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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2015, Vol. 35 Issue (8): 76-82    DOI: 10.13523/j.cb.20150811
    
Screening, Identification and Enzymatic Properties Studies of Chitinase from Submarine Sediments
ZHANG Rui-xiang, YANG Zhong
School of Biotechnology, East China University of Science and Technology, Shanghai 200237, China
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Abstract  

Objective: The aim is to screen a strain with high chitinase activity from submarine sediments, to separate chitinase secreted by purified strain, and to analyze its activity by enzymatic methods. Methods: Sediments from Beibu Gulf of South China Sea were chosen as samples. Bacterial strains were screened using the clear zone and DNS method.Chitin binding assay and multiple chromatography methods were used to separate chitinase secreted by purified strain, and studied the enzymatic properties of the purified chitinase. Results: 21 strains of chitinase producers were obtained. Among them, chitinase secreted by Bacillus cereus strain B04 has the highest chitinase activity. A 36 kDa chitinase from the fermentative broth of B. cereus strain B04 was purified. Studies showed that the optimum pH and reaction temperature for the enzyme was pH 4.0 and 60℃, respectively. In pH 3.0~10.0, it has stable activity. Its activity was significantly enhanced by Co2+ and inhibited by Ag+. Conclusion:The basis for the industrial application of chitinase was provided.

Key words： Sediments      Bacillus cereus      Chitinase     
Received: 05 June 2015      Published: 25 August 2015
ZTFLH:  Q93  
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Cite this article:

ZHANG Rui-xiang, YANG Zhong . Screening, Identification and Enzymatic Properties Studies of Chitinase from Submarine Sediments. China Biotechnology, 2015, 35(8): 76-82.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20150811     OR     https://manu60.magtech.com.cn/biotech/Y2015/V35/I8/76


[1] Oh S, Kim D H, Patnaik B B, et al. Molecular and immunohistochemical characterization of the chitinase gene from Pieris rapae granulovirus. Archives of Virology, 2013, 158(8): 1701-1718.

[2] 李海浪. 壳聚糖衍生物的制备及其在药物载体中的应用研究. 上海: 中国科学院上海应用物理研究所, 2014. Li H L. Study on the preparation of chitosan derivatives and their applications in drug delivery. Shanghai: Institute of Applied Physics, Chinese Academy of Sciences, 2014.

[3] 陈海燕, 张彬. 壳寡糖的制备及其生理功能. 中国饲料, 2007, 4: 19-21. Chen H Y, Zhang B. The preparative methods and physiological function of oligochiaosan. China Feed, 2007, 4: 19-21.

[4] Bhattacharya D, Nagpure A, Gupta R K. Bacterial chitinases: properties and potential. Critical Reviews in Biotechnology, 2007, 27(1): 21-28.

[5] Rashad F M, Fathy H M, El-Zayat A S, et al. Isolation and characterization of multifunctional Streptomyces species with antimicrobial, nematicidal and phytohormone activities from marine environments in Egypt. Microbiological Research, 2015,175:34-47.

[6] Subhash Y, Sasikala C, Ramana C V. Salinimicrobium sediminis sp. nov., isolated from a deep-sea sediment. International Journal of Systematic and Evolutionary Microbiology, 2014, 64(Pt 3): 984-988.

[7] 王海东, 刘韫滔, 林伯坤, 等. 汕头海域几丁质酶产生菌的筛选及其几丁质酶编码基因的研究. 应用与环境生物学报, 2009, 15(4): 511-514. Wang H D, Liu Y T, Lin B K, et al. Isolation of chitinolytic bacteria from the Shantou Bay, China and study on their chitinase coding genes. Chinese Journal of Applied and Environmental Biology, 2009, 15(4): 511-514.

[8] 蔡亚萍, 苏建强, 谢忠, 等. 南海海域几丁质降解菌的筛选及其特性研究. 厦门大学学报(自然科学版), 2008, 47(2): 259-263. Cai Y P, Su J Q, Xie Z, et al. Isolation and characterization of chitin-degrading bacteria from South China Sea. Journal of Xiamen University(Natural Science), 2008, 47(2): 259-263.

[9] 陶树兴, 王婷婷, 徐珊, 等. 黄海沿岸及内陆盐湖几丁质酶产生菌的筛选及特性. 陕西师范大学学报(自然科学版), 2010, 38(4): 73-78. Tao S X, Wang T T, Xu S, et al. Isolation and characterization of chitinase-producing microorganisms from the Yellow Sea coast and inland salt lake. Journal of Shaanxi Normal University (Natural Science Edition), 2010, 38(4): 73-78.

[10] Kabana R R, Godoy G, Morgan J G, et al. The determination of soil chitinase activity: conditions for assay and ecological studies. Plant and Soil, 1983, 75(1): 95-106.

[11] Miller G L. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Analytical Chemistry, 1959, 31: 426-428.

[12] Kester D R, Duedall I W, Connors D N, et al. Preparation of artificial seawater. Limnology and Oceanography, 1967, 12(1): 176-179.

[13] 张军霞,丛大鹏,李雅华, 等. 棘孢木霉几丁质酶 tachi2 基因的原核表达及酶学性质研究. 中国生物工程杂志, 2013, 33(6): 45-51. Zhang J X, Cong D P, Li Y H, et al. Prokaryotic expression of tachi2 gene from Trichoderma asperellum and characterization of recombinant enzyme. China Biotechnology, 2013, 33(6): 45-51.

[14] 孙菽蔚, 王子峰, 岳海东, 等. 一种海洋几丁质酶产生菌的筛选及其产酶条件的初步研究. 海洋科学, 2007, 31(5):10-16. Sun S W, Wang Z F, Yue H D, et al. Screening of chitinase-producing marine bacterial strains and preliminary studied on chitinase-producing conditions. Marine Sciences, 2007, 31(5):10-16.

[15] Arora N, Ahmad T, Rajagopal R, et al. A constitutively expressed 36 kDa exochitinase from Bacillus thuringiensis HD-1. Biochemical and Biophysical Research Communications, 2003, 307(3): 620-625.

[16] Wang S Y, Wu S J, Thottappilly G, et al. Molecular cloning and structural analysis of the gene encoding Bacillus cereus exochitinase Chi36. Journal of Bioscience and Bioengineering, 2001, 92(1): 59-66.

[17] Hammami I, Siala R, Jridi M, et al. Partial purification and characterization of chiIO8, a novel antifungal chitinase produced by Bacillus cereus IO8. Journal of Applied Microbiology, 2013, 115(2): 358-366.

[18] Dai D H, Huang G R, Li W. Purification and characterization of a novel extracellular chitinase from thermophilic Bacillus sp. Hu1. African Journal of Biotechnology, 2011, 10(13): 2476-2485.

[19] Wang S L, Chao C H, Liang T W, et al. Purification and characterization of protease and chitinase from Bacillus cereus TKU006 and conversion of marine wastes by these enzymes. Marine Biotechnology, 2009, 11(3): 334-344.

[20] Chen W M, Chen G H, Chen C S, et al. Cloning, expression and purification of Bacillus cereus endochitinase in the Escherichia coli AD494(DE3)pLysS expression system. Bioscience Biotechnology and Biochemistry, 2009, 73(5): 1172-1174.

[21] Ferrari A R, Gaber Y, Fraaije M W. A fast, sensitive and easy colorimetric assay for chitinase and cellulase activity detection. Biotechnology for Biofuels, 2014, 7(1): 37-44.
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