THERAPEUTIC L-ASPARAGINASE ACTIVITY OF BACTERIA ISOLATED FROM MARINE SEDIMENTS
Abstract
L-Asparaginase, a therapeutic enzyme used in lymphoblastic leukemia and lymphosarcoma chemotherapy which is derived mostly from the bacterial sources Escherichia coli and Erwinia sp. The long term administration of the drug leads to the development of resistant tumours and anaphylactic shock in certain individuals. Hence serologically different L-Asparaginase from novel microbial sources with enhanced therapeutic potential and immunological characteristics is an essential requirement. The marine bacteria having diverse range of potential enzymes might be a source for L-Asparaginase with novel properties, which are still unexplored. In this study, we have screened marine bacteria isolated from the coastal regions of Kerala which showed both intra and extra cellular L-Asparaginase activity. Bacillus sp. (Accession no KF142395) was found to have the highest extracellular enzyme activity (2.31 IU/ml) while Shewanella sp. (Accession no KF142390) showed maximum intracellular Asparaginase activity (2.16 IU/ml).The crude extracellular enzyme preparation from Bacillus sp. had cytotoxic effect on HL60 cell line with an IC50 value of 12.5µg/ml.
Keywords:
L-Asparaginase, 16S rRNA, Antineoplastic, Therapeutic-enzyme, MTT assayDOI
https://doi.org/10.25004/IJPSDR.2016.080407References
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2. Gulati R, Saxena RK, Gupta R. A rapid plate assay for screening L-Asparaginase producing micro-organisms. Lett Appl Microbiol. 1997; 24: 23-26.
3. Kidd JG. Regression of transplanted lymphomas induced in vivo by means of normal guinea pig serum. I. Course of transplanted cancers of various kinds in mice and rats given guinea pig serum, horse serum, or rabbit serum. J Exp Med. 1953; 98: 565–582.
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5. Pedreschi F, Kaack K, Granby K. The effect of Asparaginase on Acrylamide formation in french fries. Food Chem. 2008; 109:386-392.
6. Shah AJ, Karadi RV, Parekh PP. Isolation, Optimization and production of L- Asparaginase from coliform bacteria. Asian J Biotechnol. 2010; 2:169-177.
7. Ahmad N, Pandit NP, Maheshwari SK. L-Asparaginase gene - a therapeutic approach towards drugs for cancer cell. Int J Biosci. 2012; 2:1-11.
8. Cedar H, Schwartz JH. Production of L-Asparaginase II by Escherichia coli. J Bacterio. 1968; 96: 2043-2048.
9. Deokar VD, Vetal MD, Rodrigues L. Production of intracellular L- Asparaginase from Erwinia carotovora and its statistical optimization using response surface methodology (RSM). Int J Chem Sci Appl. 2010; 1: 25-26.
10. Tosa T, Sano R, Yamamoto K, Nakamura M, Ando K, Chibata I. L-Asparaginase from Proteus vulgaris purification, crystallization and enzymic properties. Biochem. 1972; 11:217–222.
11. Sukumaran CP, Singh DV, Mahadevan PR. Synthesis of L-Asparaginase by Serratia marcescens (Nima). J Biosci. 1979; 1:263–269.
12. Mukherjee J, Majumdar S, Scheper T. Studies on nutritional and oxygen requirements for production of L-Asparaginase by Enterobacter aerogenes. Appl Microbiol Biotechnol 2000; 53:180-184.
13. Prakasham RS, Rao C, Rao RS, Lakshmi GS, Sarma PN. L‐Asparaginase production by isolated Staphylococcus sp.–6A: design of experiment considering interaction effect for process parameter optimization. J Appl Microbiol. 2007; 102:1382-1391.
14. Abdel-Fattaha YR, Olama ZA. L-Asparaginase production by Pseudomonas aeruginosa in solid-state culture: evaluation and optimization of culture conditions using factorial designs. Process Biochem. 2002; 38:115-122.
15. Pritsa AA, Kyriakidis DA. L-Asparaginase of Thermus thermophiles: purification, properties and identification of essential amino acids for catalytic activity. Mol Cell Biochem. 2001; 216: 93-101.
16. Maysa EM, Amira M, Gamal E, Sanna T, Sayed EI. Production, immobilization and anti-tumor activity of L-Asparaginase of Bacillus sp. R36. J Am Science. 2010; 6: 157-165.
17. Amena S, Vishalakshi N, Prabhakar M, Dayanand A, Lingappa K. Production, purification and characterization of L-Asparaginase from Streptomyces gulbargensis. Braz J Microbiol. 2010. 41: 173-178.
18. Dejong PJ. L-Asparaginase production by Streptomyces griseus. Appl Microbiol. 1972; 23: 1163-1164.
19. Avarmis VI, Tiwari PN. Asparaginase (native ASNase pegylatedASNase) in the treatment of acute lymhoblastic leukemia. Int J Nanomedicine. 2006; 1: 241-254.
20. Eden OB, Shaw MP, Lilleyman JS, Richards S. Non-randomised study comparing toxicity of Escherichia coli and Erwinia Asparaginase in children with leukemia. Med Pediatr Oncol. 1990; 18:497–502.
21. Wriston JC, Yellin TO. “L-Asparaginase: A Review”. In: Meister A, editor. Advances in Enzymology and Related Areas of Molecular Biology. John Wiley & Sons, Inc., Hoboken, NJ, USA. 1973; 39:185-248.
22. Polz MF, Harbison C, Cavanaugh CM. Diversity and heterogeneity of epibiotic bacterial communities on the marine nematode Eubostricus dianae. Appl Environ Microbiol. 1999; 65:4271-4275.
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25. Ghasemi Y, Ebrahiminezhad A, Rasoul-Amini S, Zarrini G, Ghoshoon MB, Raee MJ et al. An optimized medium for screening of L-Asparaginase production by Escherichia coli. Am J Biochem Biotechnol. 2008; 4:422-424.
26. Zhang L, Foxman B, Gilsdorf JR, Marrs CF. Bacterial genomic DNA isolation using sonication for microarray analysis. Biotechniques. 2005; 39: 640–644.
27. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65:55-63.
28. Ebrahiminezhad A, Rasoul-Amini S, Ghasemi Y. L-Asparaginase production by moderate halophilic bacteria isolated from Maharloo salt lake. Indian J Microbiol. 2011; 51: 307–311.
29. Rani SA, Lalitha S, Vasantha PB. In-vitro antioxidant and anticancer activity of L-Asparaginase from Aspergillus flavus (KUFS20). Asian J Pharm Clin Res. 2011; 4:174-177.
30. Selvam K, Vishnupriya B. Partial purification and cytotoxicity activity of L-Asparaginase from Streptomyces acrimycini NGP. Int J Res Pharm Biomed Sci. 2013; 4:859-869.
31. Sabu A. Sources, properties and applications of microbial therapeutic enzymes. Indian J Biotechnol. 2003; 2:334–341
32. Imada A, Igarasi S, Nakahama K, Isono M. Asparaginase and glutaminase activities of micro-organisms. J Gen Microbiol. 1973; 76: 85-99.
33. Prakasham RS, Hymavathi M, Rao CS, Arepalli SK, Rao JV, Kennady PK, et al. Evaluation of antineoplastic activity of extracellular Asparaginase produced by isolated Bacillus circulans. Appl Biochem Biotechnol. 2010; 160:72-80.
34. Moorthy V, Ramalingam A, Sumantha A, Shankaranaya RT. Production, purification and characterization of extracellular L-Asparaginase from a soil isolate of Bacillus sp. Afr J Microbiol Res. 2010; 4:1862-1867.
35. Ravenschlag K, Sahm K, Pernthaler J, Amann R. High bacterial diversity in permanently cold marine sediments. Appl Environ Microbiol. 1999; 65:3982-3989.
36. Stach JE, Maldonado LA, Masson DG, Ward AC, Goodfellow M, Bull AT. Statistical approaches for estimating actinobacteria diversity in marine sediments. Appl Environ Microbiol. 2003; 69:6189-6200.
2. Gulati R, Saxena RK, Gupta R. A rapid plate assay for screening L-Asparaginase producing micro-organisms. Lett Appl Microbiol. 1997; 24: 23-26.
3. Kidd JG. Regression of transplanted lymphomas induced in vivo by means of normal guinea pig serum. I. Course of transplanted cancers of various kinds in mice and rats given guinea pig serum, horse serum, or rabbit serum. J Exp Med. 1953; 98: 565–582.
4. Broome JD. Evidence that the L - Asparaginase activity of guinea pig serum is responsible for its antilymphoma effects I. Properties of the L-Asparaginase of Guinea pig serum in relation to those of the antilymphoma substance. J Exp Med. 1963; 118: 99–120.
5. Pedreschi F, Kaack K, Granby K. The effect of Asparaginase on Acrylamide formation in french fries. Food Chem. 2008; 109:386-392.
6. Shah AJ, Karadi RV, Parekh PP. Isolation, Optimization and production of L- Asparaginase from coliform bacteria. Asian J Biotechnol. 2010; 2:169-177.
7. Ahmad N, Pandit NP, Maheshwari SK. L-Asparaginase gene - a therapeutic approach towards drugs for cancer cell. Int J Biosci. 2012; 2:1-11.
8. Cedar H, Schwartz JH. Production of L-Asparaginase II by Escherichia coli. J Bacterio. 1968; 96: 2043-2048.
9. Deokar VD, Vetal MD, Rodrigues L. Production of intracellular L- Asparaginase from Erwinia carotovora and its statistical optimization using response surface methodology (RSM). Int J Chem Sci Appl. 2010; 1: 25-26.
10. Tosa T, Sano R, Yamamoto K, Nakamura M, Ando K, Chibata I. L-Asparaginase from Proteus vulgaris purification, crystallization and enzymic properties. Biochem. 1972; 11:217–222.
11. Sukumaran CP, Singh DV, Mahadevan PR. Synthesis of L-Asparaginase by Serratia marcescens (Nima). J Biosci. 1979; 1:263–269.
12. Mukherjee J, Majumdar S, Scheper T. Studies on nutritional and oxygen requirements for production of L-Asparaginase by Enterobacter aerogenes. Appl Microbiol Biotechnol 2000; 53:180-184.
13. Prakasham RS, Rao C, Rao RS, Lakshmi GS, Sarma PN. L‐Asparaginase production by isolated Staphylococcus sp.–6A: design of experiment considering interaction effect for process parameter optimization. J Appl Microbiol. 2007; 102:1382-1391.
14. Abdel-Fattaha YR, Olama ZA. L-Asparaginase production by Pseudomonas aeruginosa in solid-state culture: evaluation and optimization of culture conditions using factorial designs. Process Biochem. 2002; 38:115-122.
15. Pritsa AA, Kyriakidis DA. L-Asparaginase of Thermus thermophiles: purification, properties and identification of essential amino acids for catalytic activity. Mol Cell Biochem. 2001; 216: 93-101.
16. Maysa EM, Amira M, Gamal E, Sanna T, Sayed EI. Production, immobilization and anti-tumor activity of L-Asparaginase of Bacillus sp. R36. J Am Science. 2010; 6: 157-165.
17. Amena S, Vishalakshi N, Prabhakar M, Dayanand A, Lingappa K. Production, purification and characterization of L-Asparaginase from Streptomyces gulbargensis. Braz J Microbiol. 2010. 41: 173-178.
18. Dejong PJ. L-Asparaginase production by Streptomyces griseus. Appl Microbiol. 1972; 23: 1163-1164.
19. Avarmis VI, Tiwari PN. Asparaginase (native ASNase pegylatedASNase) in the treatment of acute lymhoblastic leukemia. Int J Nanomedicine. 2006; 1: 241-254.
20. Eden OB, Shaw MP, Lilleyman JS, Richards S. Non-randomised study comparing toxicity of Escherichia coli and Erwinia Asparaginase in children with leukemia. Med Pediatr Oncol. 1990; 18:497–502.
21. Wriston JC, Yellin TO. “L-Asparaginase: A Review”. In: Meister A, editor. Advances in Enzymology and Related Areas of Molecular Biology. John Wiley & Sons, Inc., Hoboken, NJ, USA. 1973; 39:185-248.
22. Polz MF, Harbison C, Cavanaugh CM. Diversity and heterogeneity of epibiotic bacterial communities on the marine nematode Eubostricus dianae. Appl Environ Microbiol. 1999; 65:4271-4275.
23. Hall T. BioEdit version 5.0.6, Department of Microbiology, North Carolina State University North Carolina State University. 2001; http:// www.mbio.ncsu.edu/BioEdit/bioedit.html.
24. Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) Software Version 4.0. Mol Biol Evol. 2007; 24:1596-1599.
25. Ghasemi Y, Ebrahiminezhad A, Rasoul-Amini S, Zarrini G, Ghoshoon MB, Raee MJ et al. An optimized medium for screening of L-Asparaginase production by Escherichia coli. Am J Biochem Biotechnol. 2008; 4:422-424.
26. Zhang L, Foxman B, Gilsdorf JR, Marrs CF. Bacterial genomic DNA isolation using sonication for microarray analysis. Biotechniques. 2005; 39: 640–644.
27. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65:55-63.
28. Ebrahiminezhad A, Rasoul-Amini S, Ghasemi Y. L-Asparaginase production by moderate halophilic bacteria isolated from Maharloo salt lake. Indian J Microbiol. 2011; 51: 307–311.
29. Rani SA, Lalitha S, Vasantha PB. In-vitro antioxidant and anticancer activity of L-Asparaginase from Aspergillus flavus (KUFS20). Asian J Pharm Clin Res. 2011; 4:174-177.
30. Selvam K, Vishnupriya B. Partial purification and cytotoxicity activity of L-Asparaginase from Streptomyces acrimycini NGP. Int J Res Pharm Biomed Sci. 2013; 4:859-869.
31. Sabu A. Sources, properties and applications of microbial therapeutic enzymes. Indian J Biotechnol. 2003; 2:334–341
32. Imada A, Igarasi S, Nakahama K, Isono M. Asparaginase and glutaminase activities of micro-organisms. J Gen Microbiol. 1973; 76: 85-99.
33. Prakasham RS, Hymavathi M, Rao CS, Arepalli SK, Rao JV, Kennady PK, et al. Evaluation of antineoplastic activity of extracellular Asparaginase produced by isolated Bacillus circulans. Appl Biochem Biotechnol. 2010; 160:72-80.
34. Moorthy V, Ramalingam A, Sumantha A, Shankaranaya RT. Production, purification and characterization of extracellular L-Asparaginase from a soil isolate of Bacillus sp. Afr J Microbiol Res. 2010; 4:1862-1867.
35. Ravenschlag K, Sahm K, Pernthaler J, Amann R. High bacterial diversity in permanently cold marine sediments. Appl Environ Microbiol. 1999; 65:3982-3989.
36. Stach JE, Maldonado LA, Masson DG, Ward AC, Goodfellow M, Bull AT. Statistical approaches for estimating actinobacteria diversity in marine sediments. Appl Environ Microbiol. 2003; 69:6189-6200.
Published
01-07-2016
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“THERAPEUTIC L-ASPARAGINASE ACTIVITY OF BACTERIA ISOLATED FROM MARINE SEDIMENTS”. International Journal of Pharmaceutical Sciences and Drug Research, vol. 8, no. 4, July 2016, pp. 229-34, https://doi.org/10.25004/IJPSDR.2016.080407.
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How to Cite
“THERAPEUTIC L-ASPARAGINASE ACTIVITY OF BACTERIA ISOLATED FROM MARINE SEDIMENTS”. International Journal of Pharmaceutical Sciences and Drug Research, vol. 8, no. 4, July 2016, pp. 229-34, https://doi.org/10.25004/IJPSDR.2016.080407.
