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一种来自深海细菌QD07的新型壳聚糖酶的克隆与特性分析

Cloning and Characterization of a New Chitosanase From a Deep-Sea Bacterium sp. QD07.

作者信息

Zheng Qiuling, Meng Xiangjun, Cheng Mingyang, Li Yanfeng, Liu Yuanpeng, Chen Xuehong

机构信息

Department of Pharmacology, School of Basic Medicine, Qingdao University, Qingdao, China.

Qingdao Mental Health Center, Qingdao, China.

出版信息

Front Microbiol. 2021 Feb 24;12:619731. doi: 10.3389/fmicb.2021.619731. eCollection 2021.

DOI:10.3389/fmicb.2021.619731
PMID:33717008
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7943732/
Abstract

Chitosanase is a significant chitosan-degrading enzyme involved in industrial applications, which forms chitooligosaccharides (COS) as reaction products that are known to have various biological activities. In this study, the gene was cloned from a deep-sea bacterium sp. QD07, as well as over-expressed in , which is a new chitosanase encoding gene. The recombinant strain was cultured in a 5 L fermenter, which yielded 324 U/mL chitosanases. After purification, CsnS is a cold-adapted enzyme with the highest activity at 60°C, showing 37.5% of the maximal activity at 0°C and 42.6% of the maximal activity at 10°C. It exhibited optimum activity at pH 5.8 and was stable at a pH range of 3.4-8.8. Additionally, CsnS exhibited an endo-type cleavage pattern and hydrolyzed chitosan polymers to yield disaccharides and trisaccharides as the primary reaction products. These results make CsnS a potential candidate for the industrial manufacture of COS.

摘要

壳聚糖酶是一种参与工业应用的重要壳聚糖降解酶，它形成的反应产物壳寡糖（COS）具有多种生物活性。在本研究中，该基因从深海细菌QD07中克隆出来，并在大肠杆菌中过表达，这是一个新的壳聚糖酶编码基因。重组菌株在5 L发酵罐中培养，产壳聚糖酶的量为324 U/mL。纯化后，CsnS是一种冷适应酶，在60°C时活性最高，在0°C时显示出最大活性的37.5%，在10°C时显示出最大活性的42.6%。它在pH 5.8时表现出最佳活性，在pH 3.4 - 8.8范围内稳定。此外，CsnS表现出内切型切割模式，水解壳聚糖聚合物产生二糖和三糖作为主要反应产物。这些结果使CsnS成为工业生产壳寡糖的潜在候选者。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/cd8a9a8e16cf/fmicb-12-619731-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/4516ae2e4335/fmicb-12-619731-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/26b142d17c6d/fmicb-12-619731-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/cfd4601a4081/fmicb-12-619731-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/f8c952b23dd2/fmicb-12-619731-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/7abb4c9c7465/fmicb-12-619731-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/cd8a9a8e16cf/fmicb-12-619731-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/4516ae2e4335/fmicb-12-619731-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/26b142d17c6d/fmicb-12-619731-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/cfd4601a4081/fmicb-12-619731-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/f8c952b23dd2/fmicb-12-619731-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/7abb4c9c7465/fmicb-12-619731-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f55/7943732/cd8a9a8e16cf/fmicb-12-619731-g006.jpg

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