The ability of L-asparaginase to inhibit the formation of cancer cells has aroused scientists' curiosity in biological realms. In cancer cells, L-asparaginase suppresses protein synthesis by hydrolyzing L-asparagine to L-aspartic acid and ammonia. As a result, it's a crucial therapeutic enzyme in the treatment of Acute Lymphoblastic Leukemia in combination with other drugs (ALL). This enzyme has recently been discovered to be useful in a number of scientific fields, including clinical research, pharmacology, and the food business. Purification, characterization, and assessment of the cytotoxic effect of Amycolatopsis japonica L-asparaginase were the goals of this study. Amycolatopsis japonica was isolated from the plant rhizosphere and L-asparaginase was recovered. With a molecular weight of 37.5 KDa, partially purified L-asparaginase from A. japonica had a total activity of 1968.98 U with 26.696 mg total protein and a specific activity of 73.75 U/mg, 6.42 purification fold, and 42.86 percent recovery yield. In the presence of EDTA, Mg2+, pH8, 45oC, and 0.13 mM L-asparagine, L-asparaginase from A. japonica demonstrated good activity and stability, with Km and Vmax values of 0.13 mM L-asparagine and 0.43U/ mL, respectively. The cytotoxicity of L- asparaginase from A.japonica against a colon cancer cell line was high; with an IC50 value of 36 L. Amycolatopsis japonica could be a source of L-asparaginase, which could be a new target for cancer cells.
Kinetic study of free and immobilized protease from Aspergillus sp.IOSR Journals
In the present investigation partially purified alkaline protease from Aspergillus sp. As#6 and As#7 strains were entrapped in calcium alginate beads and characterized using casein as a substrate. Temperature and pH maxima of protease from As#6 strain showed no changes before and after immobilization and remained stable at 450C and pH 9, respectively. However km value was slightly shifted from 4.5mg/ml to 5 mg/ml. Proteases from As#7 strain showed shifting in pH optima to a more alkaline range (10.0) as compared with free enzyme (9.0). Optimum temperature for protease from As#7 strain showed changes after immobilization and shifted from 650C to 850C. However there was no significant effect on Km value but Vmax of immobilized protease from As#7 strain was also shifted from 200U/ml to 370U/ml. Immobilized protease from As#6 strain was reused for 3 cycles with 22% loss in its activity whereas immobilize protease from As#7 strain was reused for 3 cycles with 17% loss in its activity. Protease from As#7 strain has a higher affinity for the substrate and higher proteolysis activity than protease from As#6 strain. The present work concludes that Aspergillus As#7 strain may be a good source of industrial protease
Impact of anthelmintic efficacy of Calotropis procera on tegumental enzymes o...iosrphr_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).
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
This is a presentation I created for my ICBI 436 Industrial Enzymology, a Biotechnology course I took at Mahidol University International College (MUIC)
Background/Purpose: The reduction solution was aqueous extracted from Acanthus ilicifolius for biosynthesis of silver nanoparticles. as green approach. It is less harmful and more economical as compared to physical and chemical methods.
Methods: Ratio of 1: 10 mixtures of 100 mg/ mL of aqueous extract and 5 mM of silver nitrate were incubated for 24 hours at 40°C with 150 rpm in incubator shaker. The formation of silver nanoparticles were monitored by colour changes and were characterized by UV-Vis spectrometry followed by zeta (potential) sizer analyses.
ABSTRACT- Aberrant glycosylation has been recognized as hallmark of cancer. Exploiting differences in glycosylation between malignant and healthy tissues offers excellent opportunities to identify sensitive and specific cancer biomarkers. Plant lectins have demonstrated the ability to specifically agglutinate malignant transformed cells. Lectins are sugar binding proteins or glycoprotein of non-immune origin which agglutinate cells or precipitate glycol-conjugates. Some lectins shown to the anti- proliferative effect on cancer cells. A wide scope of this application of lectins is that it can be used for diagnosis as well as therapeutics of cancer. The objective of the present study was to purify a lectin from tubers of Arisaema intermedium and evaluate in vitro anti-proliferative potential towards HCT-15, a human colon cancer cell line. The present study was conceived as an offshoot to the ongoing work on lectins in our laboratory. The already reported Arisaema intermedium (AIL) lectin was purified on asialofetuin linked amino-activated silica bead matrix. The purity of the affinity purified lectin was ascertained by SDS-PAGE, pH-8.3. The lectin activity was assessed by hemagglutination and protein concentration was determined by Lowry’s method. The cytotoxicity of AIL towards HCT-15 was evaluated by MTT assay. The mechanism of anti-proliferative effect was assessed by evaluation of cell morphology, trypan blue exclusion assay, DNA fragmentation and nucleic acid content determination.
Key-words- Araceae, Arisaema, Asialofetuin, Antiproliferative effect, Apoptosis, Cytotoxicity Lectins, Mechanistic
Kinetic study of free and immobilized protease from Aspergillus sp.IOSR Journals
In the present investigation partially purified alkaline protease from Aspergillus sp. As#6 and As#7 strains were entrapped in calcium alginate beads and characterized using casein as a substrate. Temperature and pH maxima of protease from As#6 strain showed no changes before and after immobilization and remained stable at 450C and pH 9, respectively. However km value was slightly shifted from 4.5mg/ml to 5 mg/ml. Proteases from As#7 strain showed shifting in pH optima to a more alkaline range (10.0) as compared with free enzyme (9.0). Optimum temperature for protease from As#7 strain showed changes after immobilization and shifted from 650C to 850C. However there was no significant effect on Km value but Vmax of immobilized protease from As#7 strain was also shifted from 200U/ml to 370U/ml. Immobilized protease from As#6 strain was reused for 3 cycles with 22% loss in its activity whereas immobilize protease from As#7 strain was reused for 3 cycles with 17% loss in its activity. Protease from As#7 strain has a higher affinity for the substrate and higher proteolysis activity than protease from As#6 strain. The present work concludes that Aspergillus As#7 strain may be a good source of industrial protease
Impact of anthelmintic efficacy of Calotropis procera on tegumental enzymes o...iosrphr_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).
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
This is a presentation I created for my ICBI 436 Industrial Enzymology, a Biotechnology course I took at Mahidol University International College (MUIC)
Background/Purpose: The reduction solution was aqueous extracted from Acanthus ilicifolius for biosynthesis of silver nanoparticles. as green approach. It is less harmful and more economical as compared to physical and chemical methods.
Methods: Ratio of 1: 10 mixtures of 100 mg/ mL of aqueous extract and 5 mM of silver nitrate were incubated for 24 hours at 40°C with 150 rpm in incubator shaker. The formation of silver nanoparticles were monitored by colour changes and were characterized by UV-Vis spectrometry followed by zeta (potential) sizer analyses.
ABSTRACT- Aberrant glycosylation has been recognized as hallmark of cancer. Exploiting differences in glycosylation between malignant and healthy tissues offers excellent opportunities to identify sensitive and specific cancer biomarkers. Plant lectins have demonstrated the ability to specifically agglutinate malignant transformed cells. Lectins are sugar binding proteins or glycoprotein of non-immune origin which agglutinate cells or precipitate glycol-conjugates. Some lectins shown to the anti- proliferative effect on cancer cells. A wide scope of this application of lectins is that it can be used for diagnosis as well as therapeutics of cancer. The objective of the present study was to purify a lectin from tubers of Arisaema intermedium and evaluate in vitro anti-proliferative potential towards HCT-15, a human colon cancer cell line. The present study was conceived as an offshoot to the ongoing work on lectins in our laboratory. The already reported Arisaema intermedium (AIL) lectin was purified on asialofetuin linked amino-activated silica bead matrix. The purity of the affinity purified lectin was ascertained by SDS-PAGE, pH-8.3. The lectin activity was assessed by hemagglutination and protein concentration was determined by Lowry’s method. The cytotoxicity of AIL towards HCT-15 was evaluated by MTT assay. The mechanism of anti-proliferative effect was assessed by evaluation of cell morphology, trypan blue exclusion assay, DNA fragmentation and nucleic acid content determination.
Key-words- Araceae, Arisaema, Asialofetuin, Antiproliferative effect, Apoptosis, Cytotoxicity Lectins, Mechanistic
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
Lipase production and purification Likhith KLIKHITHK1
Lipase (tri acyl glycerol acyl hydrolase, EC 3.1.1.3) catalyzes the hydrolysis of the carboxyl ester bonds in tri acyl glycerols to produce di acyl glycerols, mono acyl glycerols, fatty acids and glycerol under aqueous conditions and the synthesis of esters in organic solvents.
Under the controlled conditions, lipases are able to catalyze a large number of reactions. Lipases of microbial origin are of considerable commercial importance, because of the high versatility and high stability, moreover, the advantage of being readily produced in high yields.
Many microbial lipases have been commercially available in free or immobilized form. Numerous species of bacteria (Bacillus, Pseudomonas, and Burkholderia), yeasts (Candida rugosa, Yarrowia lipolytica, and Candida antarctica) and molds (Aspergillus, Trichoderma viride) produce lipases with different enzymological properties and specificities but microbes are known to be more potent lipase producer.
COMPARATIVE STUDY OF CAPSAICIN FROM IN VITRO CULTIVATED AND NATURALLY CULTIVA...Dr Dama
COMPARATIVE STUDY OF CAPSAICIN FROM IN VITRO CULTIVATED AND NATURALLY CULTIVATED CAPSICUM FRUITS EXTRACTS
*Vinchurkar A.S., *Sonawane S. R., *Sherkhane S.S., *Mane P. P., *Valsange A.B. and *Dama L. B.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
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.
In vitro mutagenesis of Cymbidium La bell “Anna Belle” by γ-rays irradiation ...IJEAB
The optimum media for multiplication of protocorm like bodies (PLBs) and shoot buds of Cymbidium La bell “Anna Belle” were studied in order to prepare the in vitro samples for irradiation. The values of LD50 (lethal dose of 50% samples) of PLBs, shoot buds and plantlets of tested Cymbidium after cultivation of 4 months were also determined about 35.0, 41.0 and 83.1 Gy, respectively. The addition of oligochitosan played as an very important trigger for promotion on the generation of shoot bud from PLBs after irradiation. The in vitro variations have been generated by γ-rays irradiation of PLBs with doses in range of 20 - 50 Gy. The highest mutant frequency (3.83‰) of C. La bell was found by the irradiation of PLB samples at 30 Gy. The different properties of obtained in vitro variations compared to wild types were found to be chlorophyll, short leaves, long leaves, and violet pericardium variations. The genetic relationships among generated variant lines in M1V4 and wild type were analyzed using RAPD techniques.
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
IJPCBS 2012, 2(1), 110-116 Kavya et al. ISSN: 2249-9504
110
INTERNATIONAL JOURNAL OF PHARMACEUTICAL, CHEMICAL AND BIOLOGICAL SCIENCES
Available online at www.ijpcbs.com
ISOLATION AND SCREENING OF STREPTOMYCES SP. FROM
CORINGA MANGROVE SOILS FOR ENZYME PRODUCTION AND
ANTIMICROBIAL ACTIVITY
M. Kavya Deepthi1*, M. Solomon Sudhakar1 and M. Nagalakshmi Devamma2 1Department of Biotechnology, Rajalakshmi Engineering College, Thandalam, 2Department of Botany, Sri Venkateswara University, Tirupati, Andhra Pr Taadmesihln, aInddui,a I.n dia.
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
Isolation Of Mung Bean (Vigna radiata (L.) R. Wilczek) Proteins To Create A S...iosrphr_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).
The Statutory Interpretation of Renewable Energy Based on Syllogism of Britis...AI Publications
The current production for energy consumption generates harmful impacts of carbon dioxide to the environment causing instability to sustainable development goals. The constitutional reforms of British Government serve to be an important means of resolving any encountered incompatibilities to political environment. This study aims to evaluate green economy using developed equation for renewable energy towards political polarization of corporate governance. The Kano Model Assessment is used to measure the equivalency of 1970 Patents Act to UK Intellectual Property tabulating the criteria for the fulfillment of sustainable development goals in respect to the environment, artificial intelligence, and dynamic dichotomy of administrative agencies and presidential restriction, as statutory interpretation development to renewable energy. The constitutional forms of British government satisfy the sustainable development goals needed to fight climate change, advocate healthy ecosystem, promote leadership of magnates, and delegate responsibilities towards green economy. The presidential partisanship must be observed to delineate parties of concerns and execute the government prescriptions in equivalence to the dichotomous relationship of technology and the environment in fulfilling the rights and privileges of all citizens. Hence, the political elites can execute corporate governance towards sustainable development of renewable energy promoting environmental parks and zero emission target of carbon dioxide discharges. The economic theory developed in statutory interpretation for renewable energy serves as a tool to reduce detrimental impacts of carbon dioxide to the environment, mitigate climate change, and produce artefacts of bioenergy and artificial intelligence promoting sustainable development. It is suggested to explore other vulnerabilities of artificial intelligence to prosper economic success.
Enhancement of Aqueous Solubility of Piroxicam Using Solvent Deposition SystemAI Publications
Piroxicam is a non-steroidal anti-inflammatory drug that is characterized by low solubility-high permeability. The present study was designed to improve the dissolution rate of piroxicam at the physiological pH's through its increased solubility by using solvent deposition system.
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Similar to Anticancer Activity of L-asparaginase Produced from Amycolatopsis japonica
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
Lipase production and purification Likhith KLIKHITHK1
Lipase (tri acyl glycerol acyl hydrolase, EC 3.1.1.3) catalyzes the hydrolysis of the carboxyl ester bonds in tri acyl glycerols to produce di acyl glycerols, mono acyl glycerols, fatty acids and glycerol under aqueous conditions and the synthesis of esters in organic solvents.
Under the controlled conditions, lipases are able to catalyze a large number of reactions. Lipases of microbial origin are of considerable commercial importance, because of the high versatility and high stability, moreover, the advantage of being readily produced in high yields.
Many microbial lipases have been commercially available in free or immobilized form. Numerous species of bacteria (Bacillus, Pseudomonas, and Burkholderia), yeasts (Candida rugosa, Yarrowia lipolytica, and Candida antarctica) and molds (Aspergillus, Trichoderma viride) produce lipases with different enzymological properties and specificities but microbes are known to be more potent lipase producer.
COMPARATIVE STUDY OF CAPSAICIN FROM IN VITRO CULTIVATED AND NATURALLY CULTIVA...Dr Dama
COMPARATIVE STUDY OF CAPSAICIN FROM IN VITRO CULTIVATED AND NATURALLY CULTIVATED CAPSICUM FRUITS EXTRACTS
*Vinchurkar A.S., *Sonawane S. R., *Sherkhane S.S., *Mane P. P., *Valsange A.B. and *Dama L. B.
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online
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.
In vitro mutagenesis of Cymbidium La bell “Anna Belle” by γ-rays irradiation ...IJEAB
The optimum media for multiplication of protocorm like bodies (PLBs) and shoot buds of Cymbidium La bell “Anna Belle” were studied in order to prepare the in vitro samples for irradiation. The values of LD50 (lethal dose of 50% samples) of PLBs, shoot buds and plantlets of tested Cymbidium after cultivation of 4 months were also determined about 35.0, 41.0 and 83.1 Gy, respectively. The addition of oligochitosan played as an very important trigger for promotion on the generation of shoot bud from PLBs after irradiation. The in vitro variations have been generated by γ-rays irradiation of PLBs with doses in range of 20 - 50 Gy. The highest mutant frequency (3.83‰) of C. La bell was found by the irradiation of PLB samples at 30 Gy. The different properties of obtained in vitro variations compared to wild types were found to be chlorophyll, short leaves, long leaves, and violet pericardium variations. The genetic relationships among generated variant lines in M1V4 and wild type were analyzed using RAPD techniques.
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
IJPCBS 2012, 2(1), 110-116 Kavya et al. ISSN: 2249-9504
110
INTERNATIONAL JOURNAL OF PHARMACEUTICAL, CHEMICAL AND BIOLOGICAL SCIENCES
Available online at www.ijpcbs.com
ISOLATION AND SCREENING OF STREPTOMYCES SP. FROM
CORINGA MANGROVE SOILS FOR ENZYME PRODUCTION AND
ANTIMICROBIAL ACTIVITY
M. Kavya Deepthi1*, M. Solomon Sudhakar1 and M. Nagalakshmi Devamma2 1Department of Biotechnology, Rajalakshmi Engineering College, Thandalam, 2Department of Botany, Sri Venkateswara University, Tirupati, Andhra Pr Taadmesihln, aInddui,a I.n dia.
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
Isolation Of Mung Bean (Vigna radiata (L.) R. Wilczek) Proteins To Create A S...iosrphr_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).
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The Statutory Interpretation of Renewable Energy Based on Syllogism of Britis...AI Publications
The current production for energy consumption generates harmful impacts of carbon dioxide to the environment causing instability to sustainable development goals. The constitutional reforms of British Government serve to be an important means of resolving any encountered incompatibilities to political environment. This study aims to evaluate green economy using developed equation for renewable energy towards political polarization of corporate governance. The Kano Model Assessment is used to measure the equivalency of 1970 Patents Act to UK Intellectual Property tabulating the criteria for the fulfillment of sustainable development goals in respect to the environment, artificial intelligence, and dynamic dichotomy of administrative agencies and presidential restriction, as statutory interpretation development to renewable energy. The constitutional forms of British government satisfy the sustainable development goals needed to fight climate change, advocate healthy ecosystem, promote leadership of magnates, and delegate responsibilities towards green economy. The presidential partisanship must be observed to delineate parties of concerns and execute the government prescriptions in equivalence to the dichotomous relationship of technology and the environment in fulfilling the rights and privileges of all citizens. Hence, the political elites can execute corporate governance towards sustainable development of renewable energy promoting environmental parks and zero emission target of carbon dioxide discharges. The economic theory developed in statutory interpretation for renewable energy serves as a tool to reduce detrimental impacts of carbon dioxide to the environment, mitigate climate change, and produce artefacts of bioenergy and artificial intelligence promoting sustainable development. It is suggested to explore other vulnerabilities of artificial intelligence to prosper economic success.
Enhancement of Aqueous Solubility of Piroxicam Using Solvent Deposition SystemAI Publications
Piroxicam is a non-steroidal anti-inflammatory drug that is characterized by low solubility-high permeability. The present study was designed to improve the dissolution rate of piroxicam at the physiological pH's through its increased solubility by using solvent deposition system.
Analysis of Value Chain of Cow Milk: The Case of Itang Special Woreda, Gambel...AI Publications
Ethiopia has a long and rich history of dairy farming, which was mostly carried out by small and marginal farmers who raised cattle, camels, goats, and sheep, among other species, for milk. Finding the Itang Special Woreda cow milk value chain is the study's main goal. In order to gather primary data, 204 smallholder dairy farmer households were randomly selected, and the market concentration ratio was calculated using 20 traders. Descriptive statistics, econometric models, and rank analysis were used to achieve the above specified goals. Out of all the participants in the milk value chain, producers, cafés, hotels, and dairy cooperatives had the largest gross marketing margins, accounting for 100% of the consumer price in channels I and II, 55% in channels III and V, and 25.5% in channels V. The number of children under five, the number of milking cows owned, the amount of money from non-dairy sources, the frequency of extension service contacts, the amount of milk produced each day, and the availability of market information were found to have an impact on smallholders' involvement in the milk market. Numerous obstacles also limited the amount of milk produced and marketed. The poll claims that general health issues, sickness, predators, and a lack of veterinary care are plaguing farmers. In order to address the issue of milk perishability, the researchers recommended the host community and organization to construct an agro milk processor, renovate the dairy cooperative in the study region, and restructure the current conventional marketing to lower the transaction and cost of milk marketing.
Minds and Machines: Impact of Emotional Intelligence on Investment Decisions ...AI Publications
In the evolving landscape of financial decision-making, this study delves into the intricate relationships among Emotional Intelligence (EI), Artificial Intelligence (AI), and Investment Decisions (ID). By scrutinizing the direct influence of human emotional intelligence on investment choices and elucidating the mediating role of AI in this process, our research seeks to unravel the complex interplay between minds and machines. Through empirical analysis, we reveal that EI not only directly impacts ID but also exerts its influence indirectly through AI-mediated pathways. The findings underscore the pivotal role of emotional awareness in investor decision-making, augmented by the technological capabilities of AI. It suggests that most investors are influenced by the identified emotional intelligence when making investment decisions. Furthermore, AI substantially impacts investors' decision-making process when it comes to investing; nevertheless, AI partially mediates the relationship between emotional intelligence and investment decisions. This nuanced understanding provides valuable insights for financial practitioners, policymakers, and researchers, emphasizing the need for holistic strategies that integrate emotional and technological dimensions in navigating the intricacies of modern investment landscapes. As the synergy between human intuition and artificial intelligence becomes increasingly integral to financial decision-making, this study contributes to the ongoing discourse on the symbiotic relationship between minds and machines in investments.0
Bronchopulmonary cancers are common cancers with a poor prognosis. It is the leading cause of death by cancer in Algeria and in the world. Behind this unfavorable prognosis hides numerous disparities according to age, sex, and exposure to risk factors, ranking 4th among incident cancers and developing countries including Algeria, all sexes combined. It ranks 2nd cancers in men and 3rd among women. Whatever the age observed, the incidence of this cancer is higher in men than in women, however the gap is narrowing to the detriment of the latter. The results of scientific research agree to relate trends in incidence and mortality rates to tobacco consumption, including passive smoking. Furthermore, other risk factors are mentioned such as exposure to asbestos in the workplace or to radon for the general population, or even genetic predisposition. However, the weight of these etiological and/or predisposing factors is in no way comparable to that of tobacco in the genesis of lung cancer and the resulting mortality. We provide a literature review in our article on the descriptive and analytical epidemiology of lung cancer.
Further analysis on Organic agriculture and organic farming in case of Thaila...AI Publications
The objective of this paper is to present Further analysis on Organic agriculture and organic farming in case of Thailand agriculture and enhancing farmer productivity. In view of the demand for organic fertilizers, efforts should also be made to enhance and to develop more effective of compost, bio-fertilizer, and bio-pesticides currently used by farmers. Likewise, emphasis should also be laid on the cultivation of legumes and other crops that can enhance the fertility of the soil, as practiced by farmers in many developing countries to fertilize their lands. On the other hand, most of the farmers who practice this farm system found that they are adopting a number of SLMs and interested in joining the meeting or training to gain more and more knowledge.
Current Changes in the Role of Agriculture and Agri-Farming Structures in Tha...AI Publications
The objective os this study is to present Current Changes in the Role of Agriculture and Agri-Farming Structures in Thailand and Vietnam with SLM practices. Farmer’s adoption and investment in SLM is a key for controlling land degradation, enhancing the well-being of society, and ensuring the optimal use of land resources for the benefit of present and future generations (World Bank, 2006; FAO, 2018). And agriculture remains an essential element of lives of many farmers in term of the strong cultural and symbolic values that attach current working generation to do and to spend time for it but not intern of income generating.
Growth, Yield and Economic Advantage of Onion (Allium cepa L.) Varieties in R...AI Publications
Haphazard and low soil fertility, low yielding verities and poor agronomic practices are among the major factors constraining onion production in the central rift valley of Ethiopia. Therefore, a field experiment was conducted in East Showa Zone of Adami Tulu Jido Combolcha district in central rift valley areas at ziway from October 2021 to April 2022 to identify appropriate rate of NPSB fertilizer and planting pattern of onion varieties. The experiment was laid out in split plot design of factorial arrangement in three replications. The main effect of NPSB blended fertilizer rates and varieties (red coach and red king) significantly (p<0.01) influenced plant height, leaf length, leaf diameter, leaf number and fresh leaf weight, shoot dry matter per plant, and harvest index. Total dry biomass, bulb diameter, neck diameter, average fresh bulb weight, bulb dry matter, marketable bulb yield, and total bulb yield were significantly (p<0.01) influenced only by the main effect of NPSB blended fertilizer rates. In addition, unmarketable bulb yield was statistically significantly affected (p≥0.05) by the blended fertilizer rates and planting pattern. Moreover, days to 90% maturity of onion was affected by the main factor of NPSB fertilizer rate, variety and planting pattern. The non-fertilized plants in the control treatment were inferior in all parameters except unmarketable bulb yield and harvest index. Significantly higher marketable bulb yield (41 t ha-1) and total bulb yield (41.33 t ha-1) was recorded from 300 kg ha-1 NPSB blended fertilizer rate applied. Double row planting method and hybrid red coach onion variety had also gave higher growth and yields. The study revealed that the highest net benefit of Birr, 878,894 with lest cost of Birr 148,006 by the combinations of 150 kg blended NPSB ha-1 with double row planting method (40cm*20cm*7cm) and red coach variety which can be recommendable for higher marketable bulb yield and economic return of hybrid onion for small scale farmers in the study area. Also, for resource full producers (investors), highest net benefit of Birr 1,205,372 with higher cost (159,628 Birr) by application of 300 kg NPSB ha-1 is recommended as a second option. However, the research should be replicated both in season and areas to more verify the recommendations.
Evaluation of In-vitro neuroprotective effect of Ethanolic extract of Canariu...AI Publications
The ethanolic extract of canarium solomonense leaves (ecsl) was studied for its neuroprotective activity. The neuroprotective activity of ECSL was found to have a significant impact on neuronal cell death triggered by hydrogen peroxide (MTT assay) in human SH-SY5Y neuroblastoma cells. Scopolamine, a muscarinic receptor blocker, is frequently used to induce cognitive impairment in laboratory animals. Injections of scopolamine influence multiple cognitive functions, including motor function, short-term memory, and attention. Using the Morris water maze, the Y maze, and the passive avoidance paradigm, memory enhancing activity in scopolamine-induced amnesic rats was evaluated. Using the Morris water maze, the Y maze, and the passive avoidance paradigm, ECSL was found to have a substantial effect on the memory of scopolamine- induced amnesic rats. Our experimental data indicated that ECSL can reverse scopolamine induced amnesia and assist with memory issues.
The goal of neuroprotection is to shield neurons against damage, whether that damage is caused by environmental factors, pathogens, or neurodegenerative illnesses. Inhibiting protein-based deposit buildup, oxidative stress, and neuroinflammation, as well as rectifying abnormalities of neurotransmitters like dopamine and acetylcholine, are some of the ways in which medicinal herbs have neuroprotective effects [1-3]. This review will focus on the ways in which medicinal herbs may protect neurons.
A phytochemical and pharmacological review on canarium solomonenseAI Publications
The genus Canarium L. consists of 75 species of aromatic trees which are found in the rainforests of tropical Asia, Africa and the Pacific. The medicinal uses, botany, chemical constituents and pharmacological activities are now reviewed. Various compounds are tabulated according to their classes their structures are given. Traditionally canarium solomonense have been used to treat a broad array of illnesses. Pharmacological actions for canarium solomonense as discussed in this review include antibacterial, antimicrobial, antioxidant, anti-inflammatory, hepatoprotective and antitumor activity.
Influences of Digital Marketing in the Buying Decisions of College Students i...AI Publications
This research investigates the influence of digital marketing channels on purchasing decisions among college students in Ramanathapuram District. The study highlights that social media marketing, online advertising, and mobile marketing exhibit substantial positive effects on purchase decisions. However, email marketing's impact appears to be more complex. Moreover, the study explores how demographic variables like gender and academic level shape these effects. Notably, freshman students display varying susceptibility to specific digital marketing messages compared to their junior, senior, or graduate counterparts. These findings offer crucial insights for marketers aiming to tailor their strategies effectively to the preferences and behaviors of college students. By understanding the differential impacts of various digital marketing channels and considering demographic nuances, marketers can refine their approaches, optimize engagement, and ultimately enhance the effectiveness of their campaigns in targeting this demographic.
A Study on Performance of the Karnataka State Cooperative Agriculture & Rural...AI Publications
The Karnataka State Co-operative Agriculture and Rural Development Bank Limited is the apex bank of all the primary co-operative agriculture and rural development banks in the state. All the PCARD Banks in the state are affiliated to it. The KSCARD Bank provides financial accommodation to the PCARD Banks for their lending operations. In order to quick sanction and disbursement of loans and supervision over the PCARD Banks the KSCARD Bank has opened district level branches. Bank has established Women Development Cell to promote entrepreneurship among women in 2005. The Bank is identifying women borrowers in the rural areas by assigning suitable projects to motivate their self-confidence to lead independent life. Progress made in financing women entrepreneurs women.
Breast hamartoma is a rare, well-circumscribed, benign lesion made up of a variable quantity of glandular, adipose and fibrous tissue. This is a lesion that can affect women at any age from puberty. With the increasingly frequent use of imaging methods such as mammography and ultrasound as well as breast biopsy, cases of hamartoma diagnosed are increasing. The diagnosis of these lesions is made by mammography. The histological and radiological aspects are variable and depend on its adipose tissue content. The identification of these lesions is important in order to avoid surgical excisions. We report radio-clinical and pathological records of breast hamartoma.
A retrospective study on ovarian cancer with a median follow-up of 36 months ...AI Publications
Ovarian cancer is relatively common but serious and has a poor prognosis. The aim of this study is to highlight the epidemiological, diagnostic, therapeutic and evolutionary aspects of this malignant pathology managed at the Bejaia university hospital center. This is a retrospective and descriptive study over a period of 3 years (2019 - 2022) carried out on 20 patients who developed ovarian cancer. The average age of the patients was 50 years old, 53.23% of whom were over 45 years old. The CA-125 blood test was positive in 18 out of 20 patients. The tumors were discovered on ultrasound in 87.10% of cases and at laparotomy in 12.90%. Total hysterectomy with bilateral adnexectomy was the most performed procedure (64.52%). The early postoperative course was simple. 15 patients underwent second look surgery (16.13%) for locoregional recurrences. Epithelial tumors were the most frequent histological type (93.55%), including 79% in the advanced stage ( IIIc -IV) and 21% in the early stage (Ia- Ib ). Adjuvant chemotherapy was administered in 80% of patients. With a median follow-up of 36 months, 2 patients were lost to follow-up. The evolution was favorable in 27.42% and in 25.81% deaths occurred late postoperatively. Ovarian cancer is not common but serious given the advanced stages and the high rate of late postoperative deaths which were largely observed in patients deprived of adequate neoadjuvant or adjuvant chemotherapy.
More analysis on environment protection and sustainable agriculture - A case ...AI Publications
This study presents a case of tea and coffee crops , esp. environment protection and sustainable agriculture in Son La and Thai Nguyen of Vietnam. Research results show us that The process of having an agricultural product goes through many steps such as planting, planning, harvesting, packing, transporting, storing and distributing. - The State adopts policies to encourage innovation of agricultural production models and methods towards sustainability, adapting to climate change, saving water, and limiting the use of inorganic fertilizers and pesticides. chemicals and products for environmental treatment in agriculture; develop environmentally friendly agricultural models. Our research limitation is that we can expand for other crops, industries and markets as well.
Assessment of Growth and Yield Performance of Twelve Different Rice Varieties...AI Publications
The present investigation entitled “Assessment of growth and yield performance of twelve different rice varieties under north Konkan coastal zone of Maharashtra” was carried out during the kharif season of the year 2021 and 2022 on the field of ASPEE, Agricultural Research and Development Foundation, Tansa Farm, At Nare, Taluka Wada, District Palghar, Maharashtra, India. The experiment was laid out in Randomized Block Design (RBD). The twelve varieties namely Zini, Jaya, Dandi, Rahghudya, Govindbhog, Dangi, Gurjari, VNR-7, VNR-8, VNR-9, Karjat-3, and Karjat-5 were replicated thrice. The plant height (cm), number of tillers per plant, number of panicles per plant, number of panicles (m²), and length of panicle (cm) were noted to the maximum with cv. “VNR-7”. The highest number of seeds per panicle, test weight (gm), grain yield (q/ha), and straw yield (q/ha) were recorded with the cv. “VNR-7”. While the lowest number of days to 50% flowering was also recorded with cv. “VNR-7” during the year 2021 and 2022.
Cultivating Proactive Cybersecurity Culture among IT Professional to Combat E...AI Publications
In the current digital landscape, cybercriminals continually evolve their techniques to execute successful attacks on businesses, thus posing a great challenge to information technology (IT) professionals. While traditional cybersecurity approaches like layered defense and reactive security have helped IT professionals cope with traditional threats, they are ineffective in dealing with evolving cyberattacks. This paper focuses on the need for a proactive cybersecurity culture among IT professionals to enable them combat evolving threats. The paper emphasis that building a proactive security approach and culture can help among IT professionals anticipate, identify, and mitigate latent threats prior to them exploiting existing vulnerabilities. This paper also points out that as IT professionals use reactive security when dealing with traditional attacks, they can use it collaboratively with proactive security to effectively protect their networks, data, and systems and avoid heavy costs of dealing with cyberattack’s aftermaths and business recovery.
The Impacts of Viral Hepatitis on Liver Enzymes and BilrubinAI Publications
Viral hepatitis is an infection that causes liver inflammation and damage. Several different viruses cause hepatitis, including hepatitis A, B, C, D, and E. The hepatitis A and E viruses typically cause acute infections. The hepatitis B, C, and D viruses can cause acute and chronic infections. Hepatitis A causes only acute infection and typically gets better without treatment after a few weeks. The hepatitis A virus spreads through contact with an infected person’s stool. Protection by getting the hepatitis A vaccine. Hepatitis E is typically an acute infection that gets better without treatment after several weeks. Some types of hepatitis E virus are spread by drinking water contaminated by an infected person’s stool. Other types are spread by eating undercooked pork or wild game. Hepatitis B can cause acute or chronic infection. Recommendation for screening for hepatitis B in pregnant women or in those with a high chance of being infected. Protection from hepatitis B by getting the hepatitis B vaccine. Hepatitis C can cause acute or chronic infection. Doctors usually recommend one-time screening of all adults ages 18 to 79 for hepatitis C. Early diagnosis and treatment can prevent liver damage. The hepatitis D virus is unusual because it can only infect those who have a hepatitis B virus infection. A coinfection occurs when both hepatitis D and hepatitis B infections at the same time. A superinfection occurs already have chronic hepatitis B and then become infected with hepatitis D. The aim of this study is to find the effect of each type of viral hepatitis on the bilirubin (TB , DSB) , and liver enzymes; AST, ALT, ALP,GGT among viral hepatitis patients. 200 patients were selected from the viral hepatitis units in the central public health laboratory in Baghdad city, all the chosen cases were confirmed as a positive samples , they are classified into four equal group each with fifty individual and with a single serological viral hepatitis type either; anti-HAV( IgM ) , HBs Ag , anti-HCV ,or anti-HEV(IgM ). All patients were tested for; serum bilirubin ( TB ,D.SB ) , AST , ALT , ALP , GGT. Another fifty quite healthy and normal person was selected as a control group for comparison. . Liver enzymes and bilirubin changes are more pronounced in HAV, HEV than HCV and HBVAST and ALT lack some sensitivity in detecting HCV ,HBV and mild elevations of ALT or AST in asymptomatic patients can be evaluated efficiently by considering ,hepatitis B, hepatitis C. ALT is generally a more sensitive indicator of acute liver cell damage than AST, It is relatively specific for hepatocyte necrosis with a marked elevations in viral hepatitis. Liver enzymes and bilirubin changes are more pronounced in HAV, HEV than HCV and HBV.AST and ALT lack some sensitivity in detecting HCV ,HBV and mild elevations of ALT or AST in asymptomatic patients can be evaluated efficiently by considering ,hepatitis B, hepatitis C. ALT is generally a more sensitive indicator of acute liver
Determinants of Women Empowerment in Bishoftu Town; Oromia Regional State of ...AI Publications
The purpose of this study was to determine the status of women's empowerment and its determinants using women's asset endowment and decision-making potential as indicators. To determine representative sample size, this study used a two-stage sampling technique, and 122 sample respondents were selected at random. To analyze the data in this study, descriptive statistics and a probit model were used. The average women's empowerment index was 0.41, indicating a relatively lower status of women's empowerment in the study area. According to the study's findings, only 40.9% of women were empowered, while the remaining 59.1% were not. The probit model results show that women's access to the media, women's income, and their husbands' education status have a significant and positive impact on the status of women's empowerment, while the family size of households has a negative impact. As a result, it is important to enhance women's access to the media and income, promote family planning and contraception, and improve men's educational status in order to improve the status of women's empowerment.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
2. Salami et al./ Anticancer Activity of L-asparaginase Produced from Amycolatopsis japonica
Int. J. Med. Phar. Drug Re. 2022 2
Vol-6, Issue-3; Online Available at: https://www.aipublications.com/ijmpd/
circulating plasma pools [6]. Meanwhile, when L-
asparaginase is present, cancer cells are unable to obtain a
critical growth factor (L-asparagine), which is required for
their continued growth, limiting their growth because they
are unable to manufacture L-asparagine in their cells. L-
asparagine is continuously reduced when this enzyme is
supplemented. The goal of this study was to generate,
purify, characterize, and test the cytotoxicity of A.
japonica L-asparaginase.
II. METHODOLOGY
2.1 Isolation of Actinomycetes
Actinomycetes A. japonica was isolated from the
rhizosphere of a plant in this investigation. Hi-media
provided the media components used in the study
(Mumbai, India). Sigma provided the asparagine
substrate Sephadex G-50 (Sigma-Aldrich, USA). All of
the substances were reagent-grade analytical compounds.
2.2 Production of L-asparaginase from Amycolatopsis
japonica
The bacterial cultured was stored on starch casein agar
(SCA) slant at 4o
C and subculture on SCA plate, and then
incubated at 27o
C for 3 days. One hundred millilitre of
M9 medium broth (Na2HPO4; 6.0 g, K2HPO4; 0.9 g,
NaCl; 0.5 g, L-asparagine; 10 g, 1M MgSO4.7H2O - 2
mL, 0.1 M solution of CaCl2.3H2O; 1m L, 20% glucose
stock; 10 mL, 0.005% phenol red, pH 6.5) in 250 mL
conical flask was inoculated with 1.5x 108
CFU/mL of 72
hrs old colony suspension and incubated at 27 °C in a
shaker incubator at 200 rpm for 7 days. At the end of the
fermentation period, the medium was centrifuged at
10,000 rpm for 15 minutes and cell-free supernatant was
taken as the crude enzyme [7].
2.3 Determination of the L-asparaginase Activity
The activity of the generated L-asparaginase enzyme was
measured using the method of Saxena et al. [7], in which
the rate of L-asparaginase hydrolysis was determined by
measuring the ammonia emitted during Nessler's reaction.
A 0.1 mL crude extract was mixed with 0.2 mL 0.05M
Tris-HCl buffer (pH 8.6) and 1.7 mL 0.01 M L-asparagine
in a 0.2 mL Tris-HCl buffer (pH 8.6). The reaction was
stopped by adding 0.5 mL of 1.5 M Trichloroacetic acid to
the mixture and incubating it for 10 minutes at 37o
C.After
centrifuging for 5 minutes at 10,000 rpm, 0.5 mL of the
supernatant was added to 7 mL distilled water and treated
with 1 mL of Nessler's reagent. The absorbance was
measured with a UV spectrophotometer at 480 nm after the
color reaction had developed for 10 minutes. It was
determined how much ammonia was liberated. The
amount of L-asparaginase enzyme that liberated 1m of
ammonia per minute under the assay conditions was
determined as one international unit [7].
𝐸𝑛𝑧𝑦𝑚𝑒 𝑎𝑐𝑡𝑖𝑣𝑖𝑡𝑦 (𝐼𝑈)
=
𝐴𝑚𝑜𝑢𝑛𝑡 𝑜𝑓 𝐴𝑚𝑚𝑜𝑛𝑖𝑎 𝐿𝑖𝑏𝑒𝑟𝑎𝑡𝑒𝑑
𝐼𝑛𝑐𝑢𝑏𝑎𝑡𝑖𝑜𝑛 𝑡𝑖𝑚𝑒 𝑥 𝑚𝐿 𝑜𝑓 𝑒𝑛𝑧𝑦𝑚𝑒 𝑢𝑠𝑒𝑑
2.4 Partial Purification of L-asparaginase
The crude enzyme was purified in three steps: ammonium
sulphate salting, dialysis desalting, and Sephadex G-50 gel
filtration chromatography [8]. Ammonium sulphate at
concentrations of 0-40 percent, 40-80 percent, and 80-100
percent (12.28 g, 14.18 g, and 7.68 g, respectively) was
dispensed into 50 mL crude enzyme in a 250 mL
Erlenmeyer flask and gently magnetically agitated until it
dissolved. These flasks were stored at 4o
C overnight before
being centrifuged for 15 minutes at 10,000 rpm at 4o
C. The
precipitate was dissolved in 0.5 M Tris-Hcl (pH 8.6) after
the supernatant was decanted [9].
Each salt concentration range's dissolved precipitate was
dialyzed. At this point, the dialysis bag was cut (5 cm
long) and soaked overnight in Tris-HCl buffer pH 8.6, 10
mL of dissolved precipitate was added to the bag, sealed
snugly, and soaked in 50 mL of the same buffer at 4 oC.
The buffer was changed every 2 hours and then left at 4 oC
overnight. The enzyme activity as well as the protein
content was measured. By putting the dialyzed enzyme on
Sephadex G-50 at 25 oC, it was further purified. After
soaking 10 g of Sephadex powder in 100 mL of Tris-Hcl
buffer pH 8.6 for 48 hours, a portion of it was put onto the
column (15cm X 2cm). The dialyzsd enzyme was loaded
on the column after it was equilibrated with the same
buffer. It was eluted at a flow rate of 0.5 mL/min using the
same buffer. The protein and enzyme activity of each
fraction was measured using the Lowry assay at 660 nm
and Nesslerisation at 480 nm on 3 mL fractions. The
fractions with the highest activity were combined, freeze-
dried, and used in further research.
2.5 Characterisation of L- asparaginase.
2.5.1 Effect of temperature on stability and activity of
L-asparaginase
The effect of temperature on the activity and stability of L-
asparaginase was investigated using the approach of
Mohamed et al. [10]. 0.1 mL of enzyme was mixed with
0.2 mL of Tris-HCl buffer pH 8.6 and 1.7 mL of 0.05 M L-
asparagine substrate in a 60-minute incubation at
temperatures ranging from 10 to 75 degrees Celsius. The
activity and stability of the enzyme were tested using the
Nesslerisation method at 10-minute intervals for 60
minutes [11].
2.5.2 Effect of pH on activity and stability of L-
asparaginase
3. Salami et al./ Anticancer Activity of L-asparaginase Produced from Amycolatopsis japonica
Int. J. Med. Phar. Drug Re. 2022 3
Vol-6, Issue-3; Online Available at: https://www.aipublications.com/ijmpd/
The activity and stability of the enzyme were tested at
various pH levels. The pH of the medium was varied
during the assay method. Different buffers (Citrate buffer;
pH4 – 5, Phosphate buffer; pH 6 – 7, Tris-Hcl buffer; pH
8, Carbonate buffer; 9 – 10) were employed to achieve a
pH range of 4 to 10. The stability and activity of the
enzyme were tested using the Nesslerisation method every
10 minutes for 60 minutes[11].
2.5.3 Influence of inducers and inhibitors on activity
and stability of L-asparaginase
This was accomplished by adding 0.1 mL of a 1 percent
inhibitor solution to the assay mixture, which included
Ascorbic acid, Urea, Triton X-100, Sodium azide, Tween
80, EDTA, and Sodium deocyl sulphate, and checking the
stability and activity of the enzyme every 10 minutes for 1
hour using the Nesslerisation method [11].
2.5.4 Influence of different metal ion on activity and
stability of L-asparaginase
To determine the effects of metal ions on the stability and
activity of the enzyme, 0.2 mL of 0.2 M of different metals
in the chloride form (Magnesium chloride, Potassium
chloride, Calcium chloride, Iron chloride, Mercury
chloride, Sodium chloride, Zinc chloride, and Ammonium
chloride) was added to the assay procedure. The activity
was monitored using the Nesslerisation method at 10-
minute intervals for an hour [12].
2.5.5 Effect of Substrate concentration on activity and
stability of L-asparaginase
The assay mixture contained 1.7 mL of various
concentrations of L- asparagine (4.0 mM, 8.0 mM, 1.2
mM, 1.6 mM, and 2.0 mM), and the effect of these
concentrations on the stability and activity of the enzyme
was determined using the Nesslerisation method at 10-
minute intervals for 1 hour [13].
2.5.6 Influence of amino acids on L-asparaginase
The enzyme's specificity towards different substrates was
determined by incubating the enzymes with 1.7 mL of 0.05
M of various amino acids such as L-aspartic acid, L-
arginine, L-phenylalanine, L-asparagines, L-glutamine,
and L-asparagine, and observing the stability and activity
using the Nesslerisation method at 10-minute intervals for
1 hour [13].
2.6 Molecular Weight Determination of L-
asparaginase.
Sodium dodecyl sulfate (SDS)-polyacrylamide gel
electrophoresis was used to evaluate the molecular weight
of the crude enzyme and to determined the purity of the
purified enzyme. 10% gel was prepared. The gel was
stained with Coomassie Brilliant Blue R-250. The protein
ladder (cat 26614) was used as reference marker. The
material used were; sample buffer( Tris-HCI (pH 6.8)
buffer 0.4 mL, 10% SDS 2.5%, 2-mercaptoethanol 0.4
mL, Glycerol 2.0 mL, Bromophenol blue 0.002g, Distilled
water 4.7 mL), Electrode buffer (Tris-HCI 6.05 g, SDS 2
g, Glycine 28.8 g, Distilled water 2.0 L), Separating (4x)
gel buffer (Tris-HCl 18.3 g, distilled water 100 mL, pH
8.8 ), Stacking (4x) gel buffer (Tris-HCl 6.055 g, distilled
water 100 mL, pH 6.8), (30%) Bisacrylamide (29.2 g
acrylamide, 0.8 g of bis-acrylamide, 100 mL).
The procedure is as follows; separating gel that comprises
of (Distilled water 19.5 mL, Bisacrylamide (30%) 10 mL,
4x separating gel buffer 10 mL , SDS (10%) 0.8 mL,
Glycerol (10%) 0.35 mL, TEMED 20 μL, APS (2%) 0.6
mL ) were mixed and gently poured in a vertical mould,
the saturated butanol was added and the gel was allowed
to polymerise. After 30 minutes, the butanol was removed
and upper portion of gel was washed with deionized water.
the stacking gel (Comprised of Distilled water 6.3 mL,
Bisacrylamide (30%) 2 mL, 4x separating gel buffer 2.5
mL, SDS (10%) 0.2 mL, Glycerol (10%) 0.15 mL,
TEMED 10 μL , APS (2%) 0.13 mL ) was poured on the
separating gel on the vertical mould and comb was placed
in it, the gel was allowed to polymerised and the comb was
removed. 30 μL of enzyme was mixed with 4X lamelli
reducing buffer to give it colour, the sample was heated at
98o
C for 10 minutes and 15 μL of sample was loaded in
the wells. Electrophoresis was carried out at 50 V until
when the dye front reached the separating gel and the
voltage was increased to 100 V. After the run is complete
the gel was taken out and washed with water. Then
commissive blue staining was carried out. The staining
solution consisted of 90 mL water, 90 mL methanol, 10
mL acetic acid and 0.25 g Commassive blue dye. While
the destaining solution consisted of 90 mL water, 90 mL
methanol, and 10 mL acetic acid. The gel was placed in
100 mL of staining solution for 30 minutes (for staining
the protein in the gel) and removed; it was then placed in
the destaining solution for destaining the gel
overnight[14].
2.7 In vitro anticancer activity of L-asparaginase from
Amycolatopsis japonica
This was done to test the activity of the crude and partially
purified enzyme on three cell lines: 3T3 (Normal cell line),
AUB5 (breast cancer cell line), and CaCo2 (intestinal
cancer cell line) (Colon cancer cell line). Cell lines were
obtained from the Panjwani Centre for Molecular
Medicine and Drug Research, ICCBS, Karachi, and were
kept in Dulbecco's Modified Eagle Medium (DMEM)
supplemented with 10% fetal bovine serum, 1% non-
essential amino acid, and cultured at 37 degrees Celsius in
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a 5% CO2 incubator. DMEM was replenished every 48-72
hours until the confluence reached 80 percent. The MTT 3-
(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium
bromide test was used to determine the cytotoxicity of
substances against the three cell lines. At a density of
10000 cells per well, 1×105
cell/mL cell suspension (100
L) was planted on a round bottom 96 well plate and
incubated at 37 oC in a 5% CO2 incubator. The cell has
grown entirely and connected to the wells after 48 hours of
incubation. The media was withdrawn, and different
concentrations of pure L-asparaginase (100, 50, and 25
µL) were combined with DMEM medium (100, 150, and
175 µL) to make a total volume of 200 L solution that was
dispensed in their respective wells and incubated for 24
hours at 37°C in a 5% CO2 incubator. Positive growth
control was provided by an untreated well. After a 24-hour
incubation period, all wells were examined under a
fluorescence microscope (Nikon Eclipse TS100 Inverted
microscope), photos were obtained, and the entire solution
was withdrawn from each well. 2 mL MTT dye (5 mg/mL)
was diluted 10 times with new DMEM media to yield 0.5
mg/mL MTT dye, and 200 L of this solution was
dispensed in each well and incubated for 2 hours at 37°C
in a 5% CO2 incubator. The MTT dye-containing media
was withdrawn after incubation, and 100 L of dimethyl
sulfoxide (DMSO) was added to all wells to solubilize the
produced formazan crystals, and absorbance was measured
in a spectrophotometer at 570 nm (Multiskan GO, Thermo
Scientific). The enzyme's cytotoxicity (percent) was
determined by comparing it to an untreated positive
growth control. The concentration that displayed 50%
cytotoxicity was calculated using a plot of percent
cytotoxicity/inhibition vs sample concentration (IC50). All
of the experiments were carried out in triplicate [15].
% cytotoxicity/inhibition = 100 -
O.D of treated well−O.D of media control
O.D of untreated control−O.D of media control
X 100
III. RESULTS AND DISCUSION
3.1 Purification
The crude L-asparaginase was purified from A.japonica
culture filtrate in three steps: ammonium sulfate
precipitation, dialysis, and Sephadex G-50 gel filtration.
The purifying procedure is depicted in (Table 1). The total
L-asparaginase activity of the crude enzyme from A.
japonica was 4593.12 U, while the total protein content
was 399.97 mg, with a specific activity of 11.48 U/mg.
The treatment with 40-80% ammonium sulphate yielded
2696.41 U with 90.71 mg total protein, 29.72 U/mg
specific activities, 2.58 purification fold, and 58.7%
recovery yield. Total activity of 2307.2 U, 33.63 mg
protein, and specific activity of 68.52 U/mg, 5.96
purification fold, and total recovery of 50.20 percent were
obtained from dialyzed 40 - 80 percent ammonium
sulphate fractions. The elution profile of the chromatogram
is shown in Figure 1, total activity of 1968.96 U, 26.69 mg
protein content, specific activity of 73.75 U/mg, 6.42 fold
purification, and recovery yield of 42.86 percent were
achieved after further purification of 40 – 80 percent
dialyzed fraction by gel filtration using Sephadex G-50.
All fractions that formed a single peak and had high L-
asparaginase activity were lyophilized.
Despite the fact that different researchers utilized nearly
identical purification methods for different enzymes, the
fold and yields of purification varied. This could be
because different proteins in the fermentation medium
have different interfaces [16].
Table 1: Summary of the purification steps of the L-asparaginase produced by Amycolatopsis japonica
Purification step Total activity
(U)
Total protein
(mg)
Specific U/Mg Purification fold % yield
Crude extract 4593.12 399.97 11.48366 1 100
Ammonium sulfate 2696.41 90.71 29.7255 2.588504 58.70
Dialysis 2307.2 33.63 68.52 5.96 50.20
Sephadex G- 50 1968.96 26.696 73.75 6.42 42.86
This finding is consistent with those published by Sahu et
al. [17], who purified enzyme from Actinomycetes strain
L9 with 18 fold and 1.9 percent recovery and a specific
activity of 13.57 U/mg. From isolated L-asparaginase from
Streptomyces tendae, Kavitha and Vijayalaksh [18] found
a specific activity of 51.7 U/mg with a purity of 17.23 fold
and a recovery of 30.5 percent. According to Narayan et
al. [19], L-asparaginase from Streptomyces albidoflavus
had a purity of 99.3 fold and a final recovery of 20%.
Lopez et al. [20] purified Streptomyces longsporusflavus
(F-15) L-asparaginase up to 30.5-fold with 19.1 percent
recovery. L-asparaginase from Streptomyces sp. PDK2 has
an 83-fold purity and 2.18 percent recovery, according to
Dhevagi and Poorani [21]. Dias et al. [22] also reported
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28.6 fold purification from A. oryzae with a yield of 6%
recovery. Kumar et al. [23] used sephadex G-100 column
chromatography to obtain 42.02 percent yield 20.91 U/mg
specific activity from L-asparaginase generated from P.
carotovorum. In addition, as each purification stage
progressed, the enzymes' particular activity rose. Amena et
al. [24] Heinen et al. [25] and Moharib [26] all report a
similar tendency.
Fig: Enzyme activity and protein content elution profile of the chromatography separation of L-asparaginase from
Amycolatopsis japonica on G-50 Sephadex column (40-80% ammonium sulphate dialyzed fraction)
3.2 Characterisation of L-asparaginase from A.
japonica
3.2.1 Effect of pH on L-asparaginase activity and
stability
The isolated A.japonica enzyme was active between pH 6
and 8, with the highest activity obtained at pH 8.0 (Figure
2). After 40 minutes of incubation at 37o
C, the enzyme
demonstrated high stability from pH (6-9) and
approximately preserved 50% of its activity in pH 8.0
(Figure 3). L-asparaginase is an amidase enzyme that has
been found to be active and stable at both alkaline and
neutral pH. At pH 8.0, the enyzme preserved 50% of its
activity for 40 minutes. Because L-asparaginase stability at
physiological pH has been reported as a determinant for
anticancer efficacy, the enzymes could be a potential
antitumor drug. Streptomyces sp. PDK7 L-asparaginase
had the highest activity at pH 8-8.5, which was similar to
this investigation [21]. At pH 7.0 and 8.0, Basha et al. [27]
found that L-asparaginase activity from marine
actinomycetes is at its peak.
Fig: Effect of pH on activity of L-asparaginase from Amycolatopsis japonica
0
2
4
6
8
10
12
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
1 3 5 7 9 111315171921232527293133353739
Protein
(mg)
Fraction number
Protein content
Enzyme activity
0
1
2
3
4
5
6
7
8
9
4 5 6 7 8 9 10
Enzyme
Activity
(U/mL)
pH range
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Fig: Effect of pH on stability of purified L–asparaginase from Amycolatopsis japonica
3.2.2 Effect of temperature on the activity and stability
of purified L-asparaginase from A. japonica.
Temperature, inferentially, is one of the decisive elements
that influence an enzyme's activity. Amycolatopsis
japonica L-asparaginase showed good activity between
35°C and 55°C, with optimal activity at 45°C. After 40
minutes of incubation, it retained between 92 and 97
percent of its activity (Figure 4 and 5). Other studies have
shown that the optimal temperature for L-asparaginase
enzyme activity is between 35o
C and 45o
C. Pseudomonas
stutzeri MB-405 and Erwinia sp. L-asparginase activity
was found to be maximum at 37°C and 35°C, respectively,
by Borkotaky and Bezbaruah, [28]. At 40°C, L-
asparaginase from Streptomyces gulbargensis had the
highest activity [29]. When L-asparaginase isolated from
Streptomyces radiopugnans MS1 was pre-incubated at
40o
C for 60 minutes, Kumar et al. [23] found no
significant loss of activity. Because of the properties of
this enzyme, it can be fully removed from the body of a
tumor patient who has received L-asparaginase in vivo.
The decrease in enzyme activity with increasing
temperature could be due to denaturation of the enzyme,
which causes a change in the active site, resulting in
inactivation of the enzyme at high temperatures.
Fig.4: Effect of temperature on activity of L-asparaginase from Amycolatopsis japonica.
0
20
40
60
80
100
120
0 10 20 30 40 50 60 70
Residual
Activity
(%)
Exposure time (Minutes)
pH 4
pH5
pH6
pH7
pH8
pH9
pH10
0
5
10
15
20
25 35 45 55 65 75
Enzyme
Activity
(U/mL)
Temperature °C
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Fig.5: Effect of temperature on stability of purified L–asparaginase from Amycolatopsis japonica.
3.2.3 Effect of various metal ion on the activity and
stability of purified L-asparaginase from A.
japonica.
The effect of various metal ions on the activity of pure L-
asparaginase was investigated. Mg2+
supported the highest
activity of pure L-asparaginase from Amycolatopsis
japonica among the metal ions examined, while Hg2+
supported the lowest activity of this enzyme (Figure 6). In
the presence of Mg2+
, the activity of Amycolatopsis
japonica enzyme was increased by 3%. In the presence of
Hg2+
, the activity of Amycolatopsis japonica L-
asparaginase was significantly reduced (Figure 7).
HgCl2 was a strong inhibitor of Streptomyces brollosae
NEAE-115 L–asparaginase, according to Noura et al. [11].
Kumar et al. [23] discovered that Hg2+
reduced nearly 80%
of the activity of Pectobacterium carotovorum L-
asparaginase. When Hg2+
and ZN2+
were present, Moharib
[30] found that V. Unguiculata L-asparaginase activity was
reduced. MgCl2 enhances the activity of Vigna radiate
purified L-asparaginase [31]. Mg2+
was found to be an
activator of L-asparaginase in Pseudomonas aeruginosa
strain SN004 in the presence of HgCl2, according to
Badoei [32].
In the presence of Hg2+
, there was enzyme inhibition
which might be due to the essential vicinal sulfhydryl
groups (SH group) of the enzyme for productive catalysis.
Mg2+
ions increase the enzyme activity suggests that these
metals ion can serve as co-factor, which can help to
activate the enzymatic reaction. Mg2+
was thought to be
the activating metal; Mg2+
may activate the substrate,
bound directly to the enzyme-substrate complex.
Mg2+
locks the enzyme-substrate complex in place and
then rapidly causes release of the reaction products [33].
This corresponds to fast dissociation rates for the enzyme-
product complex rendering more favorable substrate
binding sites. Metal ions play a crucial role in maintaining
the active configuration of the enzymes at elevated
temperatures by protecting them against thermal
denaturation [34].
Fig: Effect of metal ions on activity of L-asparaginase from Amycolatopsis japonica
0
20
40
60
80
100
120
140
160
180
0 20 40 60 80
Residual
Activity
(%)
Exoposure time (minutes)
25°C
35°C
45°C
55°C
65°C
0
2
4
6
8
10
12
14
16
Ca²⁺ Fe²⁺ K⁺ Mg²⁺ Na⁺ Zn²⁺ Hg²⁺
Enzyme
Activity
(U/mL)
Metal Ions
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Fig.7: Effect of metal ions on stability of purified L–asparaginase from Amycolatopsis japonica
3.2.4 Effect of Inhibitors and Inducers on the activity
and stability of purified L-asparaginase from A.
japonica.
EDTA acted as inducers for the purified L-asparaginase
from Amycolatopsis japonica with maximum activity of
18.73 U/mL, the lowest activity was found in the presence
of ascorbic acid with enzyme activity of 4.89 U/mL. The
enzyme showed good stability within 60 minutes of
incubation and retained up to 60% of its activity in the
presence of EDTA. This result obtained is in line with the
work of Dias et al. [22] who reported slight decrease of
around 30% of L–asparaginase from Aspergillus oryzae in
the presence of EDTA. Noura et al. [11] recorded that
Tween 80 acted as activators for L-asparaginase activity
from Streptomyces brollosae NEAE-115 and reported
EDTA to be an inhibitor of L–asparaginase
from Streptomyces brollosae NEAE-115 as slight decrease
of about 37.55% in the activity of the enzyme was
observed. On the contrary, Moorthy et al. [35] and
Jayachandra, [31] recorded inhibition of L-asparaginase
activity from Bacillus sp. and Vigna radiate respectively
by EDTA.
The fact that the activity of the enzyme was inhibited in
the presence of some metal ion and was not inhibited in the
presence of EDTA shows that the enzyme was not a
metalloprotein. Direct and quick contact of enzyme with
substrate sites seems to be increase by biosurfactant, and
this might be why Tween 80 supported the activity of the
enzyme [36]. Tween 80 enhanced substrate binding
capacity and stability of enzymes under in vitro conditions
[37]
Fig.8: Effect of Inhibitors and Inducers on activity of L-asparaginase from Amycolatopsis japonica
0
20
40
60
80
100
120
10 20 30 40 50 60
Residual
Activity
(%)
Metal ions
Ca²⁺
Fe²⁺
K⁺
Mg²⁺
Na⁺
Zn²⁺
0
2
4
6
8
10
12
14
16
18
20
Triton-XAscorbic
acid
Urea Sodun
azide
Tween
80
EDTA SDS
Enzyme
Activity
(U/mL)
Inducers and Inhibitors
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Fig.9: Effect of inhibitors and inducers on stability of purified L–asparaginase from Amycolatopsis japonica
3.2.5 Effect of amino acids on the activity and stability
of purified L-asparaginase from A. japonica.
The purified L-asparaginase from Amycolatopsis japonica
showed highest activity when L-asparagine was present
with lowest activity in the presence of L-glutamine, and
retained 60% of its activity in the presence of L-asparagine
after 50 minutes of incubation (Figure 10 and 11)
The enzyme production is the complex chain reactions
and is supported and induced by suitable substrates [38].
One of the properties of enzymes that make them useful
as diagnostic tools is their specificity towards their
substrate. The enzyme showed high specificity towards
its natural substrate L-asparagine, very low specificity
towards L-aspartic acid, while no activity towards L-
glutamine.
This is in line with [38] who reported that Penicillium sp.
preferred L–asparagine as substrate and contrary to
Dunlope and Roon [39] who noted the increment in L-
asparaginase production from Penicillium sp due to the
addition of L glutamine or glutamate in the fermentation
medium.
Fig.10: Effect of amino acids on activity of L-asparaginase from Amycolaptosis japonica from rhizospheric soil.
0
20
40
60
80
100
120
0 10 20 30 40 50 60 70
Residual
Activity
(%)
Exposure time
Triton-X
Ascorbic acid
Tween 80
EDTA
SDS
Sodun azide
Urea
0
5
10
15
20
25
30
35
40
Enzyme
Activity
(U/mL)
Amino acids
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Fig.11: Effect of amino acids on stability of purified L–asparaginase from Amycolatopsis japonica
3.2.6 Effect of Substrate concentrations on the
activity and stability of purified L-asparaginase
from A. japonica.
L-asparaginase of Amycolatopsis japonica had Vmax
value of 0.13 mM and Km value of 0.43 U/mL (Figure 12
and 13). Kotzia and Labrou, (2007) reported that Erwinia
chrysanthemi L-asparginase had Km value of (0.058 mM).
Km and Vmax of purified L-asparaginase from F. culmorum
ASP-87 were reported to be 3.1 mM and 0.77 μmol/
ml/min respectively. Km value of P. brevicompactum L-
asparaginase was reported by Elshafei et al. ([41] to be
1.05 mM. Dias et al. [22] reported that L-asparaginase
from A. oryzae CCT 3940 demonstrated high affinity for
the substrate L-asparagine with Km and Vmax values
estimated in 0.66 mol/L and 313 U/mL, respectively. Km
value of 7.14 mM was recorded for L-asparaginase from
Erwinia carotovora by Kamble et al. [42]. Higher Km
values 6.6 and 7.0 mM for L-asparaginase from Lupinus
arboreus and Lupinus angustifolius, respectively, has been
reported [43]. However, slightly higher Km values of 12.5
mM were reported in Aspergillus aculeatus [44].
Summarily, from the above values, there exist great
variations in the kinetic parameters of L-asparaginase from
different sources. However, substrate concentration is
another environmental factor that affects enzymes activity.
The rate of an enzyme-catalysed reaction increases with
increase in substrates concentration. Therefore the
maximum velocity (Vmax) and Michaelis Constant (Km)
of enzymes are usually determine in order to know the best
substrates loading for enzymes meant to be applied. Vmax
refers to the rate at which an enzyme converts its
substrates to products in the presence of excess substrates,
while Km refers to the concentration of substrates at which
an enzyme acts at half its maximum velocity. Km is also a
measure of the apparent substrates affinity of an enzyme.
From this study, the enzyme has high affinity to it natural
substrate, which can be reasons for its degree of inhibition
against cancer cell lines. Raha et al. [45] noted that the
effectiveness of an L- asparaginase enzyme against tumor
is dependent on its affinity to its substrate.
Fig.12: Effect of substrate concentration on activity of L – asparaginase from Amycolatopsis japonica
0
20
40
60
80
100
120
0 10 20 30 40 50 60 70
Residual
Activity
(%)
Exposure time (Minutes)
L-aspartic acid
L-asparagine
L-phenylalanine
L-glutamine
L-arginine
0
1
2
3
4
0.4 0.8 1.2 1.6 2
Enzyme
Activity
(U/mL)
Substrate Concentrations (mM)
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Fig.13: Line weaver - burk plot for the reaction kinetics of L-asparaginase from Amycolatopsis japonica
Cytotoxicity
3.3 In vitro anticancer activity of L-asparaginase
produced.
L-asparaginase from Amycolatopsis japonica showed more
inhibitory activity against colon cancer cell line (Caco-2)
and breast cancer cell line (AU5) but less on normal rat
fibroblast (3T3) cell line (Figure 14). The enzyme
exhibited more effectiveness on growth inhibition on colon
cancer cell line but less on breast cancer cell lines. The
incubation of colon cancer cell line with gradual doses of
L-asparaginase from this organisms led to a gradual
inhibition in the cell growth with IC50 values of 36μL
(Figure15)
Moharib, [30] recorded that, L-asparaginase from Vigna
unguiculata seed have more anticancer effect against liver
(HEPG2) and colon (HCT-116) but lower effective against
cervical (HELA) and Breast (MCF7) cancer cell lines.
Studied the in vitro cytotoxicity of Bacillus sp R36 L-
asparaginase against different cell lines and reported that,
the enzyme have more cytotoxic effect on liver cancer cell
line (Hep G2) than colon cancer cell line (HCT- 116) [46].
Dias et al., 2016 studied cytotoxic activity of L-
asparaginase produced from purified L-asparaginase from
A. oryzae CCT 3940 on broad range of human tumor cell
lines (786-0 (kidney), NCI-H40 (lung, non-small cell), PC-
3 (prostate),Type, U251 (glioma), UACC-62 (melanoma),
HT29 (colon) and K562 (leukemia) at different
concentration and reported that the enzyme completely
inhibited the cell proliferation of these cell lines and did
not inhibit the non-carcinogenic human cell line growth at
the concentrations studied.
The antineoplastic activity of the L-asparaginase produced
by the isolated bacterial was performed based on the fact
that lymphatic cells demand huge quantities of L-
asparagine in order to have rapid malignant growth as
these tumor cells lack or have very low expression levels
of L-asparagine synthetase and depend on the extracellular
pool of this amino acid unlike normal cells. The presence
of an external L-asparaginase enzyme in the growth
medium causes depletion of asparagine due to the catalysis
of the supplemented enzyme and kills tumor cells by
depriving them of an essential factor required for protein
synthesis [47].
Tumor cells are destroyed by L-asparaginase without
significant damage to normal cells. This explains less
inhibitory activity of this enzyme on 3T3 rat fibroblast cell
because cancer cells are L-asparagine dependent, an amino
acid essential for lymphoblasts growth. Non-cancerous cell
has the ability to manufacture L-asparagine and cannot be
affected by L-asparaginase treatment, because they contain
L-asparagine synthase, cancerous cells do not have L-
asparaginase synthase, and so they cannot produce L-
asparaginase on their own, thereby affected when treated
with L-asparaginase [40].
Km=0.13 mM
Vmax=0.43 mL
y = 0.352x + 2.631
R² = 0.942
0
1
2
3
4
5
6
-8 -6 -4 -2 0 2 4 6 8
12. Salami et al./ Anticancer Activity of L-asparaginase Produced from Amycolatopsis japonica
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Fig.14: Cytotoxicity effect of L-asparaginase from Amycolatopsis japonica on three cell lines
Keys:
3t3: Rat fibroblast cell line
AUB5: Breast Cancer cell line
CACO2: Colon cancer cell line
Plate 1: Microscopic picture of anticancer activity of L-asparaginase from Amycolatopsis japonica on colon cancer cell line
at different concentrations.
Keys: (a) 100 μL; 83% dead cells, (b)50 μL; 78% dead cells with few live ones, (c) 25 μL; Live cells with 26% dead cells,
(d) Control; Lives cells
0
10
20
30
40
50
60
70
3t3 AUB5 CACO
Growth
inhibition
(%)
Cancer Cell lines
13. Salami et al./ Anticancer Activity of L-asparaginase Produced from Amycolatopsis japonica
Int. J. Med. Phar. Drug Re. 2022 13
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Fig.15: Cytotoxicity effect of different concentration of L-asparaginase from Amycolatopsis japonica on Colon cancer cell
line
IV. CONCLUSION
From this result, it could be concluded that L-asparaginase
from Amycolatopsis japonica exhibited anticancer
potential and could be used as drug to complement the
ones currently in use. To the best of my knowledge, this is
the first report on anticancer activity of L-asparaginase
from Amycolatopsis japonica.
ACKNOWLEDGEMENT
My heartfelt thanks go to the World Academy of Science
Scholarship board for their financial support in carrying
out this research. Also, thanks to the International Center
for Chemical and Biological Sciences, University of
Karachi, Pakistan for providing the space and materials for
this work, and to the Panjwani Centre for Molecular
Medicine and Drug Research, ICCBS, Karachi, Pakistan
for providing the cell lines used in this study.
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