Production and Characterization of Partially Purified Chitosanase Enzyme of Bacillus cereus A4/B4 Isolated from Biowaste Soil Samples:Bioactive Chitooligosaccharide

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Authors

  • P. Vanathi
     Department of Microbiology, Maharaja Co Education Arts & Science College, Perundurai ,IN
  • S. Fernandez
     Department of Microbiology, Maharaja Co Education Arts & Science College, Perundurai ,IN
  • M. Shanmugapriya
     Department of Microbiology, Maharaja Co Education Arts & Science College, Perundurai ,IN
  • T. S. Riyamol
     Department of Microbiology, Maharaja Co Education Arts & Science College, Perundurai ,IN
  • V. Loganathan
     Department of Microbiology, Maharaja Co Education Arts & Science College, Perundurai ,IN
  • V. I. Hairul-Islam
     Division of Microbiology and immunology, Pondicherry centre for biological sciences, Pondicherry ,IN
  • P. Premasudha
     Department of Nanotechnology, Bharathiar University, Coimbatore ,IN
  • R. Rajendran
     PSG College of Arts & Science, Coimbatore ,IN

DOI:

https://doi.org/10.15613/fijrfn/2015/v2i2/96643

Keywords:

Bacillus Species, Bioproduction, Chitosanase, Chitooligosaccharides, Microbial Enzymes.

Abstract

Microbial chitosanase has received the attention of producing chitooligosaccharides. In this study, Chitooligosaccharide was developed by chitosanase enzyme hydrolysis and evaluated for its bioactivity. Chitosan degrading Bacillus cereus was isolated from biowaste sample and its extracellular enzyme chitosanase was characterized. The chitosanase enzyme production was induced by chitosan substrate and the enzyme production reached the maximum in 72 hours. The optimum pH and temperature for the production was 6.5 and 30°C respectively. The culture medium with 1.0 to 1.5% of substrate level with xylose served as significantly for enzyme production. The molecular weight of the enzyme 43KDa was determined by SDS-PAGE. The chitooligosaccharide produced by chitosanase hydrolysis inhibited the growth of the pathogens and showed MIC50 as 0.2 μg/ml against all the test pathogens and has proved to be a potential antimicrobial agent against skin and other infections.

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Published

2015-12-01

Issue

Volume 2, Issue 2, July-December 2015

Section

Articles

 

References

Woo–Jin J., Ro–Dong P. “Bioproduction of Chitooligosaccharides: Present and Perspectivesâ€, Marine Drugs, vol. 12, p. 5328–5356, 2014.

Xu- Fen Z., Hai-Qin T., Chu Z., Li L., Xin-Qi Z., and Min W. “Cloning and overexpression of a new chitosanase gene from Penicillium sp.D-1â€, AMB Express(Springer Open Journal), vol. 2, p. 13, 2012.

Farooqahamed S. K., Vishu K. A. B., Lalitha R. G., Rudrapatnam N. T. “Chitosanolysis by a pectinase isozyme of Aspergillus niger –A non specific activityâ€, Carbohydrate polymer, vol. 53, p. 191–196.

Mei-Chin C., Fwu-Long M., Zi-Xian L., and Hsing-Wen S. “Chitosan:Its applications in Drug eluting Devicesâ€, Advances in Polymer Science, vol. 243, p. 185–230, 2011.

Ho-Geun Y., Hee-Yun K., Hye-Kyung K., Bum-Shik H., Dong-Hoon S., and Hong –Yon C. “Thermostable chitosanae from Bacillus sp.Strain CK4:its Purification,Characterization and Reaction Patternsâ€, Bioscience Biotechnology Biochemistry, vol. 65(4), p. 802–809.

Cheng C. Y., and Li Y. K. “An Aspergillus chitosanse with potential for large scale preparation of Chitosan oligosaccharidesâ€, Biotechnology and Applied Biochemistry, vol. 32, p. 197–203, 2000.

Cheng S. W., Wang Y. F., and Hong B. “Statistical optimization of medium compositions for chitosanase production for chitosanase production by a newly isolated Streptomyces albusâ€, Brazilian Journal of Chemical Engineering, vol. 29(4), p. 691–698, 2012.

Park J. K., Kumiko S., Nobuhisa O., Kazutaka S., Masako K., Yukari O., Katsunori T., Hideyuki M., and Makoto K. “Purification, characterization and gene analysis of a chitosanase (ChoA) from Matsuebacter chitosanotabidus 3001â€, Journal of Bacteriology, vol. 181(21), p. 6642–6649, 1999.

Yuying S., Jiquan Z., Shujun W. “Purification and characterization of the chitosanase from Aeromonas sp.HG08â€, African Journal of Biotechnology, vol. 8(12), p. 2830–2834, 2009.

Choi Y. J., Eun J. K., Zhe P., Young C. Y., Young C. S. “Purification and Characterization of Chitosanase from Bacillus sp strain KCTC 0377BP and its application for the production of chitosan oligosaccharidesâ€, Applied and Environmental Microbiology, vol. 70(8), p. 4522–4531, 2004.

Sujata S., Pushplata T., and Subhash C. “A new bifunctional Chitosanase enzyme from Streptomyces sp.and its application in production of antioxidant chitooligosaccharidesâ€, Applied Biochemistry Biotechnology, vol. 167, p. 1029–1039, 2012.

Chasanah E., Gintung P., Dewi S. Z., Mahrus A., and Yenny R. “Purification and Characterization of Aeromonas media KLU 11.16 Chitosanase isolated from Shrimp Wasteâ€, Journal of Coastal Development, vol. 15(1), p. 104–113, 2011.

Lowry O., Rosebrough N., Farr A., and Randall R. “Protein measurement with the folin phenol reagentâ€, J. Biol. Chem, vol. 193, p. 265–275, 1951.

Laemmli U. K. “Cleavage of structural proteins during the assembly of the head of bacteriophage T4â€, Nature 227, p. 680–685, 1970.

Wang Y., Zhou P., Yu J.X., Pan X. R., Wang P.P., Lan W., and Tao S. “Antibacterial effect of chitooligosaccharides produced by chitosanase from Pseudomonas CUY8â€, Asia.Pac.J.Clin Nutr, vol. 16(1), p. 174–177, 2007.

Zhang H., Du Y., Yu X., Mitsutomi M., and Aiba S. “Preparation of chitooligosaccharides from chitosan by complex enzymeâ€, Carbohydr Res, vol. 320, p. 257–260, 1999.

Liu X., Xia W., Jiang Q., Xu Y., and Yu P. “Synthesis, characterization and antimicrobial activity of kojic acid grafted chitosan oligosaccharideâ€, Journal of Agricultural and Food Chem, vol. 62, p. 297–303, 2014.

Lee, D., El-Leen L., Kang M. L. “Isolation and characterization of chitosanase producing microorganism,Aureobacterium sp, Y.L. from crab shellsâ€, Journal of Microbiology and Biotechnology, vol.10(2), p. 208–214, 2000.

Shadia M., Abdel-Aziz, Hoda A. H., Foukia E. M. “Acidic Exopolysaccharide Flocculant Produced by the fungus Mucor rouxii using Beet-Molassesâ€, Research in Biotechnology, vol. 3(6), p. 01–13, 2012.

Shimosaka M., Masahiro N., Xiu- Ying W., Masanori K., chitosan –assimilating bacterium Acinetobacter sp.strain CHB101â€, Applied and Environmental Microbiology, vol. 61(2), p. 438–442, 1995.

Crispinus A. O., Naoto Y., Yoshiaki S., Kunichi K., and Kihachiro O. “Purification and some properties of a novel chitosanase from Bacillus subtilis KH1â€, J.Gen.Appl.Microbiol., vol. 46, p. 19–27, 2000.

Miller G. L. “Use of dinitrosalicylic acid reagent for determination of reducing sugarâ€, Anal. Chem. Vol. 31, p. 426–428, 1959.

Yeon J. C., Eun J. K., Zhe P., Young C. Y., and Yong C. S. “Purification and characterization of chitosanase from Bacillus sp, strain KCTC 0377BP and its application for the production of chitosan oligosaccharidesâ€, Applied and environmental Microbiology, vol. 70(8), p. 4522–4531, 2004.

Xing-Ai G, Wan-Taek J, Woo-Jin J, and Ro-Dong P. “Purification and characterization of chitosanase from Bacillus cereus D-11â€, Carbohydrate polymers, vol. 72, p. 513–520, 2008.

Chang C. L., Chang C. T., and Sung H. Y., “Purification and properties of chitosanase from a mutant of Bacillus IMR-NK1â€, Enzyme Microb Tech, vol. 32, p. 260–267, 2003.

Jo Y. Y., Jo K. J., Jin Y. I., Kim K. Y., Shim J. H., Kim Y. W., and Park R. D. “Characterization and kinetics of 45 kDa chitosanase from Bacillus sp. P16â€, Bioscience, Biotechnology and Biochemistry, vol. 67, p. 1875–1882, 2003.

Sutee W., Wanchai W., Mathana S., Nuanphan N., and Sarote S. “Production and partial characterization of chitosanases from a newly isolated Bacillus cereusâ€, Kasetsart J.(Nat.Sci), vol. 41, p. 346–355, 2007.

No H. K., Park N. Y., Lee S. H., and Meyers S. P. “Antibacterial activities of chitosan and chitosan oligomers with different molecular weightsâ€, International Journal of Food Microbiology, vol. 74, p. 65–72, 2001.

Shindia A. A., Sherbiny –EL K. A. “Chitosanase production using some Fungi optimization of fermentation conditions of chitosanase produced by Aspergillus ornatusâ€, Proceedings of the second scientific environmental conference, Zhiang University: 2007. p. 97–113.

Chuan-Lu W., Jien-Wei S., Tzu-Wen L., Anh D. N., San-Lang W. “Production, purification and characterization of a chitosanase from Bacillus cereusâ€, Journal Research on Chemical Intermediates, vol. 40(6), p. 22–37, 2014.

Kurakake M., You S., Nakagawa K., Sugihara M., and Komaki T. “Properties of chitosanase from Bacillus cereus SIâ€, Curr. Microbiol, vol. 40, p. 6–9.

You–Jin J., Pyo-Jam P., and Se-Kwon K. “Antibacterial effect of chitooligosaccharides produced by bioreactorâ€, Carbohydrate polymers, vol. 44, p. 71–76, 2001.

Xia W., Ping L., Jiali Z., and Jie C. “Biological activities of chitosan and chitooligosaccharidesâ€, Food Hydrocolloids, vol. 25, p. 170–179, 2011.

You-Jin J., Shanidi F., Pyo J. P., Se K. K. “Preparation of chitin and chitosan,chitooligosaccharides and their applications in physiological functional foodsâ€, Food International Reviews, vol. 16, p. 159–176, 2000.

Yan W., Peigen Z., Jianxing Y., Xiaorong P., Pingping W., Weiqing L., and Shendan T. “Antimicrobial effect of chitooligosaccharides produced by chitosanase from Pseudomonas CUY8â€, Asia Pac J Clin Nutr, vol. 16(1), p. 174–177, 2007.