewed, online journal published by eSAT Publishing House for the enhancement of research in various disciplines of Engineering and Technology. The aim and scope of the journal is to provide an academic medium and an important reference for the advancement and dissemination of research results that support high-level learning, teaching and research in the fields of Engineering and Technology. We bring together Scientists, Academician, Field Engineers, Scholars and Students of related fields of Engineering and Technology
Isolation, characterization of aspergillus fumigatus and optimization of cult...eSAT Journals
Abstract The soil samples were collected from different depths of paddy and sugarcane fields. The samples were primarily screened for isolation of amylase producing fungi. Among the isolated fungi, amylase producing isolates were identified by growing on starch agar media. The isolate (15F) which form the maximum zone of clearance on starch agar media by iodine was identified and it was subcultured on potato dextrose agar (PDA). The isolate (15F) was morphologically characterized by performing cotton blue staining and scanning electron microscopic observations under required magnifications. Molecular characterization of isolate (15F) was performed by ITS/5.8S rRNA and β-tubulin gene sequence analysis and it was confirmed as Aspergillus fumigatus (MTCC Acc No 11399). Optimization of cultural conditions for maximum production of amylase was carried out by different cereal flours, incubation periods, temperatures, nitrogen sources and with different phosphate concentrations. Aspergillus fumigatus showed maximum amylase activity (230±0.7U/mg protein) when cultured in finger millet at 350C for 72hrs of incubation period. Keywords: Amylase, Aspergillus fumigatus, cereal flour, submerged fermentation
Similar to Isolation and screening of haloalkaline protease producing bacteria from tannery solid waste (20)
Mechanical properties of hybrid fiber reinforced concrete for pavementseSAT Journals
Abstract
The effect of addition of mono fibers and hybrid fibers on the mechanical properties of concrete mixture is studied in the present
investigation. Steel fibers of 1% and polypropylene fibers 0.036% were added individually to the concrete mixture as mono fibers and
then they were added together to form a hybrid fiber reinforced concrete. Mechanical properties such as compressive, split tensile and
flexural strength were determined. The results show that hybrid fibers improve the compressive strength marginally as compared to
mono fibers. Whereas, hybridization improves split tensile strength and flexural strength noticeably.
Keywords:-Hybridization, mono fibers, steel fiber, polypropylene fiber, Improvement in mechanical properties.
Material management in construction – a case studyeSAT Journals
Abstract
The objective of the present study is to understand about all the problems occurring in the company because of improper application
of material management. In construction project operation, often there is a project cost variance in terms of the material, equipments,
manpower, subcontractor, overhead cost, and general condition. Material is the main component in construction projects. Therefore,
if the material management is not properly managed it will create a project cost variance. Project cost can be controlled by taking
corrective actions towards the cost variance. Therefore a methodology is used to diagnose and evaluate the procurement process
involved in material management and launch a continuous improvement was developed and applied. A thorough study was carried
out along with study of cases, surveys and interviews to professionals involved in this area. As a result, a methodology for diagnosis
and improvement was proposed and tested in selected projects. The results obtained show that the main problem of procurement is
related to schedule delays and lack of specified quality for the project. To prevent this situation it is often necessary to dedicate
important resources like money, personnel, time, etc. To monitor and control the process. A great potential for improvement was
detected if state of the art technologies such as, electronic mail, electronic data interchange (EDI), and analysis were applied to the
procurement process. These helped to eliminate the root causes for many types of problems that were detected.
Managing drought short term strategies in semi arid regions a case studyeSAT Journals
Abstract
Drought management needs multidisciplinary action. Interdisciplinary efforts among the experts in various fields of the droughts
prone areas are helpful to achieve tangible and permanent solution for this recurring problem. The Gulbarga district having the total
area around 16, 240 sq.km, and accounts 8.45 per cent of the Karnataka state area. The district has been situated with latitude 17º 19'
60" North and longitude of 76 º 49' 60" east. The district is situated entirely on the Deccan plateau positioned at a height of 300 to
750 m above MSL. Sub-tropical, semi-arid type is one among the drought prone districts of Karnataka State. The drought
management is very important for a district like Gulbarga. In this paper various short term strategies are discussed to mitigate the
drought condition in the district.
Keywords: Drought, South-West monsoon, Semi-Arid, Rainfall, Strategies etc.
Life cycle cost analysis of overlay for an urban road in bangaloreeSAT Journals
Abstract
Pavements are subjected to severe condition of stresses and weathering effects from the day they are constructed and opened to traffic
mainly due to its fatigue behavior and environmental effects. Therefore, pavement rehabilitation is one of the most important
components of entire road systems. This paper highlights the design of concrete pavement with added mono fibers like polypropylene,
steel and hybrid fibres for a widened portion of existing concrete pavement and various overlay alternatives for an existing
bituminous pavement in an urban road in Bangalore. Along with this, Life cycle cost analyses at these sections are done by Net
Present Value (NPV) method to identify the most feasible option. The results show that though the initial cost of construction of
concrete overlay is high, over a period of time it prove to be better than the bituminous overlay considering the whole life cycle cost.
The economic analysis also indicates that, out of the three fibre options, hybrid reinforced concrete would be economical without
compromising the performance of the pavement.
Keywords: - Fatigue, Life cycle cost analysis, Net Present Value method, Overlay, Rehabilitation
Laboratory studies of dense bituminous mixes ii with reclaimed asphalt materialseSAT Journals
Abstract
The issue of growing demand on our nation’s roadways over that past couple of decades, decreasing budgetary funds, and the need to
provide a safe, efficient, and cost effective roadway system has led to a dramatic increase in the need to rehabilitate our existing
pavements and the issue of building sustainable road infrastructure in India. With these emergency of the mentioned needs and this
are today’s burning issue and has become the purpose of the study.
In the present study, the samples of existing bituminous layer materials were collected from NH-48(Devahalli to Hassan) site.The
mixtures were designed by Marshall Method as per Asphalt institute (MS-II) at 20% and 30% Reclaimed Asphalt Pavement (RAP).
RAP material was blended with virgin aggregate such that all specimens tested for the, Dense Bituminous Macadam-II (DBM-II)
gradation as per Ministry of Roads, Transport, and Highways (MoRT&H) and cost analysis were carried out to know the economics.
Laboratory results and analysis showed the use of recycled materials showed significant variability in Marshall Stability, and the
variability increased with the increase in RAP content. The saving can be realized from utilization of recycled materials as per the
methodology, the reduction in the total cost is 19%, 30%, comparing with the virgin mixes.
Keywords: Reclaimed Asphalt Pavement, Marshall Stability, MS-II, Dense Bituminous Macadam-II
Laboratory investigation of expansive soil stabilized with natural inorganic ...eSAT Journals
Abstract
Soil stabilization has proven to be one of the oldest techniques to improve the soil properties. Literature review conducted revealed
that uses of natural inorganic stabilizers are found to be one of the best options for soil stabilization. In this regard an attempt has
been made to evaluate the influence of RBI-81 stabilizer on properties of black cotton soil through laboratory investigations. Black
cotton soil with varying percentages of RBI-81 viz., 0, 0.5, 1, 1.5, 2, and 2.5 percent were studied for moisture density relationships
and strength behaviour of soils. Also the effect of curing period was evaluated as literature review clearly emphasized the strength
gain of soils stabilized with RBI-81 over a period of time. The results obtained shows that the unconfined compressive strength of
specimens treated with RBI-81 increased approximately by 250% for a curing period of 28 days as compared to virgin soil. Further
the CBR value improved approximately by 400%. The studies indicated an increasing trend for soil strength behaviour with
increasing percentage of RBI-81 suggesting its potential applications in soil stabilization.
Influence of reinforcement on the behavior of hollow concrete block masonry p...eSAT Journals
Abstract
Reinforced masonry was developed to exploit the strength potential of masonry and to solve its lack of tensile strength. Experimental
and analytical studies have been carried out to investigate the effect of reinforcement on the behavior of hollow concrete block
masonry prisms under compression and to predict ultimate failure compressive strength. In the numerical program, three dimensional
non-linear finite elements (FE) model based on the micro-modeling approach is developed for both unreinforced and reinforced
masonry prisms using ANSYS (14.5). The proposed FE model uses multi-linear stress-strain relationships to model the non-linear
behavior of hollow concrete block, mortar, and grout. Willam-Warnke’s five parameter failure theory has been adopted to model the
failure of masonry materials. The comparison of the numerical and experimental results indicates that the FE models can successfully
capture the highly nonlinear behavior of the physical specimens and accurately predict their strength and failure mechanisms.
Keywords: Structural masonry, Hollow concrete block prism, grout, Compression failure, Finite element method,
Numerical modeling.
Influence of compaction energy on soil stabilized with chemical stabilizereSAT Journals
Abstract
Increase in traffic along with heavier magnitude of wheel loads cause rapid deterioration in pavements. There is a need to improve
density, strength of soil subgrade and other pavement layers. In this study an attempt is made to improve the properties of locally
available loamy soil using twin approaches viz., i) increasing the compaction of soil and ii) treating the soil with chemical stabilizer.
Laboratory studies are carried out on both untreated and treated soil samples compacted by different compaction efforts. Studies
show that increase in compaction effort results in increase in density of soil. However in soil treated with chemical stabilizer, rate of
increase in density is not significant. The soil treated with chemical stabilizer exhibits improvement in both strength and performance
properties.
Keywords: compaction, density, subgradestabilization, resilient modulus
Geographical information system (gis) for water resources managementeSAT Journals
Abstract
Water resources projects are inherited with overlapping and at times conflicting objectives. These projects are often of varied sizes
ranging from major projects with command areas of millions of hectares to very small projects implemented at the local level. Thus,
in all these projects there is seldom proper coordination which is essential for ensuring collective sustainability.
Integrated watershed development and management is the accepted answer but in turn requires a comprehensive framework that can
enable planning process involving all the stakeholders at different levels and scales is compulsory. Such a unified hydrological
framework is essential to evaluate the cause and effect of all the proposed actions within the drainage basins.
The present paper describes a hydrological framework developed in the form of a Hydrologic Information System (HIS) which is
intended to meet the specific information needs of the various line departments of a typical State connected with water related aspects.
The HIS consist of a hydrologic information database coupled with tools for collating primary and secondary data and tools for
analyzing and visualizing the data and information. The HIS also incorporates hydrological model base for indirect assessment of
various entities of water balance in space and time. The framework would be maintained and updated to reflect fully the most
accurate ground truth data and the infrastructure requirements for planning and management.
Keywords: Hydrological Information System (HIS); WebGIS; Data Model; Web Mapping Services
Forest type mapping of bidar forest division, karnataka using geoinformatics ...eSAT Journals
Abstract
The study demonstrate the potentiality of satellite remote sensing technique for the generation of baseline information on forest types
including tree plantation details in Bidar forest division, Karnataka covering an area of 5814.60Sq.Kms. The Total Area of Bidar
forest division is 5814Sq.Kms analysis of the satellite data in the study area reveals that about 84% of the total area is Covered by
crop land, 1.778% of the area is covered by dry deciduous forest, 1.38 % of mixed plantation, which is very threatening to the
environmental stability of the forest, future plantation site has been mapped. With the use of latest Geo-informatics technology proper
and exact condition of the trees can be observed and necessary precautions can be taken for future plantation works in an appropriate
manner
Keywords:-RS, GIS, GPS, Forest Type, Tree Plantation
Factors influencing compressive strength of geopolymer concreteeSAT Journals
Abstract
To study effects of several factors on the properties of fly ash based geopolymer concrete on the compressive strength and also the
cost comparison with the normal concrete. The test variables were molarities of sodium hydroxide(NaOH) 8M,14M and 16M, ratio of
NaOH to sodium silicate (Na2SiO3) 1, 1.5, 2 and 2.5, alkaline liquid to fly ash ratio 0.35 and 0.40 and replacement of water in
Na2SiO3 solution by 10%, 20% and 30% were used in the present study. The test results indicated that the highest compressive
strength 54 MPa was observed for 16M of NaOH, ratio of NaOH to Na2SiO3 2.5 and alkaline liquid to fly ash ratio of 0.35. Lowest
compressive strength of 27 MPa was observed for 8M of NaOH, ratio of NaOH to Na2SiO3 is 1 and alkaline liquid to fly ash ratio of
0.40. Alkaline liquid to fly ash ratio of 0.35, water replacement of 10% and 30% for 8 and 16 molarity of NaOH and has resulted in
compressive strength of 36 MPa and 20 MPa respectively. Superplasticiser dosage of 2 % by weight of fly ash has given higher
strength in all cases.
Keywords: compressive strength, alkaline liquid, fly ash
Experimental investigation on circular hollow steel columns in filled with li...eSAT Journals
Abstract
Composite Circular hollow Steel tubes with and without GFRP infill for three different grades of Light weight concrete are tested for
ultimate load capacity and axial shortening , under Cyclic loading. Steel tubes are compared for different lengths, cross sections and
thickness. Specimens were tested separately after adopting Taguchi’s L9 (Latin Squares) Orthogonal array in order to save the initial
experimental cost on number of specimens and experimental duration. Analysis was carried out using ANN (Artificial Neural
Network) technique with the assistance of Mini Tab- a statistical soft tool. Comparison for predicted, experimental & ANN output is
obtained from linear regression plots. From this research study, it can be concluded that *Cross sectional area of steel tube has most
significant effect on ultimate load carrying capacity, *as length of steel tube increased- load carrying capacity decreased & *ANN
modeling predicted acceptable results. Thus ANN tool can be utilized for predicting ultimate load carrying capacity for composite
columns.
Keywords: Light weight concrete, GFRP, Artificial Neural Network, Linear Regression, Back propagation, orthogonal
Array, Latin Squares
Experimental behavior of circular hsscfrc filled steel tubular columns under ...eSAT Journals
Abstract
This paper presents an outlook on experimental behavior and a comparison with predicted formula on the behaviour of circular
concentrically loaded self-consolidating fibre reinforced concrete filled steel tube columns (HSSCFRC). Forty-five specimens were
tested. The main parameters varied in the tests are: (1) percentage of fiber (2) tube diameter or width to wall thickness ratio (D/t
from 15 to 25) (3) L/d ratio from 2.97 to 7.04 the results from these predictions were compared with the experimental data. The
experimental results) were also validated in this study.
Keywords: Self-compacting concrete; Concrete-filled steel tube; axial load behavior; Ultimate capacity.
Evaluation of punching shear in flat slabseSAT Journals
Abstract
Flat-slab construction has been widely used in construction today because of many advantages that it offers. The basic philosophy in
the design of flat slab is to consider only gravity forces; this method ignores the effect of punching shear due to unbalanced moments
at the slab column junction which is critical. An attempt has been made to generate generalized design sheets which accounts both
punching shear due to gravity loads and unbalanced moments for cases (a) interior column; (b) edge column (bending perpendicular
to shorter edge); (c) edge column (bending parallel to shorter edge); (d) corner column. These design sheets are prepared as per
codal provisions of IS 456-2000. These design sheets will be helpful in calculating the shear reinforcement to be provided at the
critical section which is ignored in many design offices. Apart from its usefulness in evaluating punching shear and the necessary
shear reinforcement, the design sheets developed will enable the designer to fix the depth of flat slab during the initial phase of the
design.
Keywords: Flat slabs, punching shear, unbalanced moment.
Evaluation of performance of intake tower dam for recent earthquake in indiaeSAT Journals
Abstract
Intake towers are typically tall, hollow, reinforced concrete structures and form entrance to reservoir outlet works. A parametric
study on dynamic behavior of circular cylindrical towers can be carried out to study the effect of depth of submergence, wall thickness
and slenderness ratio, and also effect on tower considering dynamic analysis for time history function of different soil condition and
by Goyal and Chopra accounting interaction effects of added hydrodynamic mass of surrounding and inside water in intake tower of
dam
Key words: Hydrodynamic mass, Depth of submergence, Reservoir, Time history analysis,
Evaluation of operational efficiency of urban road network using travel time ...eSAT Journals
Abstract
Efficiency of the road network system is analyzed by travel time reliability measures. The study overlooks on an important measure of
travel time reliability and prioritizing Tiruchirappalli road network. Traffic volume and travel time were collected using license plate
matching method. Travel time measures were estimated from average travel time and 95th travel time. Effect of non-motorized vehicle
on efficiency of road system was evaluated. Relation between buffer time index and traffic volume was created. Travel time model has
been developed and travel time measure was validated. Then service quality of road sections in network were graded based on
travel time reliability measures.
Keywords: Buffer Time Index (BTI); Average Travel Time (ATT); Travel Time Reliability (TTR); Buffer Time (BT).
Estimation of surface runoff in nallur amanikere watershed using scs cn methodeSAT Journals
Abstract
The development of watershed aims at productive utilization of all the available natural resources in the entire area extending from
ridge line to stream outlet. The per capita availability of land for cultivation has been decreasing over the years. Therefore, water and
the related land resources must be developed, utilized and managed in an integrated and comprehensive manner. Remote sensing and
GIS techniques are being increasingly used for planning, management and development of natural resources. The study area, Nallur
Amanikere watershed geographically lies between 110 38’ and 110 52’ N latitude and 760 30’ and 760 50’ E longitude with an area of
415.68 Sq. km. The thematic layers such as land use/land cover and soil maps were derived from remotely sensed data and overlayed
through ArcGIS software to assign the curve number on polygon wise. The daily rainfall data of six rain gauge stations in and around
the watershed (2001-2011) was used to estimate the daily runoff from the watershed using Soil Conservation Service - Curve Number
(SCS-CN) method. The runoff estimated from the SCS-CN model was then used to know the variation of runoff potential with different
land use/land cover and with different soil conditions.
Keywords: Watershed, Nallur watershed, Surface runoff, Rainfall-Runoff, SCS-CN, Remote Sensing, GIS.
Estimation of morphometric parameters and runoff using rs & gis techniqueseSAT Journals
Abstract
Land and water are the two vital natural resources, the optimal management of these resources with minimum adverse environmental
impact are essential not only for sustainable development but also for human survival. Satellite remote sensing with geographic
information system has a pragmatic approach to map and generate spatial input layers of predicting response behavior and yield of
watershed. Hence, in the present study an attempt has been made to understand the hydrological process of the catchment at the
watershed level by drawing the inferences from moprhometric analysis and runoff. The study area chosen for the present study is
Yagachi catchment situated in Chickamaglur and Hassan district lies geographically at a longitude 75⁰52’08.77”E and
13⁰10’50.77”N latitude. It covers an area of 559.493 Sq.km. Morphometric analysis is carried out to estimate morphometric
parameters at Micro-watershed to understand the hydrological response of the catchment at the Micro-watershed level. Daily runoff
is estimated using USDA SCS curve number model for a period of 10 years from 2001 to 2010. The rainfall runoff relationship of the
study shows there is a positive correlation.
Keywords: morphometric analysis, runoff, remote sensing and GIS, SCS - method
-
Effect of variation of plastic hinge length on the results of non linear anal...eSAT Journals
Abstract The nonlinear Static procedure also well known as pushover analysis is method where in monotonically increasing loads are applied to the structure till the structure is unable to resist any further load. It is a popular tool for seismic performance evaluation of existing and new structures. In literature lot of research has been carried out on conventional pushover analysis and after knowing deficiency efforts have been made to improve it. But actual test results to verify the analytically obtained pushover results are rarely available. It has been found that some amount of variation is always expected to exist in seismic demand prediction of pushover analysis. Initial study is carried out by considering user defined hinge properties and default hinge length. Attempt is being made to assess the variation of pushover analysis results by considering user defined hinge properties and various hinge length formulations available in literature and results compared with experimentally obtained results based on test carried out on a G+2 storied RCC framed structure. For the present study two geometric models viz bare frame and rigid frame model is considered and it is found that the results of pushover analysis are very sensitive to geometric model and hinge length adopted. Keywords: Pushover analysis, Base shear, Displacement, hinge length, moment curvature analysis
Effect of use of recycled materials on indirect tensile strength of asphalt c...eSAT Journals
Abstract
Depletion of natural resources and aggregate quarries for the road construction is a serious problem to procure materials. Hence
recycling or reuse of material is beneficial. On emphasizing development in sustainable construction in the present era, recycling of
asphalt pavements is one of the effective and proven rehabilitation processes. For the laboratory investigations reclaimed asphalt
pavement (RAP) from NH-4 and crumb rubber modified binder (CRMB-55) was used. Foundry waste was used as a replacement to
conventional filler. Laboratory tests were conducted on asphalt concrete mixes with 30, 40, 50, and 60 percent replacement with RAP.
These test results were compared with conventional mixes and asphalt concrete mixes with complete binder extracted RAP
aggregates. Mix design was carried out by Marshall Method. The Marshall Tests indicated highest stability values for asphalt
concrete (AC) mixes with 60% RAP. The optimum binder content (OBC) decreased with increased in RAP in AC mixes. The Indirect
Tensile Strength (ITS) for AC mixes with RAP also was found to be higher when compared to conventional AC mixes at 300C.
Keywords: Reclaimed asphalt pavement, Foundry waste, Recycling, Marshall Stability, Indirect tensile strength.
Cosmetic shop management system project report.pdfKamal Acharya
Buying new cosmetic products is difficult. It can even be scary for those who have sensitive skin and are prone to skin trouble. The information needed to alleviate this problem is on the back of each product, but it's thought to interpret those ingredient lists unless you have a background in chemistry.
Instead of buying and hoping for the best, we can use data science to help us predict which products may be good fits for us. It includes various function programs to do the above mentioned tasks.
Data file handling has been effectively used in the program.
The automated cosmetic shop management system should deal with the automation of general workflow and administration process of the shop. The main processes of the system focus on customer's request where the system is able to search the most appropriate products and deliver it to the customers. It should help the employees to quickly identify the list of cosmetic product that have reached the minimum quantity and also keep a track of expired date for each cosmetic product. It should help the employees to find the rack number in which the product is placed.It is also Faster and more efficient way.
CW RADAR, FMCW RADAR, FMCW ALTIMETER, AND THEIR PARAMETERSveerababupersonal22
It consists of cw radar and fmcw radar ,range measurement,if amplifier and fmcw altimeterThe CW radar operates using continuous wave transmission, while the FMCW radar employs frequency-modulated continuous wave technology. Range measurement is a crucial aspect of radar systems, providing information about the distance to a target. The IF amplifier plays a key role in signal processing, amplifying intermediate frequency signals for further analysis. The FMCW altimeter utilizes frequency-modulated continuous wave technology to accurately measure altitude above a reference point.
Immunizing Image Classifiers Against Localized Adversary Attacksgerogepatton
This paper addresses the vulnerability of deep learning models, particularly convolutional neural networks
(CNN)s, to adversarial attacks and presents a proactive training technique designed to counter them. We
introduce a novel volumization algorithm, which transforms 2D images into 3D volumetric representations.
When combined with 3D convolution and deep curriculum learning optimization (CLO), itsignificantly improves
the immunity of models against localized universal attacks by up to 40%. We evaluate our proposed approach
using contemporary CNN architectures and the modified Canadian Institute for Advanced Research (CIFAR-10
and CIFAR-100) and ImageNet Large Scale Visual Recognition Challenge (ILSVRC12) datasets, showcasing
accuracy improvements over previous techniques. The results indicate that the combination of the volumetric
input and curriculum learning holds significant promise for mitigating adversarial attacks without necessitating
adversary training.
Using recycled concrete aggregates (RCA) for pavements is crucial to achieving sustainability. Implementing RCA for new pavement can minimize carbon footprint, conserve natural resources, reduce harmful emissions, and lower life cycle costs. Compared to natural aggregate (NA), RCA pavement has fewer comprehensive studies and sustainability assessments.
Overview of the fundamental roles in Hydropower generation and the components involved in wider Electrical Engineering.
This paper presents the design and construction of hydroelectric dams from the hydrologist’s survey of the valley before construction, all aspects and involved disciplines, fluid dynamics, structural engineering, generation and mains frequency regulation to the very transmission of power through the network in the United Kingdom.
Author: Robbie Edward Sayers
Collaborators and co editors: Charlie Sims and Connor Healey.
(C) 2024 Robbie E. Sayers
Saudi Arabia stands as a titan in the global energy landscape, renowned for its abundant oil and gas resources. It's the largest exporter of petroleum and holds some of the world's most significant reserves. Let's delve into the top 10 oil and gas projects shaping Saudi Arabia's energy future in 2024.
Hierarchical Digital Twin of a Naval Power SystemKerry Sado
A hierarchical digital twin of a Naval DC power system has been developed and experimentally verified. Similar to other state-of-the-art digital twins, this technology creates a digital replica of the physical system executed in real-time or faster, which can modify hardware controls. However, its advantage stems from distributing computational efforts by utilizing a hierarchical structure composed of lower-level digital twin blocks and a higher-level system digital twin. Each digital twin block is associated with a physical subsystem of the hardware and communicates with a singular system digital twin, which creates a system-level response. By extracting information from each level of the hierarchy, power system controls of the hardware were reconfigured autonomously. This hierarchical digital twin development offers several advantages over other digital twins, particularly in the field of naval power systems. The hierarchical structure allows for greater computational efficiency and scalability while the ability to autonomously reconfigure hardware controls offers increased flexibility and responsiveness. The hierarchical decomposition and models utilized were well aligned with the physical twin, as indicated by the maximum deviations between the developed digital twin hierarchy and the hardware.
NUMERICAL SIMULATIONS OF HEAT AND MASS TRANSFER IN CONDENSING HEAT EXCHANGERS...ssuser7dcef0
Power plants release a large amount of water vapor into the
atmosphere through the stack. The flue gas can be a potential
source for obtaining much needed cooling water for a power
plant. If a power plant could recover and reuse a portion of this
moisture, it could reduce its total cooling water intake
requirement. One of the most practical way to recover water
from flue gas is to use a condensing heat exchanger. The power
plant could also recover latent heat due to condensation as well
as sensible heat due to lowering the flue gas exit temperature.
Additionally, harmful acids released from the stack can be
reduced in a condensing heat exchanger by acid condensation. reduced in a condensing heat exchanger by acid condensation.
Condensation of vapors in flue gas is a complicated
phenomenon since heat and mass transfer of water vapor and
various acids simultaneously occur in the presence of noncondensable
gases such as nitrogen and oxygen. Design of a
condenser depends on the knowledge and understanding of the
heat and mass transfer processes. A computer program for
numerical simulations of water (H2O) and sulfuric acid (H2SO4)
condensation in a flue gas condensing heat exchanger was
developed using MATLAB. Governing equations based on
mass and energy balances for the system were derived to
predict variables such as flue gas exit temperature, cooling
water outlet temperature, mole fraction and condensation rates
of water and sulfuric acid vapors. The equations were solved
using an iterative solution technique with calculations of heat
and mass transfer coefficients and physical properties.
Sachpazis:Terzaghi Bearing Capacity Estimation in simple terms with Calculati...Dr.Costas Sachpazis
Terzaghi's soil bearing capacity theory, developed by Karl Terzaghi, is a fundamental principle in geotechnical engineering used to determine the bearing capacity of shallow foundations. This theory provides a method to calculate the ultimate bearing capacity of soil, which is the maximum load per unit area that the soil can support without undergoing shear failure. The Calculation HTML Code included.
Student information management system project report ii.pdfKamal Acharya
Our project explains about the student management. This project mainly explains the various actions related to student details. This project shows some ease in adding, editing and deleting the student details. It also provides a less time consuming process for viewing, adding, editing and deleting the marks of the students.
HEAP SORT ILLUSTRATED WITH HEAPIFY, BUILD HEAP FOR DYNAMIC ARRAYS.
Heap sort is a comparison-based sorting technique based on Binary Heap data structure. It is similar to the selection sort where we first find the minimum element and place the minimum element at the beginning. Repeat the same process for the remaining elements.
Isolation and screening of haloalkaline protease producing bacteria from tannery solid waste
1. IJRET: International Journal of Research in Engineering and Technology eISSN: 2319-1163 | pISSN: 2321-7308
_______________________________________________________________________________________
Volume: 05 Issue: 01 | Jan-2016, Available @ http://www.ijret.org 237
ISOLATION AND SCREENING OF HALOALKALINE PROTEASE
PRODUCING BACTERIA FROM TANNERY SOLID WASTE
Tuhina Verma1
, Swati Agarwal2
1
Assistant Professor, Department of Microbiology (Centre of Excellence), Dr. Ram Manohar Lohia Avadh University,
Faizabad, Uttar Pradesh, India
2
Research Scholar, Department of Microbiology (Centre of Excellence), Dr. Ram Manohar Lohia Avadh University,
Faizabad, Uttar Pradesh, India
*1
Corresponding author: tuhinaverma19@gmail.com
Abstract
Twenty bacterial strains were isolated on selective milk agar plates (pH 9.0) from tannery solid waste on the basis of different
colony morphology. These strains exhibited variable haloalkaline protease activity and were tolerant to different concentration of
both chromate (350-1450 μg/ml) and NaCl (2-9%). Those having clearance zone greater than 20.0 mm were considered as
significant isolate. Out of twenty, nine strains were interestingly tolerant to high concentration of Cr(VI) (850-1450 mg/l) and
NaCl (6.5-9%) and exhibited vibrant clear zone diameter between 21-35 mm. All these isolates in this study were capable of
reducing Cr(VI) aerobically and the reduction values ranged between 50.0-76.0% after 16-20 h of growth. Further, significant
haloalkaline protease production was observed in these bacteria within 24 h under wide temperature (25-45°C) and pH (8.0-10.5)
range. The potential of these strains to produce protease at higher rate in the presence of wheat bran as a cheap carbon source
and yeast extract or beef extract as a nitrogen source makes them a potential candidate for industrial applications and removal of
Cr(VI) and proteinaceous waste simultaneously from industrial waste of alkaline nature.
Key words: Tannery waste, chromate resistant, bacteria, haloalkaline, protease
--------------------------------------------------------------------***----------------------------------------------------------------------
1. INTRODUCTION
Protease, a hydrolytic enzyme has attracted much attention
because of its wide application in detergent, leather, food,
pharmaceutical, agricultural industries and proteinaceous
waste bioremediation. Of all industrially important enzymes,
proteases constitute >65% of the total enzymes employed in
various industrial and commercial purposes [1]. Among the
various type of proteases, bacterial alkaline proteases are the
most significant and widely studied group of enzymes
compared with animal, plant and fungal proteases as they
exhibit high productivity rate, cheaper production cost,
stability to chemical and physical changes in the medium
and are more demanding for industries [2, 3]. Interestingly,
extracellular haloalkaline bacterial proteases are important
for the hydrolysis of waste proteins with dual extremities of
high salt and alkaline pH and enable the bacteria to absorb
and utilize hydrolytic products under harsh conditions [4].
Recently, such proteases have received attention as a viable
alternative for bioremediation of protein rich tannery waste
and their use in treatment of raw hide by replacing the
hazardous chemicals especially involved in soaking,
dehairing and bating of hides prior to tanning to produce
quality leather without causing environmental pollution [5].
Further, many industrial processes are carried out in saline
environments, where most of the normal bacterial alkaline
proteases becomes unstable [6, 7]. Moreover, there is very
limited information on the salt tolerant extracellular alkaline
protease producing bacteria from the natural environment.
Leather industries are one of the major industrial sectors of
India but are recognized as one of the highly polluting
industries leading to soil and water pollution [8, 9].
Tanneries are mainly responsible for the release of huge
amount of proteinaceous substances and toxic hexavalent
chromium [Cr (VI), chromate] through their solid waste into
the environment and hence are of great environmental
concern. Protein degradation emits obnoxious smell due to
generation of toxic gases such as NH3, H2S and CO2 that
creates serious human health problem. Hexavalent
chromium [Cr (VI)] is well known carcinogen, mutagen and
teratogen and cause cellular toxicity in humans and other
living beings, thus listed as priority pollutant by
Environmental Protection Agency [10]. Also, the tanneries
use sodium chloride to preserve the fresh skins from
decomposition, thus the tannery wastes are saline in nature
having basic pH.
Bacteria inhabiting in such environments are a remarkable
source for producing halotolerant alkaline proteases which
will have diverse potential applications in the industries
including environmental restoration [3]. In a study,
Sivaprakasam et al. [4] have reported the production of a
salt tolerant protease from Pseudomonas aeruginosa BC1
and its application in tannery saline wastewater treatment.
Therefore, conducting research for the isolation of
haloalkalophilic bacteria capable to produce extracellular
proteases are of great importance. Also, these bacteria could
be utilized for removal of both proteinaceous waste and Cr
(VI) from industrial wastes. Keeping it in view, this study
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was aimed to screen alkaline protease producing bacteria
from the discharged tannery solid waste as it may result in
isolation of novel haloalkaline proteases with unique
physico-chemical characteristics with a view to facilitate its
potential application in industries.
2. MATERIALS AND METHODS
2.1. Media and Chemicals
The media components and all reagents used in the study
were of analytical grade and purchased from Hi-Media,
Merck, Qualigens, India Ltd. and Sigma Aldrich chemicals,
USA. Diphenylcarbazide (DPC) solution (0.25%, w/v) was
prepared by dissolving 125 mg 1,5- diphenylcarbazide in 25
ml of high performance liquid chromatography (HPLC)
grade acetone and stored in a brown glass bottle, for Cr(VI)
determination.
2.2. Sampling and isolation of chromate resistant
haloalkaliphilic protease producing bacteria
The tannery solid waste was collected from the dumping
sites of the tanneries of Jajmau, Kanpur, India and
landfilling sites of vicinity area in sterile containers,
transported on ice to the laboratory and was processed for
isolation of chromate resistant haloalkaliphilic protease
producing bacteria within 6-8 h of collection. One gram of
solid waste was serially diluted with sterile distilled water
and the bacteria were isolated on the saline skim milk agar
plates containing 1.0% (w/v) skimmed milk, 0.5% (w/v)
peptone, 1.0% (w/v) NaCl and 1.5% (w/v) agar by the
standard pour plate technique [11]. The agar plates were
also supplemented with varying chromate (100-250 mg/l)
and NaCl (1.0-1.5%) concentration. The pH of the media
was adjusted to 9.0 after autoclaving with previously
sterilized Na2CO3 (20% w/v). Plates were then incubated at
35+1°C for 24-36 h. Bacterial colonies showing transparent
zones around their colony due to hydrolysis of milk casein,
after 24 h of incubation were taken as evidence for
qualitative determination of protease producing bacteria.
Twenty morphologically distinct bacterial colonies showing
the clear zone diameter greater than 12.0 mm were selected
and re-streaked several times on the same medium to obtain
pure isolates.
2.3. Screening of bacteria for tolerance to high
Cr(VI) and NaCl concentration
All the twenty bacterial isolates were screened for resistance
to high Cr(VI) and NaCl concentration by determining the
MIC for Cr(VI) by agar dilution method having varying
concentrations of NaCl. The milk agar plates supplemented
with different concentrations of Cr(VI) (200–1550 mg/l) and
NaCl (1.5–9.5%) in combination were inoculated aseptically
with about 2.9 X 105
colony forming units (CFU)/ ml
bacterial cells of exponential phase. Plates were incubated
for 24 h at 35+1°C and observed for growth. The minimum
concentration of Cr(VI), at which no growth observed, was
considered the MIC of that isolate. Also, the maximum
NaCl tolerance concentration of these strains was recorded.
Nine strains exhibiting high proteolytic activity and showing
higher MIC values for Cr(VI) and tolerance to high NaCl
concentration were selected for further study.
2.4. Assay of Cr(VI) reduction activity
Potential of selected nine strains were evaluated for the
reduction of Cr(VI) to less toxic Cr(III) at the respective
maximum chromate tolerance concentrations. Chromate
concentration was determined spectrophotometrically in
supernatant fraction of overnight grown cultures by
diphenylcarbazide (DPC) method at 540 nm using UV-Vis
spectrophotometer (Shimadzu 1601, Japan) and the Cr(VI)
concentration was determined by the standard curve of
K2Cr2O7 (100–1500 mg/l) [11]. The initial (0 h) and final
(after incubation) Cr(VI) concentration was determined by
the DPC method and the Cr(VI) reduction efficiency of
bacteria is determined in terms of “% Cr(VI) reduction”.
Total Cr [Cr(VI) + Cr(III)] in the culture supernatant was
determined by atomic absorption spectrophotometer (AAS)
at 357.9 nm, after digesting the supernatant with the mixture
of nitric acid and perchloric acid (6:1, v/v).
2.5. Preperation of crude extracellular alkaline
protease extract and enzyme assay
The crude extracellular alkaline protease of selected bacteria
was prepared by inoculating each strain individually into 50
ml of sterilized skim milk broth (pH 9.0) in 150 ml
Erlenmeyer conical flask and incubated at 35+1°C up to 30
h in an orbital shaker (120 rpm). The samples were
withdrawn aseptically after regular interval of every 4 h up
to 30 h of growth. Each sample was centrifuged at 10,000
rpm and 4°C for 5 min and the cell-free supernatant were
collected and used as a crude enzyme extract for
extracellular protease assay.
Enzyme activity was assayed using casein as the substrate
with slight modification to the method of [12]. The reaction
mixture consisted of 0.25 ml of 50 mM sodium phosphate
buffer (pH 7.0) containing 2.0% (w/v) of casein and 0.15 ml
of enzyme solution. The reaction mixture was incubated at
25ºC for 15 min thereafter stopped by adding 1.2 ml of
10.0% (w/v) TCA then incubated at 37ºC for an additional
15 min, and the precipitate was removed by centrifugation at
8,000 rpm for 5 min. Further 1.4 ml of 1.0 M NaOH was
added to 1.2 ml of the supernatant, and its absorbance was
measured at 600 nm. The activity was determined by
detecting the release of amino acids (tyrosine) from casein
and the amount of tyrosine released was calculated from the
standard curve constructed with tyrosine [13]. One unit of
protease activity is defined as the amount of enzyme
required to liberate 1.0 μg of tyrosine per min per ml under
the standard assay conditions.
2.6. Bacterial growth response
Growth response of selected strains was monitored at
definite time intervals by measuring the absorbance at 600
nm. Each strain was individually inoculated into 100 ml
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sterilized skim milk broth (pH 9.0) in 250 ml Erlenmeyer
conical flask and incubated at 35+1°C up to 30 h in an
orbital shaker (120 rpm). The samples were withdrawn
aseptically after regular interval of every 4 h up to 30 h of
growth and absorbance was recorded. Also, the protease and
Cr(VI) reduction activity of the bacteria was determined at
definite time intervals throughout the bacterial growth.
2.7. Optimization of pH and temperature for
protease production
The influence of pH on protease production was assessed by
growing bacterial culture in the skim milk broth of varying
pH ranging from 7.0 to 12.0 and the enzyme activity was
assayed as described above. Similarly, the effect of
incubation temperatures on protease production was
determined by incubating the culture broth at different
temperature viz. 15, 20, 25, 30, 35, 40, 45, 50, and 55°C
following the standard method.
2.8. Effect of carbon and nitrogen source on
protease production
The influence of various carbon and nitrogen source on
enzyme production was investigated. The bacterial growth
media was supplemented with various carbon sources such
as maltose, glucose, fructose, sucrose, lactose, starch,
glycerol and wheat bran at the level of 2.0% w/v. The effect
of various nitrogen sources such as yeast extract, beef
extract, urea, ammonium nitrate, sodium nitrate and
ammonium sulphate on protease production were also
studied. The peptone of the growth medium was replaced
with these nitrogen sources. After incubation with 1.0%
inoculum for 24 h at 35+1°C, the cell free supernatants
obtained by centrifugation at 120 rpm were assayed for the
protease production as described above.
2.9. Statistical analysis
The experiments were performed thrice, each in triplicate.
Standard deviation for each experimental result was
calculated using Microsoft Excel.
3. RESULTS AND DISCUSSION
3.1. Isolation and screening of chromate resistant
halotolerant alkaline protease producing bacteria
A total of twenty protease producing chromate resistant
haloalkaliphilic bacterial strains were isolated on selective
milk agar plates (pH 8.0-9.0) from tannery solid waste on
the basis of different colony morphology (Table 1).
Bacterial colonies showing zones of clearance around their
colony due to hydrolysis of milk casein was taken as an
evidence for qualitative protease production. These strains
exhibited variable alkaline protease activity and were
tolerant to different concentration of both chromate (400-
1450 mg/l) and NaCl (2-9%). Those having clearance zone
greater than 20.0 mm were considered as significant isolate.
Out of twenty, nine strains TVD-4, TVD-5, TVD-6, TVD-
8, TVD-9, TVD-11, TVD-12, TVD-17, and TVD-20 were
interestingly tolerant to high concentration of Cr(VI) (850-
1450 mg/l) and NaCl (6.5-9%) and exhibited vibrant clear
zone diameter between 21-35 mm on selective skim milk
agar medium at pH 9.0 after 24 h incubation and were
selected for further study. The proteolytic activity of these
nine strains were reassessed by loading their culture broth in
the wells on milk agar plate (pH 9.0) and the casein
hydrolysis zone also indicated the extracellular nature of
protease. This reveals that these bacteria were chromate
resistant and haloalkaliphilic as they grew in the presence of
high Cr(VI) and NaCl concentration and at pH 9.0. Further,
the chronic exposure to high levels of chromate, NaCl and
alkaline environment have resulted in bacterial communities
which belongs to a novel group of extremophiles that have
an exceptional ability to adapt in chromate and NaCl
polluted basic environments and have developed some
mechanisms as an effective tool for survival in such stress
environments [14].
In tanneries, such isolates may be useful for dehairing and
bating processes during tanning operations and also for
hydrolysis of proteinaceous waste in the discharged tannery
waste [2]. Several researchers have also isolated the alkaline
protease producing bacteria from tannery waste reflecting
their potential in waste water treatment and leather
manufacturing as an accepted green alternative to the
chemical process [15, 4] but the enzyme becomes unstable
when the industrial processes were carried out at dual
extremities of high pH and NaCl concentration.
Table-1: MIC of Cr(VI), NaCl tolerance efficiency and
alkaline protease production of bacteria isolated from
tannery solid waste
S.No.
Bacterial
strains
MIC of
Cr (VI)
(mg/l)
NaCl
conc.
(%)
Zone
diameter
(mm)
1 TVD-1 500 2 17
2 TVD-2 600 2.5 13
3 TVD-3 450 2 20
4 TVD-4 1150 6.5 25
5 TVD-5 1300 8 34
6 TVD-6 1200 7 27
7 TVD-7 750 4 32
8 TVD-8 1250 8 26
9 TVD-9 1500 9 45
10 TVD-10 700 5 30
11 TVD-11 900 5.5 23
12 TVD-12 1100 6.5 27
13 TVD-13 550 4 18
14 TVD-14 850 2 24
15 TVD-15 700 5 17
16 TVD-16 800 4 20
17 TVD-17 1000 5.5 23
18 TVD-18 400 2 14
19 TVD-19 650 2 19
20 TVD-20 950 6.5 21
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3.2. Growth, chromate reduction and
haloalkaliphilic protease activity of selected
bacteria
Bacterial growth of selected strains was measured at 600 nm
in selective skim milk broth and a typical sigmoidal growth
curve was obtained (Figure not shown). Onset of stationary
phase of these strains was between 16-20 h of growth and
during this phase, the protease production was found
maximum in all the nine strains (Figure 1). Production of
protease during the stationary phase of growth is in
accordance with the studies of other workers [4, 14]. In
addition to the alkaline protease production, the chromate
reduction potential of these bacteria was also determined
and the results are depicted in figure 1. All the nine bacterial
isolates in this study were capable of reducing Cr(VI)
aerobically and the reduction values ranged between 50.0-
76.0% after 16-20 h of growth. Results indicated that the
Cr(VI) reduction and NaCl tolerance ability of these strains
were growth dependent and gradually increased as the
incubation time progressed up to 16-20 h and their value
decreased in the stationary phase, whereas, the protease
activity was maximum during stationary phase and
thereafter the enzyme activity started to decline. This
correlation was attributable to an increased need for
turnover of cell proteins at the slower growth rate [16].
Further, incubation resulted into a lesser growth as well as
lesser alkaline protease production. These findings are in
agreement with the study of other researchers who reported
maximum protease activity after 24 h [17]. Other
researchers have also reported little extracellular protease
production during the lag and early log phase of the
bacterial growth, whereas, it is largely produced during the
post exponential phase or onset of stationary phase of their
growth [15, 2].
Fig-1: Growth response, Cr(VI) reduction (%) and protease activity (Unit/ml) of selected bacterial strains. Error bars represent
mean ± standard deviation. Each point is mean of three independent experiments.
3.3. Effect of temperature on protease production
Enzyme production was recorded by varying the incubation
temperature form 15-55°C. Figure 2 revealed that the
bacterial isolates yielded maximum protease production
between 25-45°C. Further, increase in temperature resulted
in the decrease of enzyme and biomass production.
Temperature was found to significantly regulate the
synthesis and secretion of bacterial extracellular protease by
changing the physical properties of the cell membrane [18,
19]. Therefore, temperature is a critical parameter that
should be considered in order to obtain maximum protease
production [20]. Further, the temperature requirement of the
organism is based on the nature of organisms. Many reports
showed bacterial and fungal alkaline protease production at
lower and moderate temperatures (20-30°C) [21, 6].
0
50
100
150
200
250
300
350
400
450
500
TVD-4 TVD-5 TVD-6 TVD-8 TVD-9 TVD-11 TVD-12 TVD-17 TVD-20
Bacterial strains
%Cr(VI)reductionand
Proteaseactivity(U/ml)
0
0.5
1
1.5
2
2.5
BacterialGrowthat600nm
% Cr (VI) reduction Protease activity Growth
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Fig-2: Effect of temperature on protease production of selected bacterial strains. Error bars represent mean ± standard deviation.
Each point is mean of three independent experiments.
3.4. Effect of pH on protease production
The influence of medium pH on protease production was
studied by varying the pH from 7.0-12.0 and a constant
respective incubation temperature for each strain showing
maximum growth. It is evident from Figure 3 that the
maximum protease production in these nine strains was
achieved at pH 8.0-10.5. Beyond this pH range, decrease in
protease production and biomass production was observed.
Further, the remarkable protease production in the pH range
of 8.0-10.5 revealed alkaliphilic nature of these strains as
well as protease enzyme. Effect of pH is one of the major
factors for the growth of microbes in the medium that
affects the productivity of microbes [22]. Also, pH of the
growth medium plays a vital role by inducing physiological
changes in microbes and their enzyme secretion [23]. It also
strongly affects enzymatic processes and transport of
compounds across the cell membrane [7]. Such proteases are
very suitable for application in several industries including
leather manufacturing. Moreover, the tannery wastewater is
alkaline in nature which favors the potential usage of the
isolated protease for bioremediation studies [24, 25].
Further, the molecular basis of pH affecting bacterial
metabolism in culture broth is obscure. Since proton motive
force in chemiosmosis is affected by the medium pH value,
it is possible that under optimum pH range, the relative
metabolic efficiency is high [26].
Fig-3: Effect of pH on protease production of selected bacterial strains. Error bars represent mean ± standard deviation. Each
point is mean of three independent experiments.
0
50
100
150
200
250
300
350
400
450
500
TVD-4 TVD-5 TVD-6 TVD-8 TVD-9 TVD-11 TVD-12 TVD-17 TVD-20
Bacterial strains
Proteaseproduction(U/ml)
0
5
10
15
20
25
30
35
40
45
50
Temperature(o
C)
Protease production (U/ml) Temperature (oC)
0
50
100
150
200
250
300
350
400
450
500
TVD-4 TVD-5 TVD-6 TVD-8 TVD-9 TVD-11 TVD-12 TVD-17 TVD-20
Bacterial strains
Proteaseproduction(U/ml)
0
2
4
6
8
10
12
pH
Protease production (U/ml) pH
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3.5. Effect of carbon and nitrogen sources on
protease production
The effect of various carbon sources on protease production
was evaluated. Table 2 revealed that out of nine bacterial
strains in seven strains (TVD-4, TVD-5, TVD-9, TVD-11,
TVD-12, TVD-17, and TVD-20) wheat bran (2.0% w/v)
was found to be the best carbon source, allowing maximum
protease production (300-450 U/ml) followed by maltose
which supported maximum protease production in TVD-6
(360 U/ml) and TVD-8 (390 U/ml) strain. The addition of
carbon sources in the form of either monosaccharides or
polysaccharides could influence the production of enzyme
[27]. However, Gaur et al. [1] reported that addition of
starch to the culture medium induced protease synthesis. In
contrast, other sugars such as glucose, fructose, maltose and
lactose reduced protease production although growth was
observed optimal [15].
Table-2: Effect of various carbon sources on protease production of selected bacteria (Unit/ml)
S.No.
Bacterial
strains
Maximum Protease production (Unit/ml) after 24 h incubation
Maltose Fructose Sucrose Lactose Starch Glycerol
Wheat
bran
1 TVD-4 340 325 305 288 270 255 360
2 TVD-5 420 435 415 390 378 355 430
3 TVD-6 300 315 268 360 265 250 340
4 TVD-8 355 390 375 382 360 330 330
5 TVD-9 410 385 425 450 430 380 450
6 TVD-11 295 310 290 245 275 230 320
7 TVD-12 275 320 325 340 355 335 360
8 TVD-17 282 300 315 280 245 260 340
9 TVD-20 190 220 195 225 240 265 300
The effect of various organic and inorganic nitrogen sources
on protease production was also studied and the results are
presented in Table 3. Among the various nitrogen sources,
high yield of protease production was observed in yeast
extract ranging between 360-475 U/ml in TVD-4, TVD-5,
TVD-6, TVD-9, TVD-12, and TVD-17 strains. However,
TVD-8 and TVD-20 showed maximum protease production
(425 and 280 U/ml) in presence of beef extract followed by
TVD-11 which used urea as a nitrogen source to achieve
maximum protease production of 320 U/ml. Many
researchers have reported that organic nitrogen sources were
better suited to Bacillus sp. for growth and enzyme
production than inorganic sources [1, 3]. The best nitrogen
source for protease production was beef extract for Bacillus
sp., while yeast extract and tryptone were comparable [1].
Several researchers have also reported that organic nitrogen
sources were found better for enzyme production than
inorganic sources [3, 14].
Table-3: Effect of various nitrogen sources on protease production of selected bacteria (Unit/ml)
S.No.
Bacterial
strains
Maximum Protease production (Unit/ml) after 24 h incubation
Beef
extract
Yeast
extract
Urea
Ammonium
nitrate
Sodium
nitrate
Ammonium
sulphate
1 TVD-4 330 350 310 300 295 320
2 TVD-5 390 425 405 370 355 400
3 TVD-6 365 375 335 320 315 340
4 TVD-8 425 408 410 310 343 350
5 TVD-9 445 475 365 390 380 415
6 TVD-11 245 270 310 220 243 260
7 TVD-12 355 380 326 275 290 310
8 TVD-17 310 315 330 440 280 460
9 TVD-20 280 245 232 205 230 240
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4. CONCLUSION
In this paper, we reported the isolation and screening of
bacteria from tannery solid waste which is chromate
resistant and capable to produce haloalkaliphilic protease.
Nine strains were found to be tolerant to high concentration
of Cr(VI) (850-1450 mg/l) and NaCl (6.5-9%) and was
capable of producing proteolytic enzyme under highly
alkaline conditions. Interestingly, these bacteria were able to
produce haloalkaline protease under wide temperature (25-
45°C) and pH (8.0-10.5) range. Further, significant protease
production was obtained within 24 h in presence of wheat
bran as a carbon source and yeast extract or beef extract as
nitrogen source. The potential of these strains to grow on
cheap carbon source and also producing high amount of
haloalkaline protease in short time makes it a promising
candidate for industrial application and environmental
restoration such as treatment of chromium contaminated
proteinaceous saline wastes.
ACKNOWLEDGEMENTS
Authors are extremely thankful to the “Science and
Engineering Research Board” (SERB), Department of
Science and Technology (DST), Government of India
(GOI), New Delhi for financial support as “Young Scientist
Research Project” for this work.
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BIOGRAPHIES
Dr. Tuhina Verma, Assistant Professor,
Department of Microbiology, Dr. Ram
Manohar Lohia, Avadh University,
Faizabad,- 224 001 Uttar Pradesh, India,
tuhinaverma19@gmail.com
Ms. Swati Agarwal, Research Scholar,
Department of Microbiology, Dr. Ram
Manohar Lohia, Avadh University,
Faizabad,- 224 001 Uttar Pradesh, India,
swati1224@gmail.com