ISOLATION, SCREENING, AND CHARACTERIZATION OF CHITINASE PRODUCING BACTERIA FROM MARINE WASTES

Authors

  • S. Krithika
  • C. Chellaram Vel Tech Multitech Engineering College, Chennai-600062, Tamil Nadu, India, Applied Biotechnology Department, Sur College of Applied Sciences, Sur-411, Oman

Abstract

Objective: Aim of this study deals with screening and characterization of chitinase-producing bacteria from marine waste and its deposited soil along the coastal regions in Chennai.

Methods: The soil samples were collected aseptically and subjected to serial dilution to isolate the bacterial strains. Totally, 35 morphologically different microorganisms were isolated and were screened for their chitinolytic activity in colloidal chitin incorporated media through zone assay using Congo red stain. The biochemical tests were performed for the isolated to prove their validity and further with sequencing to determine the species.

Results: The isolates were screened based on the size of the zone formed. Best chitinase producers were subjected to biochemical tests and 16s ribosomal RNA sequencing. A novel strain, Acinetobacter ASK18, a gram-negative, motile organism was identified. Thus, the isolate may be a potent producer of chitinase, and the marine wastes can be utilized efficiently to generate a high value-added product.

Conclusion: A novel strain, Acinetobacter ASK18, would further be subjected to purification of the enzyme produced, and hence the active principle could be evaluated as an effective pharmacological drug in anticancer and antibacterial properties.

Keywords: Marine wastes, Chitinase, Congo red, Biochemical tests, 16s rRNA sequencing

Downloads

Download data is not yet available.

References

Patil RS, Ghormade VV, Deshpande MV. Chitinolytic enzymes: an exploration. Enzyme Microb Technol 2000;26 Suppl 7:473-83.

Svitil AL, Chadhain S, Moore JA, Kirchman DL. Chitin degradation proteins produced by the marine bacterium Vibrio harveyi growing on different forms of chitin. Appl Environ Microbiol 1997;63 Suppl 2:408-13.

Wang SL, Lin HT, Liang TW, Chen YJ, Yen YH, Guo SP. Reclamation of chitinous materials by bromelain for the preparation of antitumor and antifungal materials. Bioresour Technol 2008;99 Suppl 10:4386-93.

Shanmugaiah V, Mathivanan N, Balasubramanian N, Manoharan P. Optimization of cultural conditions for production of chitinase by Bacillus laterosporous isolated from rice rhizosphere soil. Afr J Biotechnol 2008;7:2562-8.

Ajit NS, Verma R, Shanmugam V. Extracellular chitinases of fluorescent pseudomonads antifungal to Fusarium oxysporum f. sp. dianthi causing carnation wilt. Curr Microbiol 2006;52 Suppl 4:310-6.

Akagi K, Watanabe J, Hara M, Kezuka Y, Chikaishi E, Yamaguchi T, et al. Identification of the substrate interaction region of the chitin-binding domain of Streptomyces griseous chitinase C. J Biochem 2006;139 Suppl 3:483-93.

Viterbo A, Haran S, Friesem D, Ramot O, Chet I. Antifungal activity of a novel endochitinase gene (chit36) from Trichoderma harzianum Rifai TM. FEMS Microbiol Lett 2001;200 Suppl 2:169-74.

Matsushima R, Ozawa R, Uefune M, Gotoh T, Takabayashi J. Intraspecies variation in the kanzawa spider mite differentially affects induced defensive response in lima bean plants. J Chem Ecol 2006;32 Suppl 11:2501-12.

Vyas PR, Deshpande MV. Enzymatic hydrolysis of chitin by Myrothecium verrucaria chitinase complex and its utilization to produce SCP. J Gen Appl Microbiol 1991;37:267.

Ferrer J, Paez G, Marmol Z, Ramones E, Garcia H, Forster CF. Acid hydrolysis of shrimp shell waste and the production of single cell protein from the hydrolysate. Bioresour Technol 1996;57 Suppl 1:55-60.

Felse PA, Panda T. Production of microbial chitinases-a revisit. Bioprocess Eng 2000;23 Suppl 2:127-34.

Prabavathy VR, Mathivanan N, Sagadevan E, Murugesan K, Lalithakumari D. Intra-strain protoplast fusion enhances carboxymethyl cellulase activity in Trichoderma reesei. Enzyme Microb Technol 2006;38 Suppl 5:719-23.

Chang WT, Chen YC, Jao CL. Antifungal activity and enhancement of plant growth by Bacillus cereus grown on shellfish chitin wastes. Bioresour Technol 2007;98 Suppl 6:1224-30.

Chakrabortty S, Bhattacharya S, Das A. Optimization of process parameters for chitinase production by a marine isolate of Serratia marcescens. Int J Pharma Bio Sci 2012;2:8-20.

Skujins JJ, Pogieter HJ, Alexander M. Dissolution of fungal walls by a Streptomycete-chitinase and glucannase. Arch Biochem Biophys 1965;111:358-64.

Prem Anand, Chellaram C, Shanthini F. Isolation, and screening of marine bacteria producing antibiotics against human pathogens. Int J Pharma Biosci Technol 2011;2:1-15.

Keddie RM, Shaw S, Kurthia G. In: Bergey’s Manual of Systematic Bacteriology. Sneath PH, Mair N, Sharpe ME, Holt JG. Eds. Baltimore: Williams and Wilkins; 1986;2:1255-8.

Sambrook J, Russell DW. Molecular cloning: a laboratory manual. Vol. 1. 3rd ed. Cold Spring Harbor. NY: Cold Spring Harbor Laboratory; 2001;4:53,8:30-34.

Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhan Z, Miller W, et al. Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucleic Acids Res 1997;25 Suppl 17:3389-402.

Frändberg E. Antifungal compounds of chitinolytic bacteria from grain ecosystems. Doctor’s Dissertation; 1997.

Henrissat B. Classification of glycosyl hydrolases are based on amino acid sequence similarities. Biochem J 1991;280:309-16.

Henrissat B, Bairoch A. New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J 1993;293:781-8.

Watanabe T, Kobori K, Miyashita K, Fujii T, Sakai H, Uchida M, et al. Identification of glutamic acid 204 and aspartic acid 200 in chitinase A1 of Bacillus circulans WL-12 as essential residues for chitinase activity. J Biol Chem 1993;268:18567-72.

Chigaleichik AG, Pirieva DA, Rylkin SS. Chitinases from Serratia marcescens Prikladnaya. Biochim Mikrobiol 1976;12:581-6.

Krishnan HB, Kim KY, Krishnan AH. Expression of a Serratia marcescens chitinase gene in Sinorhizobium fredii USDA191 and Sinorhizobium meliloti RCR2011 impedes soybean and alfalfa nodulation. Mol Plant Microbe Interact 1999;12 Suppl 8:748–51.

Das MP, Rebecca LJ, Sharmila S, Anu, Banerjee A, Kumar D. Identification and optimization of cultural conditions for chitinase production by Bacillus amyloliquefaciens SM3. J Chem Pharm Res 2012;4 Suppl 11:4816-21.

Published

01-05-2016

How to Cite

Krithika, S., and C. Chellaram. “ISOLATION, SCREENING, AND CHARACTERIZATION OF CHITINASE PRODUCING BACTERIA FROM MARINE WASTES”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 8, no. 5, May 2016, pp. 34-36, https://journals.innovareacademics.in/index.php/ijpps/article/view/10109.

Issue

Section

Original Article(s)