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A highly Conserved Aspartic Acid Residue of the Chitosanase from Bacillus Sp. TS Is Involved in the Substrate Binding

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Abstract

The chitosanase from Bacillus sp. TS (CsnTS) is an enzyme belonging to the glycoside hydrolase family 8. The sequence of CsnTS shares 98 % identity with the chitosanase from Bacillus sp. K17. Crystallography analysis and site-direct mutagenesis of the chitosanase from Bacillus sp. K17 identified the important residues involved in the catalytic interaction and substrate binding. However, despite progress in understanding the catalytic mechanism of the chitosanase from the family GH8, the functional roles of some residues that are highly conserved throughout this family have not been fully elucidated. This study focused on one of these residues, i.e., the aspartic acid residue at position 318. We found that apart from asparagine, mutation of Asp318 resulted in significant loss of enzyme activity. In-depth investigations showed that mutation of this residue not only impaired enzymatic activity but also affected substrate binding. Taken together, our results showed that Asp318 plays an important role in CsnTS activity.

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Acknowledgments

This work was supported, in part, by grants from the Hundred Talents Program of the Chinese Academy of Sciences (CAS), the Knowledge Innovation Program of CAS (KSCX2-EW-G-8), and the Tianjin Municipal Science and Technology Commission (10ZCKFSY05600). J.L. is an Australian National Health and Medical Council (NHMRC) Senior Research Fellow. J.S. is a recipient of the Hundred Talents Program of CAS.

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Authors and Affiliations

  1. National Engineering Laboratory for Industrial Enzymes and Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China

    Zhanping Zhou, Yang Liu, Zhengying Chang, Yanhe Ma & Jiangning Song

  2. Institute of Agro-food Science and Technology, Shandong Academy of Agricultural Sciences, Jinan, China

    Shuangzhi Zhao

  3. Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University, Vic, Parkville, 3052, Australia

    Jian Li

  4. Department of Biochemistry and Molecular Biology, Monash University, Melbourne, VIC, 3800, Australia

    Jiangning Song

  5. Monash Centre for Data Science, Faculty of Information Technology, Monash University, Melbourne, VIC, 3800, Australia

    Jiangning Song

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  1. Zhanping Zhou
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Correspondence to Jiangning Song.

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Zhou, Z., Zhao, S., Liu, Y. et al. A highly Conserved Aspartic Acid Residue of the Chitosanase from Bacillus Sp. TS Is Involved in the Substrate Binding. Appl Biochem Biotechnol 180, 1167–1179 (2016). https://doi.org/10.1007/s12010-016-2159-8

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  • DOI: https://doi.org/10.1007/s12010-016-2159-8

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