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| Isolation, Expression and Identification of Multifunctional Chitosanase from Marine Streptomyces olivaceus FXJ7.023 |
| YUE Chang-wu1, LI Yuan-yuan1, LV Yu-hong1, WANG Miao1, SHAO Mei-yun1, LIU Ming-hao2, HUANG Ying2 |
1. Guizhou Key Laboratory of Microbial Resources & Drug Development, Zunyi Medical College, Zunyi 563003, China;
2. State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China |
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Abstract A predicted chitosanase was cloned from a fosmid genomic library of Streptomyces olivaceus strain FXJ 7.023 by PCR. The coding fragment of 885 bp encodes of 295 amino acids was fused to the expression vector pET32a (+) and transformed into Escherischia coli strains BL21 (DE3) plysS. A novel fusion protein with molecular weight of 50.3 kDa was obtained by induced the engineered strain with 1mmol/L IPTG under 18℃ and purified with the affinity chromatography of Ni2+-NTA. The recombinant chitosanase showed multifunctional catalytic activity of hydrolyze colloidal chitosan and carboxymethylcellulose with the maximal catalytic activity of 3.673U/mg and 1.302U/mg respectively. Due to it has multifunctional catalytic activity, such protein may have a potential application for the recycling of the carbohydrates and sugars in the waste.
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Received: 12 June 2014
Published: 25 August 2014
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| Fund: This work was supported by The National Basic Research Program of China (2011CB808800), The National Nature Science Foundation of China (31160004), China Ocean Mineral Resources R & D Association (DY125-15-R-02), The Science and Technology Foundation of Guizhou Province [(2010)2156] and [(2012)2348]. |
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