Bioprocess development for enhanced spore production in shake flask and pilot...iosrjce
Bacillus thuringiensis subsp. Israelensis (Bti), has proven to be a safe and effective larvicide for controlling
mosquito and black fly larvae. The effect of cultivation and bioprocess development on Bti growth and sporulation was
investigated in shake flask level and batch cultivation in the semi-industrial scale 16-L stirred tank bioreactor. For
industrial production of biocontrol microorganism, it is necessary to obtain high cell mass and spore production in a short
time with low cost cultivation media. In this study, the new composition of production media was optimized which composed
of (g L
-1
): glucose, 10; yeast extract, 30; KH2PO4, 5; K2HPO4, 5; MgSO4. 7H20, 0.005; MnSO4.H2O, 0.03; FeSO4, 0.01;
CaCl2.7 H2O, 0.05; NaH2PO4, 1.5; NH4H2PO4, 1.5. The maximal cell dry mass and spore production, Sporemax for shake
flask study were 4.26 gL-1
at 36 h and 3.29106
spore mL-1
, respectively. Furthermore, studies of the cultivation conditions
under controlled and uncontrolled pH in the 16L-bioreactor was performed. The growth of Bti under uncontrolled pH
cultivation showing decreased of glucose and total protein concentration in the media was correlated with the vegetative cell
growth and sporulation. The maximal cell dry mass and Sporemax for uncontrolled pH bioreactor were 4.14 gL-1
at 36h and
3.7106
spore mL-1
, respectively. The maximal cell dry mass and spore production, Sporemax for controlled pH bioreactor
were 3.36 gL-1
at 26 h and 3.23 106
spore mL-1
, respectively. In conclusion, batch cultivation in 16-L bioreactor with the
new optimized production medium under uncontrolled pH condition increased of the cell dry mass and number of spores up to 23 % and 47 % , respectively
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.
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.
Screening, identification and isolation of cellulolytic fungiDr. sreeremya S
Cellulase assay for Enzyme production
The activity of -glucosidase ( G), total
cellulase (FPase) and endoglucanase
(CMCase) was studied as cellulolytic
activity. Filter paper activity (FPase) for
total cellulase activity in the culture filtrate
was determined according to the standard
method (Eveleigh DE et al.2009). CMCase
(carboxy methyl cellulase) activity was
assayed using Dinitrosalicylic acid (DNS)
method (Mandels and Weber, 1969).
Phytochemical and acute toxicity study of leaves of artocarpus heterophyllus lampharmaindexing
Artocarpus heterophyllus Lam commonly known as jack fruit widely distributed in north east India, West Bengal and south Karnataka. In the present study intended with various phytochemical screening and toxicity studies were carried out on the leaves of Artocarpus Heterophyllus. The phytochemical study shows the presences of flavonoids, tannins, saponins and carbohydrates in methanolic and aqueous extracts. In acute toxicity study both the extract were found safe on a dose of 2000 mg/Kg.
Spore Forming Bacterium from Oil Contaminated Soil as a Source of a Lipase En...IOSRJPBS
Twenty two bacterial isolates were obtained from oil contaminated soil, collected from some oil stations in Jeddah. All the obtained bacterial isolates were screened on Tween-Yeast extract medium for lipase production. Three bacterial isolates HM10, HM15 and HM20 showed the highest growth and lipase production agar medium, thus they were grown in liquid olive oil medium at 120 rpm. Maximum lipase production was obtained by the isolate HM10. The isolate HM10 was characterized and identified through physiological, biochemical tests and culture characteristics in addition to 16S rDNA as Bacillus coagulans. The effects of different factors on the enzyme production were studied. It was found that bacterial growth in medium 4 at initial pH 7.0, containing olive oil and incubation at 37ºC for 2 days at 120 rpm were the most favorable conditions for maximum lipase production by the tested isolate. The bacterial isolate was grown using the best culture conditions and lipase was precipitated using 80% of ammonium sulphate, purified using colum chromatography and characterized. The molecular weight was 62 kDa and the maximum enzyme activity was at 50ºC and pH 7.0. Presence of K + and Ca++ ions enhance enzyme activity.
Bioprocess development for enhanced spore production in shake flask and pilot...iosrjce
Bacillus thuringiensis subsp. Israelensis (Bti), has proven to be a safe and effective larvicide for controlling
mosquito and black fly larvae. The effect of cultivation and bioprocess development on Bti growth and sporulation was
investigated in shake flask level and batch cultivation in the semi-industrial scale 16-L stirred tank bioreactor. For
industrial production of biocontrol microorganism, it is necessary to obtain high cell mass and spore production in a short
time with low cost cultivation media. In this study, the new composition of production media was optimized which composed
of (g L
-1
): glucose, 10; yeast extract, 30; KH2PO4, 5; K2HPO4, 5; MgSO4. 7H20, 0.005; MnSO4.H2O, 0.03; FeSO4, 0.01;
CaCl2.7 H2O, 0.05; NaH2PO4, 1.5; NH4H2PO4, 1.5. The maximal cell dry mass and spore production, Sporemax for shake
flask study were 4.26 gL-1
at 36 h and 3.29106
spore mL-1
, respectively. Furthermore, studies of the cultivation conditions
under controlled and uncontrolled pH in the 16L-bioreactor was performed. The growth of Bti under uncontrolled pH
cultivation showing decreased of glucose and total protein concentration in the media was correlated with the vegetative cell
growth and sporulation. The maximal cell dry mass and Sporemax for uncontrolled pH bioreactor were 4.14 gL-1
at 36h and
3.7106
spore mL-1
, respectively. The maximal cell dry mass and spore production, Sporemax for controlled pH bioreactor
were 3.36 gL-1
at 26 h and 3.23 106
spore mL-1
, respectively. In conclusion, batch cultivation in 16-L bioreactor with the
new optimized production medium under uncontrolled pH condition increased of the cell dry mass and number of spores up to 23 % and 47 % , respectively
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.
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.
Screening, identification and isolation of cellulolytic fungiDr. sreeremya S
Cellulase assay for Enzyme production
The activity of -glucosidase ( G), total
cellulase (FPase) and endoglucanase
(CMCase) was studied as cellulolytic
activity. Filter paper activity (FPase) for
total cellulase activity in the culture filtrate
was determined according to the standard
method (Eveleigh DE et al.2009). CMCase
(carboxy methyl cellulase) activity was
assayed using Dinitrosalicylic acid (DNS)
method (Mandels and Weber, 1969).
Phytochemical and acute toxicity study of leaves of artocarpus heterophyllus lampharmaindexing
Artocarpus heterophyllus Lam commonly known as jack fruit widely distributed in north east India, West Bengal and south Karnataka. In the present study intended with various phytochemical screening and toxicity studies were carried out on the leaves of Artocarpus Heterophyllus. The phytochemical study shows the presences of flavonoids, tannins, saponins and carbohydrates in methanolic and aqueous extracts. In acute toxicity study both the extract were found safe on a dose of 2000 mg/Kg.
Spore Forming Bacterium from Oil Contaminated Soil as a Source of a Lipase En...IOSRJPBS
Twenty two bacterial isolates were obtained from oil contaminated soil, collected from some oil stations in Jeddah. All the obtained bacterial isolates were screened on Tween-Yeast extract medium for lipase production. Three bacterial isolates HM10, HM15 and HM20 showed the highest growth and lipase production agar medium, thus they were grown in liquid olive oil medium at 120 rpm. Maximum lipase production was obtained by the isolate HM10. The isolate HM10 was characterized and identified through physiological, biochemical tests and culture characteristics in addition to 16S rDNA as Bacillus coagulans. The effects of different factors on the enzyme production were studied. It was found that bacterial growth in medium 4 at initial pH 7.0, containing olive oil and incubation at 37ºC for 2 days at 120 rpm were the most favorable conditions for maximum lipase production by the tested isolate. The bacterial isolate was grown using the best culture conditions and lipase was precipitated using 80% of ammonium sulphate, purified using colum chromatography and characterized. The molecular weight was 62 kDa and the maximum enzyme activity was at 50ºC and pH 7.0. Presence of K + and Ca++ ions enhance enzyme activity.
Isolation Characterization and Screening of fungal Lipase from oil contaminat...AI Publications
Present scenario demands a more sustainable, ecofriendly and economic measures globally to deal with the growing problems of environmental issues. The main goal of this work is to opt for such ideas and technologies which involve cleaner and greener procedures for utilizing waste materials for deriving value added products. The soil pertaining to the areas of oil mills contains densely population of various microbes’, especially fungal origin. These microbes are rich in lipase content (due to oil source). Thus in this we isolated fungal colonies from this oil rich soil, cultured in laboratory, fermented them under various conditions to extract fungal enzyme i.e. lipase and then used it for further applications. Lipases are highly versatile and industrially important enzymes. Deriving the lipases from waste soil is the main attraction of this work and is a venture strategizing the “best from waste” approach.
Potential role of microbial surfactants in environment control recovered from...SUS GROUP OF INSTITUTIONS
A total of 20 samples were collected from contaminated (oil contaminated) as well as non-contaminated (agricultural) sites. A
total of 10 bacterial isolates were recovered from these samples out of which 6 were recovered from non contaminated sites
and 4 were recovered from contaminated sites gave emulsification index ranged from 44% to 73%. Different carbon sources
viz. maltose, starch, sucrose, mannitol and nitrogen sources viz. urea, peptone, potassium nitrate and ammonium nitrate
were screened to obtain optimum emulsification activity by KMSS09 and KIWS11. In this study mannitol and peptone was
evaluated as best carbon and nitrogen source for the production of bioemulsifier. Further these potential isolates were
evaluated for some environmental applications viz. Microbial Enhanced Oil Recovery and Bacterial Adhesion to Hydrocarbon
assay having important role in bioremediation. The percentage oil recovered by KMSS09, KIWS11 and P. aeruginosa MTCC
2297 was 51.67%, 71.67% and 85.0% respectively. In BATH assay, percentage of bacterial adherence by KMSS09, KIWS11
and P. aeruginosa MTCC 2297 was 80.4%, 86.3% and 93.2% respectively showing wide applicability in bioremediation for
pollution remediation of metal and hydrocarbon contaminated field.
IJERA (International journal of Engineering Research and Applications) is International online, ... peer reviewed journal. For more detail or submit your article, please visit www.ijera.com
Anti-Microbial Activity of probiotic Lactobacilli and Optimization of Bacteri...ijsrd.com
The present study is about the anti-microbial activity of the bacteriocin producing lactobacilli and optimization of bacteriocin production. Bacteriocin was extracted by solvent extraction with chloroform and the antimicrobial activity was tested against 5 different pathogens by agar spotting method. Optimization of bacteriocin production was done for 4 different parameters such as pH, Temperature, Carbon source and Nitrogen source and the anti-microbial activity was tested against the following 5 different pathogens and the results were observed and diameter of the zone of inhibition was measured and tabulated. From the results of the study it was found that bacteriocin produced from lactobacilli has good antimicrobial activity.The present study is about the anti-microbial activity of the bacteriocin producing lactobacilli and optimization of bacteriocin production. Bacteriocin was extracted by solvent extraction with chloroform and the antimicrobial activity was tested against 5 different pathogens by agar spotting method. Optimization of bacteriocin production was done for 4 different parameters such as pH, Temperature, Carbon source and Nitrogen source and the anti-microbial activity was tested against the following 5 different pathogens and the results were observed and diameter of the zone of inhibition was measured and tabulated. From the results of the study it was found that bacteriocin produced from lactobacilli has good antimicrobial activity.
Isolation and Molecular Characterization of Pullulanase Producing Bacillus St...YogeshIJTSRD
Pullulanase is an extracellular carbohydrase responsible for the hydrolysis of pullulan and amylopectin toproduce maltotriose. The product maltotriose is used in detergent industry, bakery industry and in the production of biotechnological products. In the present investigation pullulanase producing bacillus species were isolated and characterized using different biochemical and molecular methodologies. The isolates were identified as Bacillus cereus and Bacillus thuringiensis respectively.. The pullulanase acivity was higher in Bacillus cereus, 0.62U ml than B. thuringiensis, 0.53 U ml. This research reveals that pullulanase enzyme production from these Bacillus species shows great promise for use in industrial processes. Nwozor, N. C | Ogbo, F. C "Isolation and Molecular Characterization of Pullulanase Producing Bacillus Strains" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd45051.pdf Paper URL: https://www.ijtsrd.com/biological-science/microbiology/45051/isolation-and-molecular-characterization-of-pullulanase-producing-bacillus-strains/nwozor-n-c
Isolation and characterization of lipolytic Pseudomonas spp. from oil contami...iosrjce
Oil contaminated water samples collected from different areas in Pune were screened for the
selective isolation of lipolytic Pseudomonas spp. Screening medium containing Tributyrin (1% v/v) and
Rhodamine B was used for the specific isolation of lipolytic Pseudomonas spp. from oil contaminated water
samples. Lipolytic bacteria showing zone of clearance around the colonies were selected for the present study.
Isolated bacteria were identified upto genus level with the help of morphological and biochemical testing by
following Bergey’s manual. Spectrophotometric Lipase assay showed that out of 49 lipolytic Pseudomonas spp.,
WP23 exhibits the maximum Lipase activity of about 45.4733 U/ml.
Effect of Different Physico-Chemical Parameters on Production ofAmylase by Ba...IOSR Journals
The present study is concerned with the production of amylase by Bacillus species strain. In this
study 12 bacterial strains were isolated and screened for their α-amylase activity. These strains were
maintained on nutrient agar medium. Fermentation for the production of amylase was carried out in the enzyme
production medium (EPM). All the 12 strains were tested for amylase production. On the basis of maximum
amylase activity strain no.1 was selected for further studies. Different starch concentrations, 0.75,1.00,1.25%,
pH labels 6.5,7.0,7.5,8.0, aeration (RPM), 100,120,140, temperatures 250C,280C,370C, and 400C and inoculums
level 0.5%,1.0%, 1.5% and 2.0% were studied
Secondary screening of industrial important microbes DhruviSuvagiya
Detection and isolation of a microorganism from a natural environment like soil containing large number of microbial population is called as screening. It is very time consuming and expensive process.
Optimum Conditions for Alginaseby Bacilllus Circulans R Isolateiosrphr_editor
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
CULTURE METHODS
Indications for culture -
Isolate bacteria in pure cultures.
Demonstrate their properties.
Obtain sufficient growth for preparation of antigens & for other tests.
Typing bacterial isolates.
Antibiotic sensitivity.
Estimate viable counts.
Maintain stock cultures.
Streak culture or surface plating
Lawn or carpet culture
Stroke culture
Stab culture
Pour plate method
Anaerobic methods of culturing bacteria
Streak Culture
Routinely employed for isolation
Platinum / Nichrome loops
LAWN OR CARPET CULTURE
Uniform surface growth
Bacteriophage typing
Antibiotic sensitivity testing
Preparation of bacterial antigens & vaccines
STOKE CULTURE
Tubes containing agar slopes
For slide agglutination & other diagnostic tests.
STAB CULTURE
By puncturing a suitable medium with a long, straight charged wire.
For gelatin liquefaction, stock cultures & motility
POUR PLATE METHOD
1 ml of appropriately diluted inoculum is added to 15 ml of molten agar and poured on petridish.
Colonies appear through out the depth of medium.
Used to estimate viable count, recommended method for quantitative urine cultures.
BROTH CULTURE
Inoculated by a charged loop, pipette or syringes.
For blood cultures & sterility testing.
Biodegradation Potentials of Aspergillus Flavipes Isolated from Uburu and Okp...YogeshIJTSRD
Saline lakes are water bodies with salinity greater than 3 g l 0.3 , while hypersaline lakes are water bodies that surpass the moderate 35 g l 3.5 salt of oceans. Hypersaline lakes, could either be thalassohaline which are creations of evaporation of seawater and as such contain sodium chloride as the major salt, with a salinity that surpasses that of seawater by a factor of 5 – 10, with a neutral or slightly alkaline pH . Whereas, athalassohaline lakes stem from non seawater sources and are made up of high concentrations of ions such as magnesium and calcium and sundry other ions such as potassium, or sodium in smaller amounts. This work has revealed the biodegradation potentials of some halophiles isolated from Uburu and Okposi salt lakes. The isolates recovered in descending order of salt tolerance were Aspergillus flavipes 13mm at 40 , Penicillium citrinum 10mm at 40 , Aspergillus ochraceus 9mm at 40 , Aspergillus nomius 15mm at 35 , Microsphaeropsis arundinis 12mm at 35 , Aspergillus sydowi 28mm at 30 , Penicillium janthinellum 26mm at 30 , Mucor sp 13mm at 30 , Aureobasidium sp 12mm at 30 , Trichoderma sp 9mm at 30 , Alternaria sp. 22mm at 25 , Aspergillus sp 18mm at 25 , Penicillium sp 20mm at 20 , Cladosporium sp. 7mm at 15 and identified using ITS rDNA Sequencing Macrogen, South Korea . They belonged to the borderline extreme halophiles and moderate halophiles respectively. The biodegradative potential of Aspergillus flavipes was ascertained by testing it against 2 , 4 and 6 crude oil and it grew only on 2 crude oil Bushnell Haas broth with a fungal count of 2.56x105 cfu ml. Crude oil degradation rate was evaluated biweekly gravimetrically with 22 degradation in 2 weeks, 36 in 4 weeks, 67 in 6 weeks and 89 in 8 weeks as well as by way of gas chromatography GC FID , which showed that fractions C10 C11 were significantly degraded, C12 C20, moderately degraded and C26 C34, insignificantly degraded. Kingsley C. Agu | Frederick J. C. Odibo "Biodegradation Potentials of Aspergillus Flavipes Isolated from Uburu and Okposi Salt Lakes" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd44949.pdf Paper URL: https://www.ijtsrd.com/biological-science/microbiology/44949/biodegradation-potentials-of-aspergillus-flavipes-isolated-from-uburu-and-okposi-salt-lakes/kingsley-c-agu
Isolation Characterization and Screening of fungal Lipase from oil contaminat...AI Publications
Present scenario demands a more sustainable, ecofriendly and economic measures globally to deal with the growing problems of environmental issues. The main goal of this work is to opt for such ideas and technologies which involve cleaner and greener procedures for utilizing waste materials for deriving value added products. The soil pertaining to the areas of oil mills contains densely population of various microbes’, especially fungal origin. These microbes are rich in lipase content (due to oil source). Thus in this we isolated fungal colonies from this oil rich soil, cultured in laboratory, fermented them under various conditions to extract fungal enzyme i.e. lipase and then used it for further applications. Lipases are highly versatile and industrially important enzymes. Deriving the lipases from waste soil is the main attraction of this work and is a venture strategizing the “best from waste” approach.
Potential role of microbial surfactants in environment control recovered from...SUS GROUP OF INSTITUTIONS
A total of 20 samples were collected from contaminated (oil contaminated) as well as non-contaminated (agricultural) sites. A
total of 10 bacterial isolates were recovered from these samples out of which 6 were recovered from non contaminated sites
and 4 were recovered from contaminated sites gave emulsification index ranged from 44% to 73%. Different carbon sources
viz. maltose, starch, sucrose, mannitol and nitrogen sources viz. urea, peptone, potassium nitrate and ammonium nitrate
were screened to obtain optimum emulsification activity by KMSS09 and KIWS11. In this study mannitol and peptone was
evaluated as best carbon and nitrogen source for the production of bioemulsifier. Further these potential isolates were
evaluated for some environmental applications viz. Microbial Enhanced Oil Recovery and Bacterial Adhesion to Hydrocarbon
assay having important role in bioremediation. The percentage oil recovered by KMSS09, KIWS11 and P. aeruginosa MTCC
2297 was 51.67%, 71.67% and 85.0% respectively. In BATH assay, percentage of bacterial adherence by KMSS09, KIWS11
and P. aeruginosa MTCC 2297 was 80.4%, 86.3% and 93.2% respectively showing wide applicability in bioremediation for
pollution remediation of metal and hydrocarbon contaminated field.
IJERA (International journal of Engineering Research and Applications) is International online, ... peer reviewed journal. For more detail or submit your article, please visit www.ijera.com
Anti-Microbial Activity of probiotic Lactobacilli and Optimization of Bacteri...ijsrd.com
The present study is about the anti-microbial activity of the bacteriocin producing lactobacilli and optimization of bacteriocin production. Bacteriocin was extracted by solvent extraction with chloroform and the antimicrobial activity was tested against 5 different pathogens by agar spotting method. Optimization of bacteriocin production was done for 4 different parameters such as pH, Temperature, Carbon source and Nitrogen source and the anti-microbial activity was tested against the following 5 different pathogens and the results were observed and diameter of the zone of inhibition was measured and tabulated. From the results of the study it was found that bacteriocin produced from lactobacilli has good antimicrobial activity.The present study is about the anti-microbial activity of the bacteriocin producing lactobacilli and optimization of bacteriocin production. Bacteriocin was extracted by solvent extraction with chloroform and the antimicrobial activity was tested against 5 different pathogens by agar spotting method. Optimization of bacteriocin production was done for 4 different parameters such as pH, Temperature, Carbon source and Nitrogen source and the anti-microbial activity was tested against the following 5 different pathogens and the results were observed and diameter of the zone of inhibition was measured and tabulated. From the results of the study it was found that bacteriocin produced from lactobacilli has good antimicrobial activity.
Isolation and Molecular Characterization of Pullulanase Producing Bacillus St...YogeshIJTSRD
Pullulanase is an extracellular carbohydrase responsible for the hydrolysis of pullulan and amylopectin toproduce maltotriose. The product maltotriose is used in detergent industry, bakery industry and in the production of biotechnological products. In the present investigation pullulanase producing bacillus species were isolated and characterized using different biochemical and molecular methodologies. The isolates were identified as Bacillus cereus and Bacillus thuringiensis respectively.. The pullulanase acivity was higher in Bacillus cereus, 0.62U ml than B. thuringiensis, 0.53 U ml. This research reveals that pullulanase enzyme production from these Bacillus species shows great promise for use in industrial processes. Nwozor, N. C | Ogbo, F. C "Isolation and Molecular Characterization of Pullulanase Producing Bacillus Strains" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd45051.pdf Paper URL: https://www.ijtsrd.com/biological-science/microbiology/45051/isolation-and-molecular-characterization-of-pullulanase-producing-bacillus-strains/nwozor-n-c
Isolation and characterization of lipolytic Pseudomonas spp. from oil contami...iosrjce
Oil contaminated water samples collected from different areas in Pune were screened for the
selective isolation of lipolytic Pseudomonas spp. Screening medium containing Tributyrin (1% v/v) and
Rhodamine B was used for the specific isolation of lipolytic Pseudomonas spp. from oil contaminated water
samples. Lipolytic bacteria showing zone of clearance around the colonies were selected for the present study.
Isolated bacteria were identified upto genus level with the help of morphological and biochemical testing by
following Bergey’s manual. Spectrophotometric Lipase assay showed that out of 49 lipolytic Pseudomonas spp.,
WP23 exhibits the maximum Lipase activity of about 45.4733 U/ml.
Effect of Different Physico-Chemical Parameters on Production ofAmylase by Ba...IOSR Journals
The present study is concerned with the production of amylase by Bacillus species strain. In this
study 12 bacterial strains were isolated and screened for their α-amylase activity. These strains were
maintained on nutrient agar medium. Fermentation for the production of amylase was carried out in the enzyme
production medium (EPM). All the 12 strains were tested for amylase production. On the basis of maximum
amylase activity strain no.1 was selected for further studies. Different starch concentrations, 0.75,1.00,1.25%,
pH labels 6.5,7.0,7.5,8.0, aeration (RPM), 100,120,140, temperatures 250C,280C,370C, and 400C and inoculums
level 0.5%,1.0%, 1.5% and 2.0% were studied
Secondary screening of industrial important microbes DhruviSuvagiya
Detection and isolation of a microorganism from a natural environment like soil containing large number of microbial population is called as screening. It is very time consuming and expensive process.
Optimum Conditions for Alginaseby Bacilllus Circulans R Isolateiosrphr_editor
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
CULTURE METHODS
Indications for culture -
Isolate bacteria in pure cultures.
Demonstrate their properties.
Obtain sufficient growth for preparation of antigens & for other tests.
Typing bacterial isolates.
Antibiotic sensitivity.
Estimate viable counts.
Maintain stock cultures.
Streak culture or surface plating
Lawn or carpet culture
Stroke culture
Stab culture
Pour plate method
Anaerobic methods of culturing bacteria
Streak Culture
Routinely employed for isolation
Platinum / Nichrome loops
LAWN OR CARPET CULTURE
Uniform surface growth
Bacteriophage typing
Antibiotic sensitivity testing
Preparation of bacterial antigens & vaccines
STOKE CULTURE
Tubes containing agar slopes
For slide agglutination & other diagnostic tests.
STAB CULTURE
By puncturing a suitable medium with a long, straight charged wire.
For gelatin liquefaction, stock cultures & motility
POUR PLATE METHOD
1 ml of appropriately diluted inoculum is added to 15 ml of molten agar and poured on petridish.
Colonies appear through out the depth of medium.
Used to estimate viable count, recommended method for quantitative urine cultures.
BROTH CULTURE
Inoculated by a charged loop, pipette or syringes.
For blood cultures & sterility testing.
Biodegradation Potentials of Aspergillus Flavipes Isolated from Uburu and Okp...YogeshIJTSRD
Saline lakes are water bodies with salinity greater than 3 g l 0.3 , while hypersaline lakes are water bodies that surpass the moderate 35 g l 3.5 salt of oceans. Hypersaline lakes, could either be thalassohaline which are creations of evaporation of seawater and as such contain sodium chloride as the major salt, with a salinity that surpasses that of seawater by a factor of 5 – 10, with a neutral or slightly alkaline pH . Whereas, athalassohaline lakes stem from non seawater sources and are made up of high concentrations of ions such as magnesium and calcium and sundry other ions such as potassium, or sodium in smaller amounts. This work has revealed the biodegradation potentials of some halophiles isolated from Uburu and Okposi salt lakes. The isolates recovered in descending order of salt tolerance were Aspergillus flavipes 13mm at 40 , Penicillium citrinum 10mm at 40 , Aspergillus ochraceus 9mm at 40 , Aspergillus nomius 15mm at 35 , Microsphaeropsis arundinis 12mm at 35 , Aspergillus sydowi 28mm at 30 , Penicillium janthinellum 26mm at 30 , Mucor sp 13mm at 30 , Aureobasidium sp 12mm at 30 , Trichoderma sp 9mm at 30 , Alternaria sp. 22mm at 25 , Aspergillus sp 18mm at 25 , Penicillium sp 20mm at 20 , Cladosporium sp. 7mm at 15 and identified using ITS rDNA Sequencing Macrogen, South Korea . They belonged to the borderline extreme halophiles and moderate halophiles respectively. The biodegradative potential of Aspergillus flavipes was ascertained by testing it against 2 , 4 and 6 crude oil and it grew only on 2 crude oil Bushnell Haas broth with a fungal count of 2.56x105 cfu ml. Crude oil degradation rate was evaluated biweekly gravimetrically with 22 degradation in 2 weeks, 36 in 4 weeks, 67 in 6 weeks and 89 in 8 weeks as well as by way of gas chromatography GC FID , which showed that fractions C10 C11 were significantly degraded, C12 C20, moderately degraded and C26 C34, insignificantly degraded. Kingsley C. Agu | Frederick J. C. Odibo "Biodegradation Potentials of Aspergillus Flavipes Isolated from Uburu and Okposi Salt Lakes" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd44949.pdf Paper URL: https://www.ijtsrd.com/biological-science/microbiology/44949/biodegradation-potentials-of-aspergillus-flavipes-isolated-from-uburu-and-okposi-salt-lakes/kingsley-c-agu
Screening of Biosurfactant Bioemulsifier Producing Bacteria from Petroleum Co...ijtsrd
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Screening and characterization of biosurfactants producing microorganism form natural environment(whey spilled soil)
1. Journal of Natural Sciences Research
ISSN 2224-3186 (Paper) ISSN 2225-0921 (Online)
Vol.3, No.13, 2013
www.iiste.org
Screening and Characterization of Biosurfactants Producing
Microorganism formNatural Environment(Whey Spilled Soil)
1.
2.
Shaikh Rajesh Ali1*Banani Ray Chowdhury2Prajna Mondal1Sisir Rajak1
Department of Microbiology, AcharyaPrafulla Chandra College, New Barrackpore, Kolkata-700131,
West Bengal, India
Department of Biotechnology, West Bengal University of Technology, BF 142 Sector I Salt Lake
Kolkata 700 064, West Bengal, India
* E-mail of the corresponding author: raaz_boseinst@yahoo.co.in
Abstract:
Biosurfactant are surface active agent produce by diverse group of microorganism. Disposal of whey is common
in India especially in West Bengal, so in our study we collect sample form different whey spilled soil and isolate
biosurfactant producing bacteria. Total ten different types of bacteria (APCCS1a-S5b) we have isolate for our
study. Among them four are screened by growing them in CTAB agar plates and their hemolytic activity in
blood agar plates. We analyses four strains for their ability to produce extracellular biosurfactant by oil spreading
technique and finally confirmed by drop collapsing test. The extracellular biosurfactant of the isolated strains
were extracted by conventional way and they are characterized by thin layer chromatography (TLC) and were
shown that they are glycolipid (APCCS1a, APCCS3b and APCCS5a) and lipopeptide (APCCS1b) in nature.The
surface tension activities of extracted biosurfactant were evaluated by emulsification index (E24). Finally we
characterized the isolated strains according to Berge`s Manual and they are belonging to Pseudomonas
(APCCS1a and APCCS5a), Lactobacillus (APCCS2b) and Bacillus (APCCS1b) group.
Keywords: Biosurfactants, Thin layer chromatography (TLC), cetyltrimethylammonium bromide (CTAB),
hemolysis, whey, emulsification index.
1. Introduction
Biosurfactants are surface active molecules having hydrophilic and hydrophobic moieties as their constituents
which allow them to interact at interfaces and reduce the surface tension. They are produce by diverse group of
organism belong to bacteria, fungi and actinomycetesetc., mainly on surfaces of microorganisms or may also
secreted extracellularly. They are categorized based on their chemical composition as fatty acids, glycolipids,
glycolipopeptides, glycoproteins, lipopeptides, phospholipids, polymeric and particulate biosurfactants. The
chemicaldiversity of biosurfactants makes them a potential source for green chemicals having applications in
industrial, environmental (agricultural and bioremediation), and medical fields. Almost all surfactants being
currently produced are derived from petroleum source. However, these synthetic surfactants are usually toxic
and hardly degraded by microorganisms. These are potential source of pollution and damage to the environment.
Therefore, in the recent years, much interest and attention have been directed towards biosurfactants over
chemically synthesized surfactants due to their superiority to the chemical surfactants with respect to their
biocompatibility, lower toxicity, higher biodegradability, higher stability, extreme stability in extreme
temperature and pH. With the advent of time, this attribute is contributing its higher demand in the field of
biotechnology.
The agricultural waste such as whey (a by-product of the manufacture of cheese or casein) are well
known for containing high levels of carbohydrates and of lipids -both of which are necessary for substrates for
the production of biosurfactants and contains all necessary substances (lactose, protein, organic acids and
vitamins) that require for growth of surfactant producing microorganism. This study focus on the screening,
production, extraction and purification of biosurfactant from bacteria isolated from whey spilled soil and which
is easily available in India.
2. Materials and methods
2.1. Sampling area
For isolation biosurfactant producing bacteria soil samples were collected from whey spilled surfaces of five
different cheese making farm of West Bengal, India (sample 1-5). The samples were collected in sterile
container under aseptic condition and were taken to the laboratory for analysis. The pH of the samples during
collection was 7.0 and temperature was 300C.
2.2. Enrichment, Isolation and enumeration of bacterial isolates
5.0g of the whey spilled soil samples were dissolving in 100 ml of phosphate buffer saline (PBS). After
precipitation of solid debris 5ml liquid suspension are inoculated in 50ml of nutrient broth and incubated at 25°C
with agitation speed of 200 rpm for 48 hours.After incubation the medium was serially diluted from 10-1 to 10-6
53
2. Journal of Natural Sciences Research
ISSN 2224-3186 (Paper) ISSN 2225-0921 (Online)
Vol.3, No.13, 2013
www.iiste.org
in sterile water. From the dilutions (10-1 to 10-6) 1ml was transferred to sterile petri-dish containing 20mls of
Reasoner´s 2A agar (R2A) contained (g/L): Proteose peptone, 0.5; Casamino acids, 0.5; Yeast extract, 0.5;
Dextrose, 0.5; Soluble starch, 0.5; Dipotassium phosphate, 0.3; Magnesium sulfate 7H2O, 0.05; Sodium pyruvate,
0.3; Agar, 15; Final pH 7.2 ± 0.2 @ 25 °C, by spread plate techniques. The plates were then inverted and
incubated at 25°C for 48 hours. Control and replica plates were maintained.
After incubation 30-300 colonies containing plates were selected and morphologically different
colonies were streaked on LB agar media and obtained pure culture by incubating at 370C for 24 hours. The pure
isolates were stored in R2A agar slants for further identification. These cultures were stored in R2A agar slants
and kept under refrigerated condition (40C) for further experimentation.
2.3. Screening of biosurfactant producing organisms
The isolated colonies were tested for their biosurfactant production by different methods; CTAB Agar Plate;Oil
Spreading Technique; Blood Hemolysis Test and Drop collapsing test.
2.3.1. CTAB Agar Plate
The CTAB agar plate method is a semi-quantitative assay for the detection of extra cellular glycolipids or other
anionic surfactants. It was developed by Siegmund and Wagner. Blue agar plates containing
cetyltrimethylammonium bromide (CTAB) (0.2 mg ml-1) and methylene blue (5 mg ml-1) were used to detect
extracellular glycolipid production. Biosurfactants were observed by the formation of dark blue halos around the
colonies.
2.3.2. Oil spreading technique
Isolate bacterial strains was incubated into 100 mL of culture medium. The Mckeen medium (20 gL-1 glucose,
5.0 gL-1 glutamic acid, 1.0 gL-1 K2HPO4, 1.02 gL-1 MgSO4, 0.5 gL-1KCl) supplemented with 1 mL of trace
elements solution (0.5 gL-1 MnSO4,.7H2O, 0.16 gL-1 CuSO4,.5H2O and 0.015 gL-1 FeSO4,.7H2O) adjusting
to pH 7.0 was used as cultural medium. The cultures were incubated on rotary shaker (150 rpm) for 3 days at
25 °C. The culture suspension was screened for biosurfactant production by the oil spreading techniques
(Anandaraj&Thivakaran, 2010; Priya&Usharani, 2009). The procedure is as follows: 30ml of distilled water
was taken in the petri dish (25cm in diameter). 20µl of crude oil was added to the center of the plates containing
distilled water. Now add 20µl of the supernatant of the culture suspension to the center. The biosurfactant
producing organism can displace the oil and spread in the water. The diameter and area of clear halo visualized
under visible light were measured and calculated after 1 minute.
2.3.3.Hemolytic activity test
Hemolytic activity appears to be a good screening criterion for surfactant-producing strains because
biosurfactant producing capacity was found to be associated with hemolytic activity. The fresh single colonies
from the isolated cultures were taken and streaked on blood agar plates. The plates were incubated for 48-72
hours at 37°C. Hemolytic activity was detected as the occurrence of a define clear zone around a colony (Carrillo
et al., 1996). These clear zones indicate the presence of bio-surfactants producing bacteria.
2.3.4.Drop collapsing test
Jain et al developed the drop collapse assay. This assay relies on the destabilization of liquid droplets by
surfactants. Therefore, drops of culture supernatant are placed on an oil coated, solid surface. If the liquid does
not contain surfactants, the polar water molecules are repelled from the hydrophobic surface and the drops
remain stable. If the liquid contains surfactants, the drops spread or even collapse because the force or interfacial
tension between the liquid drop and the hydrophobic surface is reduced. The stability of drops is dependent on
surfactant concentration and correlates with surface and interfacial tension. The assay was done in following way:
2µl of mineral oil was added to each well of a 96-well micro-titer plate lid. The lid was equilibrated for 1 hour at
room temperature, and then 5 µl of the cultural supernatant was added to the surface of oil. The shape of the drop
on the oil surface was inspected after 1 min. Biosurfactant-producing cultures giving flat drops were scored as
positive '+'. Those cultures that gave rounded drops were scored as negative '-', indicative of the lack of
biosurfactant production (Youssef et al., 2004).
2.4. Identification of bacteria
The isolated biosurfactant producing bacteria was characterizedby microbiological and biochemical tests such as
Gram staining, carbohydrate fermentation test, H2Sproduction test, indole production test, methylred test, VogesProskauer test, citrate utilization test, urease test,catalase test, oxidase test, litmus milk reaction, starch
hydrolysistest, gelatin hydrolysis test, lipid hydrolysis test.
Results from the biochemical analysis were used to find the closest match with known bacterial genus
and to assign the bacterial signature according to Bergey’s manual.
2.5. Extraction of biosurfactants
Each culture was inoculated in 50 ml of Mckeenbroth with 1 ml of whey. The culture was incubated at 250C for
3 days with shakingcondition. After incubation the bacterialcells were removed by centrifugation at 8000rpm,
40C for 10 minutes. Thesupernatant was taken and the pH of thesupernatant was adjusted to 2, using 1MH2SO4.
54
3. Journal of Natural Sciences Research
ISSN 2224-3186 (Paper) ISSN 2225-0921 (Online)
Vol.3, No.13, 2013
www.iiste.org
Now add equal volume ofchloroform: methanol (2:1). This mixturewas shaken well for mixing and left
overnight for evaporation. White colored sediment was obtained as a result i.e., the “Biosurfactants”.
2.6. Recharacterization of biosurfactants
Preliminary characterization of the biosurfactant was done by Thin Layer chromatography (TLC) method. A
spot of crude biosurfactant was placed on the silica plate(Merck & Co., Mumbai, India) and the biosurfactant
was separated on the plate using chloroform: methanol: water (:10:0.5). The plate was developed with different
color developing reagents. Ninhydrin reagent (0.5g ninhydrin in 100ml anhydrous acetone) was used to detect
lipopeptide biosurfactant as red spots and anthrone reagent (1g anthrone reagent in 5ml sulfuric acid mixed with
95 ml ethanol) to detect glycolipid biosurfactant as yellow spots (Yin et al., 2008).
2.7. Emulsification index (E24)
The emulsifying capacity was evaluated by an emulsification index (E24). The E24of culturesamples was
determined by adding 2 ml of kerosene and 2 ml of the cell-free broth in test tube, vortexing at high speed for 2
min and allowed to stand for 24h. The E index is given as percentage of the height of emulsified layer (cm)
divided by the total height of the liquid column (cm).The percentage of emulsification index calculated by using
the following equation (Cooper and Goldenberg, 1987).
3. Result
In the spread from the serial dilution tubes the colonies were enumerated and listed in the Table: 1. Ten different
types of colonies (2 from each samples) were identified morphologically and were purified and stored at 40C for
further analysis.
3.1. CTAB Agar Plate
CTAB Agar Plates results showed the dark blue halos in the six isolates (APCCS1a, 1b; APCCS2b; APCCS4a;
APCCS5a, 5b)indicating positive activity of Biosurfactant production (Table: 2).
3.2. Oil spreading technique
Cultures of ten isolated strains (APCCS1a, 1b; APCCS2a, 2b; APCCS3a, 3b; APCCS4a, 4b; APCCS5a, 5b) were
centrifuged and added to the oil containing plates. The strain APCCS1a, APCCS1b, APCCS3b and APCCS5a
showed the clear zone by being able to displace the oil around the colony indicating biosurfactant production. No
clear zone was observed with control. The results were tabulated (Table: 2).
3.3. Hemolytic Activity
The isolated were streaked in the blood agar plates. The hemolytic activity was observed in all the ten isolated
strains, results showed (alpha) hemolytic activity of strain APCCS2a, APCCS3b,APCCS4a,and APCCS5b, the
(beta) hemolytic activity of strain APCCS1a, APCCS1b, APCCS2band APCCS5aandthe (gamma) hemolytic
activity of strainAPCCS3aandAPCC4b(Table:2).
3.4. Drop collapsing test
The isolated culture supernatant are placed on an oil coated solid surface, the drops spread or even collapse
because the force or interfacial tension between the liquid drop and the hydrophobic surface is reduced indicate
the presence of surfactant in the cell supernatants (Table:2).
3.5. Characterization of biosurfactant producing organisms
The screened biosurfactant producing organism was taken for the extraction surfactants. First the organism was
identified by different biochemical tests. The results of it’s were tabulated (Table: 3). Comparing the results
with Bergey’sManual, identification of bacteria was performed.
3.6. Extraction of biosurfactants
The culture inoculated in Mckeen broth with whey was centrifuged and the supernatant was taken mixed with
Chloroform: methanol. White sediment was retained while after evaporation for overnight.
3.7. Characterization of biosurfactants
The crude biosurfactant produced were characterized by using silica thin layer chromatography (TLC) plates.
The sediment obtained was placed in the TLC plate and the plates when sprayed with ninhydrin reagent and
anthrone reagent it showed red spot (for APCCS1b) and yellow spots (forAPCCS1a,APCCS2b and APCCS5a) in
the plates respectively.
This shows the production of lipopeptide (for APCCS1b) and glycolipid
(forAPCCS1a,APCCS3b and APCCS5a)biosurfactants in the organisms.
3.8. Emulsification measurement
Emulsification activity was measured according to the method of Cooper and Goldenberg (1987). The
emulsification activity of isolated strain was measure after 24 hours indicate the value varies from 45% to 60%.
The results are given in table 4.
55
4. Journal of Natural Sciences Research
ISSN 2224-3186 (Paper) ISSN 2225-0921 (Online)
Vol.3, No.13, 2013
www.iiste.org
Table 1: ENUMERATION OF BACTERIAL STRAINS FROM THE SAMPLE
Dilution
Sample 1
Sample2
Sample 3
Sample 4
10-1
TNTC
TNTC
TNTC
TNTC
10-2
TNTC
TNTC
TNTC
TNTC
10-3
TNTC
TNTC
TNTC
TNTC
10-4
282
192
137
227
10-5
62
51
31
39
10-6
TFTC
TFTC
TFTC
TFTC
Average no of 4.51x106
3.51 x106
2.23 x106
3.08 x106
CFU/ml
Table 2: CHARACTERIZING BIOSURFACTANT PRODUCING BACTERIA
Isolated strains
Growth in CTAB
Haemolysis
Clear zone (mm)
agar Plates
On Oil spreading
technique
APCCS1a
++
β- hemolytic
12
APCCS1b
+
β- hemolytic
5
APCCS2a
α- hemolytic
No zone
APCCS2b
+
β- hemolytic
7
APCCS3a
γ - hemolytic
No zone
APCCS3b
α-hemolytic
No zone
APCCS4a
+
α- hemolytic
No zone
APCCS4b
γ - hemolytic
No zone
APCCS5a
++
β- hemolytic
11
APCCS5b
+
α-hemolytic
No zone
S.
mutans(+ve
+
α-hemolytic
No zone
control for αhemolytic)
P.aeruginosa(+ve
++
β- hemolytic
10
control for βhemolytic)
E.
coli
(+ve
γ- hemolytic
No zone
control for γhemolytic)
56
Sample5
TNTC
TNTC
TNTC
209
39
TFTC
2.99 x106
Drop collapsing test
+
+
+
+
-
+
-
5. Journal of Natural Sciences Research
ISSN 2224-3186 (Paper) ISSN 2225-0921 (Online)
Vol.3, No.13, 2013
www.iiste.org
Table 3: IDENTIFICATION OF THE BACTERIAL CULTURE
Character
APCCS1a
APCCS1b
Morphology
Rod shaped
Rod shaped
Gram staining
-ve
+ve
Endospore staining
-ve
+ve
Motility test
+
+
carbohydrate
Glucose
+
fermentation test
Maltose
+
+
lactose
sucrose
Fructose
Mannitol
H2S production test
indole production test
methyl red test
Voges-Proskauer test
+
citrate utilization test
+
+
urease test
+
catalase test
+
+
oxidase test
+
+
Nitrate reduction test
+
+
litmus milk reaction
+
+
starch hydrolysis test
+
gelatin hydrolysis test
+
lipid hydrolysis test
+
Strains Name (comparing with Pseudomonas sp. Bacillus sp.
Berge`s Manual)
APCCS2b
Rod shaped
+ve
-ve
+
+
+
+
+
+
+
+
Lactobacillus
sp.
APCCS5a
Rod shaped
-ve
-ve
+
+
+
+
+
+
+
+
+
+
+
+
Pseudomonas sp.
Table 4: EMULSIFICATION MEASUREMENT
Isolated strains
Emulsification index (E24)
APCCS1a
63±0.4
APCCS1b
33 ± 0.3
APCCS2b
41±0.3
APCCS5a
61±0.2
-veControl (E. coli)
00±0.2
+veControl
(P. 51±0.3
aeruginosa)
4. Discussion
Biosurfactant are potentially used recent years by different industry due to its nontoxic effects. The main aim of
our study to isolate and characterizedbiosurfactant producing bacteria form whey spilled soil. Whey spillage soil
contains all types of nutrient that require for growth of various types of microorganism along with bacteria those
responsible for surfactant production. We have been able to isolate bacteria with the ability to produce
biosurfactants from the soil samples collected from whey spillage area. The biosurfactants production ability of
isolated bacteria are confirmed by different screening method including CTAB agar plate method, oil spreading
technique, hemolytic activity and drop collapsing test.
Blue agar plate method is a semi quantitative agar plate method that is based on the formation of an
insoluble ion pair of anionic surfactants with the cationic surfactant CTAB and the basic dye methylene blue.
The isolated strains showed positive CTAB agar plate activity, indicate presence of extracellularbiosurfactant.
The oil spreading technique was used as indicator for biosurfactant production as reported by
Anandaraj&Thivakaran, 2010; Priya&Usharani, 2009 and successfully displace the oil was analyzed for isolated
strains. The strains are further confirming for their surfactant production ability by hemolytic activity and drop
collapsing test. All isolated four strain APCCS1a, APCCS1b, APCCS2b and APCCS5a give positive in hemolytic
activity among them first and second are beta hemolytic (complete) and third and fourth are alpha hemolytic
(incomplete). Finally drop collapsing test was done as by Youssef et al., 2004 of isolated culture and flat drops
were scored as positive '+' score biosurfactant production.
The strains are characterized using different morphological and biochemical process using Bergey’s
57
6. Journal of Natural Sciences Research
ISSN 2224-3186 (Paper) ISSN 2225-0921 (Online)
Vol.3, No.13, 2013
www.iiste.org
manual as reference. The isolated strains werebelonging to Pseudomonas (APCCS1a and APCCS5a), bacillus
(APCCS1b) and Lactobacillus (APCCS2b) groups. The surfactant from each of these four strains were extracted
by culturing in liquid medium (Mcknee medium) using convention technique and further characterized by thin
layer chromatography using Ninhydrin and anthrone reagent as developing solution. This shows the production
of lipopeptide (for APCCS1b) and glycolipid (forAPCCS1a, APCCS3b and APCCS5a) biosurfactants in the
organisms. The emulsification activities of each strain are not same for all four isolated strain. The
emulsification activity was measured according to the method of Cooper and Goldenberg (1987) and the result
show that APCCS1a and APCCS5a has highest emulsifying activity whereas APCCS1b lowest. The isolated strain
are tested in all experiment in triplicate manner and best represented here indicate that the natural environment
may be act as good source for alternative of toxic chemical surfactant.
5. Conclusion:
Uses of biosurfactants are increasingly in almost every sectors of the modern industry as an alternative to
chemical surfactants. With increasing public awareness in the environment, biosurfactant would most likely
replace the usage of chemical surfactants in the near future. As biosurfactants are derived from natural sources,
each of these types is an attractive alternative to synthetic compounds
References
Desai JD, Banat IM (1997) Microbial production of surfactants and their commercial
potential.MicrobiolMolBiol Rev 61: 47-64.
InkaSiegmund, Fritz Wagner (1991)New method for detecting rhamnolipids excreted by Pseudomonas species
during growth on mineral agar.Biotechnology TechniquesJul/Aug 1991, Volume 5, Issue 4, pp 265-268
B Anandaraj, P Thivakaran (2010) Isolation and production of biosurfactant producing organism from oil spilled
soil. jBiosci Tech, Vol 1 (3),2010,120‐126
T Priya, G Usharani (2009) Comparative study for biosurfactant production by using Bacillus subtilis and
Pseudomonas aeruginosa.Botany Research International 2 (4): 284-287, 2009
P. G. Carrillo, C. Mardaraz, S. I. Pitta-Alvarez, A. M. Giulietti (1996) Isolation and selection of biosurfactantproducing bacteria. World Journal of Microbiology and Biotechnology, January 1996, Volume 12, Issue 1, pp
82-84
Youssef NH, Duncan KE, Nagle DP, Savage KN, Knapp RM & McInerney MJ (2004) Comparison of methods
to detect biosurfactant production by diverse microorganisms. J Microbiol Meth 56: 339–347.
Paraszkiewicz K, KanwalA & Dlugonski J (1992) Emulsifier production by steroid transforming filamentous
fungus Curvularialunata, growth and product characterization. J Biotechnol 92: 287–294.
Morikawa M, Daido H, Takao T, Murata S, Shimonishi Y & Imanaka T (1993) A new lipopeptide biosurfactant
produced by Arthrobactersp. strain MIS 38. J Bacteriol 175: 6459–6466.
Desai
JD & Banat
IM (1997) Microbial
production
of
surfactants
and
their
commercial
potential. MicrobiolMolBiol R 61: 47–64.
Cooper DG, Goldenberg BG (1987). Surface active agents from two Bacillus species. Appl. Environ. Microbiol.,
53: 224-229.
Yin, H., Qiang, Y., Jia, J., Ye, H., Peng, H., Qin, N., Zhang, N., & He, B. (2008). Characteristics of biosurfactant
produced by Pseudomonas aeruginosa S6 isolated from oil –containing wastewater. Process Biochemistry, 44,
302-308.
OkoreChioma et.al (2013) Isolation and Characterization of Biosurfactants Producing Bacteria from Oil Polluted
Soil.Journal of Natural Sciences Research Vol.3, No.5, (119-122)
Banat I. M. (1995), Biosurfactants production and possible uses in microbial enhanced oil recovery and oil
pollution remediation: a review. Bioresource Technol. 51. 1–12.
Gautam, K. K. and Tyagi, V. K. (2005), Microbial surfactants: A review. J. Oleo. Sci., ,55, 155–166.
Rodrigues, L. (2006). Biosurfactants potential applications in medicine. J. Antimicrobial. Chemotherapy, 57,
609-618
58
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