Abstract
A chitinase from Streptomyces sp. CS501 was isolated from the Korean soil sample, purified by single-step chromatography, and biochemically characterized. The extracellular chitinase (Ch501) was purified to 4.60 fold with yield of 28.74 % using Sepharose Cl-6B column. The molecular mass of Ch501 was approximately 43 kDa as estimated by SDS-PAGE and zymography. The enzyme (Ch501) was found to be stable over a broad pH range (5.0–10.0) and temperature (up to 50 °C), and have an optimum temperature of 60 °C. N-terminal sequence of Ch501 was AAYDDAAAAA. Intriguingly, Ch501 was highly sensitive to ammonium sulfate but it’s completely suppressed activity was recovered after desalting out. TLC analysis of Ch501 showed the production of N-acetyl d-glucosamine (GlcNAc) and Diacetylchitobiose (GlcNAc)2, as a principal hydrolyzed product. Ch501 shows antifungal activity against Fusarium solani and Aspergillus brasiliensis, which can be used for the biological control of fungus. As has been simple in purification, stable in a broad range of pH, ability to produce oligosaccharides, and antifungal activity showed that Ch501 has potential applications in industries as for chitooligosaccharides production used as prebiotics and/or for the biological control of plant pathogens in agriculture.
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Bhattacharya, D., A. Nagpure, and R.K. Gupta. 2007. Bacterial chitinases: Properties and potential. Critical Review in Biotechnology 27: 21–28.
Bhushan, B. 2000. Production and characterization of a thermostable chitinase from a new alkalophilic Bacillus sp. BG-11. Journal of Applied Microbiology 88: 800–808.
Bhushan, B., and G.S. Hoondal. 1998. Isolation, purification and properties of a thermostable chitinase from an alkalophilic Bacillus sp. BG-11. Biotechnology Letters 20: 157–159.
Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248–254.
Christodoulou, E., F. Duffner, and C.E. Vorgias. 2001. Overexpression, purification, and characterization of a thermostable chitinase (Chi40) from Streptomyces thermoviolaceus OPC-520. Protein Expression and Purification 23: 97–105.
Dahiya, N., R. Tewari, R.P. Tiwari, and G. Singh Hoondal. 2005. Chitinase from Enterobacter sp. NRG4: Its purification, characterization and reaction pattern. Electronic Journal of Biotechnology 8: 14–25.
Dai, D.-H., W.-L. Hu, G.-R. Huang, and W. Li. 2011. Purification and characterization of a novel extracellular chitinase from thermophilic Bacillus sp. Hu1. African Journal of Biotechnology 10: 2476–2485.
Han, Y., B. Yang, F. Zhang, X. Miao, and Z. Li. 2009. Characterization of antifungal chitinase from marine Streptomyces sp. DA11 associated with South China Sea sponge Craniella australiensis. Marine Biotechnology 11: 132–140.
Kim, K.-J., Y.-J. Yang, and J.-G. Kim. 2003. Purification and characterization of chitinase from Streptomyces sp. M-20. Journal of Biochemistry and Molecular Biology 36: 185–189.
Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.
Miller, G.L. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry 31: 426–428.
Mukherjee, G., and S.K. Sen. 2006. Purification, characterization, and antifungal activity of chitinase from Streptomyces venezuelae P10. Current Microbiology 53: 265–269.
Muzzarelli, R.A.A., J. Boudrant, D. Meyer, N. Manno, M. Demarchis, and M.G. Paoletti. 2012. Current views on fungal chitin/chitosan, human chitinases, food preservation, glucans, pectins and inulin: A tribute to Henri Braconnot, precursor of the carbohydrate polymers science, on the chitin bicentennial. Carbohydrate Polymers 87: 995–1012.
Nanjo, F., R. Katsumi, and K. Sakai. 1990. Purification and characterization of an exo-beta-d-glucosaminidase, a novel type of enzyme, from Nocardia orientalis. Journal of Biological Chemistry 265: 10088–10094.
Ober, C., and G.L. Chupp. 2009. The chitinase and chitinase-like proteins: A review of genetic and functional studies in asthma and immune-mediated diseases. Current Opinion in Allergy and Clinical Immunology 9: 401–408.
Okazaki, K., F. Kato, N. Watanabe, S. Yasuda, Y. Masui, and S. Hayakawa. 1995. Purification and properties of two chitinases from Streptomyces sp. J-13-3. Bioscience, Biotechnology, and Biochemistry 59: 1586–1587.
Ordentlich, A. 1988. The role of chitinase of Serratia marcescens in biocontrol of Sclerotium rolfsii. Phytopathology 78: 84–88.
Patil, R.S., V.V. Ghormade, and M.V. Deshpande. 2000. Chitinolytic enzymes: An exploration. Enzyme and Microbial Technology 26: 473–483.
Patil, N.S., S.R. Waghmare, and J.P. Jadhav. 2013. Purification and characterization of an extracellular antifungal chitinase from Penicillium ochrochloron MTCC 517 and its application in protoplast formation. Process Biochemistry 48: 176–183.
Pradeep, G.C., Y.H. Choi, Y.S. Choi, C.N. Seong, S.S. Cho, H.J. Lee, and J.C. Yoo. 2013a. A novel thermostable cellulase free xylanase stable in broad range of pH from Streptomyces sp. CS428. Process Biochemistry 48: 1188–1196.
Pradeep, G.C., Y.H. Choi, Y.S. Choi, S.E. Suh, J.H. Seong, S.S. Cho, M.-S. Bae, and J.C. Yoo. 2013b. An extremely alkaline novel chitinase from Streptomyces sp. CS495. Process Biochemistry 49: 223–229.
Rabeeth, M., A. Anitha, and G. Srikanth. 2011. Purification of an antifungal endochitinase from a potential biocontrol agent Streptomyces griseus. Pakistan Journal of Biological Sciences 14: 788–797.
Roberts, W.K., and C.P. Selitrennikoff. 1988. Plant and bacterial chitinases differ in antifungal activity. Journal of General Microbiology 134: 169–176.
Simkhada, J., H. Yoo, D. Park, Y. Choi, H. Lee, S. Kim, and J. Yoo. 2013. An ammonium sulfate sensitive endoxylanase produced by Streptomyces. Bioprocess and Biosystems Engineering 36: 819–825.
Tanabe, T., T. Kawase, T. Watanabe, Y. Uchida, and M. Mitsutomi. 2000. Purification and characterization of a 49 kDa chitinase from Streptomyces griseus HUT 6037. Journal of Biosciences and Bioengineering 89: 27–32.
Vieille, C., and G.J. Zeikus. 2001. Hyperthermophilic enzymes: Sources, uses, and molecular mechanisms for thermostability. Microbiology and Molecular Biology Reviews 65: 1–43.
Xiayun, J., D. Chen, H. Shenle, W. Wang, S. Chen, and S. Zou. 2012. Identification, characterization and functional analysis of a GH-18 chitinase from Streptomyces roseolus. Carbohydrate Polymers 87: 2409–2415.
Yan, Q., S. Hao, Z. Jiang, Q. Zhai, and W. Chen. 2009. Properties of a xylanase from Streptomyces matensis being suitable for xylooligosaccharides production. Journal of Molecular Catalysis B: Enzymatic 58: 72–77.
Yano, S., N. Rattanakit, A. Honda, Y. Noda, M. Wakayama, A. Plikomol, and T. Tachiki. 2008. Purification and characterization of chitinase A of Streptomyces cyaneus SP-27: An enzyme participates in protoplast formation from Schizophyllum commune mycelia. Bioscience, Biotechnology, and Biochemistry 72: 54–61.
Acknowledgments
This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korean government (MEST) (2010-0029178) and a Grant from Next-Generation BioGreen 21 Program (No. PJ009602) Rural Development Administration, Republic of Korea.
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Md. Arifur Rahman and Yun Hee Choi have contributed equally to this work.
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Rahman, M.A., Choi, Y.H., Pradeep, G.C. et al. An ammonium sulfate sensitive chitinase from Streptomyces sp. CS501. Arch. Pharm. Res. 37, 1522–1529 (2014). https://doi.org/10.1007/s12272-014-0509-z
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DOI: https://doi.org/10.1007/s12272-014-0509-z