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Cloning and Characterization of a Chondroitin AC Exolyase from Arthrobacter sp. SD-04

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Abstract

Glycosaminoglycans (GAGs) and their low-molecular weight derivates have received considerable interest in terms of their potential clinical applications, and display a wide variety of pharmacological and pharmacokinetic properties. Structurally distinct GAG chains can be prepared by enzymatic depolymerization. A variety of bacterial chondroitin sulfate (CS) lyases have been identified, and have been widely used as catalysts in this process. Here, we identified a putative chondroitin AC exolyase gene, AschnAC, from an Arthrobacter sp. strain found in a CS manufacturing workshop. We expressed the enzyme, AsChnAC, recombinantly in Escherichia coli, then purified and characterized it in vitro. The enzyme indeed displayed exolytic cleavage activity toward HA and various CSs. Removing the putative N-terminal secretion signal peptide of AsChnAC improved its expression level in E. coli while maintaining chondroitin AC exolyase activity. This novel catalyst exhibited its optimal activity in the absence of added metal ions. AsChnAC has potential applications in preparation of low-molecular weight GAGs, making it an attractive catalyst for further investigation.

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Funding

Funding was provided by National Natural Science Foundation of China (Grant No. 31770845)

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  1. Lu-Zhou Chen and Chu-Qi Shi contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drug, School of Pharmaceutical Sciences, Shandong University, Jinan, 250012, China

    Lu-Zhou Chen, Chu-Qi Shi, Feng-Xin Yin, Feng-Shan Wang & Ju-Zheng Sheng

  2. National Glycoengineering Research Center, Shandong University, Jinan, 250012, China

    Ju-Zheng Sheng

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  1. Lu-Zhou Chen
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Chen, LZ., Shi, CQ., Yin, FX. et al. Cloning and Characterization of a Chondroitin AC Exolyase from Arthrobacter sp. SD-04. Mol Biotechnol 61, 791–800 (2019). https://doi.org/10.1007/s12033-019-00208-z

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