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Characterization of Antifungal Chitinase from Marine Streptomyces sp. DA11 Associated with South China Sea Sponge Craniella Australiensis

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Abstract

The gene cloning, purification, properties, kinetics, and antifungal activity of chitinase from marine Streptomyces sp. DA11 associated with South China sponge Craniella australiensis were investigated. Alignment analysis of the amino acid sequence deduced from the cloned conserved 451 bp DNA sequence shows the chitinase belongs to ChiC type with 80% similarity to chitinase C precursor from Streptomyces peucetius. Through purification by 80% ammonium sulfate, affinity binding to chitin and diethylaminoethyl-cellulose anion-exchange chromatography, 6.15-fold total purification with a specific activity of 2.95 Umg−1 was achieved. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed a molecular weight of approximately 34 kDa and antifungal activities were observed against Aspergillus niger and Candida albicans. The optimal pH, temperature, and salinity for chitinase activity were 8.0, 50°C, and 45 g‰ psu, respectively, which may contribute to special application of this marine microbe-derived chitinase compared with terrestrial chitinases. The chitinase activity was increased by Mn2+, Cu2+, and Mg2+, while strongly inhibited by Fe2+ and Ba2+. Meanwhile, SDS, ethyleneglycoltetraacetic acid, urea, and ethylenediaminetetraacetic acid were found to have significantly inhibitory effect on chitinase activity. With colloidal chitin as substrates instead of powder chitin, higher V max (0.82 mg product/min·mg protein) and lower K m (0.019 mg/ml) values were achieved. The sponge’s microbial symbiont with chitinase activity may contribute to chitin degradation and antifungal defense. To our knowledge, it was the first time to study sponge-associated microbial chitinase.

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Acknowledgements

Financial support to Zhiyong Li from “High-Tech Research and Development Program of China” (2007AA09Z447) is greatly acknowledged. Shanghai Leading Academic Discipline Project (B203) provides the related research platform. Thanks for Professor Rongxiu Li for his help in the enzyme purification and SDS-PAGE analysis.

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  1. Laboratory of Marine Biotechnology, School of Life Science and Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, 200240, People’s Republic of China

    Yue Han, Bingjie Yang, Fengli Zhang, Xiaoling Miao & Zhiyong Li

  2. Key Laboratory of Microbial Metabolism, Ministry of Education, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai, 200240, People’s Republic of China

    Fengli Zhang, Xiaoling Miao & Zhiyong Li

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  1. Yue Han
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  2. Bingjie Yang
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Correspondence to Zhiyong Li.

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Han, Y., Yang, B., Zhang, F. et al. Characterization of Antifungal Chitinase from Marine Streptomyces sp. DA11 Associated with South China Sea Sponge Craniella Australiensis . Mar Biotechnol 11, 132–140 (2009). https://doi.org/10.1007/s10126-008-9126-5

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