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Molecular characterization of an endo-type chitosanase from the fish pathogen Renibacterium sp. QD1

Published online by Cambridge University Press:  28 January 2014

Peichuan Xing ,
Dan Liu ,
Wen-Gong Yu  and
Xinzhi Lu
Peichuan Xing
Affiliation:
Laboratory of Marine Drugs of Chinese Ministry of Education, Ocean University of China, 5 Yushan Road, Qingdao 266003, China Shandong Provincial Key Laboratory of Glycoscience & Glycotechnology, Ocean University of China, 5 Yushan Road, Qingdao 266003, China School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, China
Dan Liu
Affiliation:
Shandong Provincial Key Laboratory of Glycoscience & Glycotechnology, Ocean University of China, 5 Yushan Road, Qingdao 266003, China School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, China
Wen-Gong Yu
Affiliation:
Shandong Provincial Key Laboratory of Glycoscience & Glycotechnology, Ocean University of China, 5 Yushan Road, Qingdao 266003, China School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, China
Xinzhi Lu*
Affiliation:
Shandong Provincial Key Laboratory of Glycoscience & Glycotechnology, Ocean University of China, 5 Yushan Road, Qingdao 266003, China School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, China
*
Correspondence should be addressed to: X. Lu, Department of Biology Technology, School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, China. email: lxinzhi@gmail.com
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Abstract

Renibacterium sp. QD1, a bacteria strain capable of hydrolysing chitosan, was isolated from the homogenate of small crabs. An extracellular chitosanase, Csn-A, was purified from the QD1 fermentation broth. The enzyme was purified to homogeneity, with a yield of eight-fold, 67% recovery and a specific activity of 1575 U/mg proteins. The molecular weight of Csn-A was estimated to be 26.1 kDa by SDS-PAGE. Unlike other chitosanases, the purified Csn-A displayed maximal activity at a pH range of 5.3–6.5, and it was stable in a broad pH range of 5.0–10.0. The optimum temperature for chitosanlytic activity was 55°C. The enzyme activity was strongly stimulated by Mn2+ but inhibited by Fe3+, Cu2+, Al3+, Zn2+ and SDS. TLC analysis demonstrated that Csn-A hydrolysed N-deacetylated polymeric glucosamines into chito-biose and -triose in an endo-type manner. The amino acid seuquence of Csn-A showed close identity with an uncharacterized chitosanase of strain ATCC33209.

Keywords

characterizationChitosanaseChitooligosaccharidesRenibacterium sp. QD1
Type
Research Article
Information
Journal of the Marine Biological Association of the United Kingdom , Volume 94 , Issue 4 , June 2014 , pp. 681 - 686
Copyright
Copyright © Marine Biological Association of the United Kingdom 2014 

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