Genomic analysis and chitinase characterization of Vibrio harveyi WXL538: insight into its adaptation to the marine environment

几丁质酶 甲壳素 哈维氏弧菌 微生物学 化学 生物化学 生物 海洋噬菌体 弧菌 细菌 壳聚糖 遗传学
作者
Lingman Ran,Xiaolei Wang,Xu He,Ruifeng Guo,Yanhong Wu,Pingping Zhang,Xiaohua Zhang
出处
期刊:Frontiers in Microbiology [Frontiers Media SA]
卷期号:14
标识
DOI:10.3389/fmicb.2023.1121720
摘要

Chitin, the most abundant bio-polymer in seawater, may be utilized by various microorganisms as a carbon source. Vibrios have been regarded as one of the main groups of chitin consumers in the marine carbon cycle and chitinase producers. The organisms are widely distributed in the aquatic environment. However, the co-working mechanism between their chitinases, and whether the chitinase’s diversity contributes to their adaption to the environment, needs to be further elucidated. Here, we obtained a chitinolytic strain, Vibrio harveyi WXL538 with eight putative chitinase-coding genes. Five of the genes, i.e., Chi4733, Chi540, Chi4668, Chi5174, and Chi4963, were overexpressed and validated, in which Chi4668, Chi4733 and Chi540 were purified and characterized. The result of Chi4668 was described in our previous study. Endo-chitinase Chi4733 degraded colloidal chitin to produce (GlcNAc) 2 and minor (GlcNAc) 3 . The enzymatic activity of Chi4733 was 175.5 U mg −1 and K cat/ K m was 54.9 s −1 M −1 . Chi4733 had its maximum activity at 50°C and pH 4–6, activated by Sr 2+ , Co 2+ , Ca 2+ , and Mg 2+ and inhibited by Al 3+ , Zn 2+ , Cu 2+ , Ni 2+ , and SDS. Exo-chitinase Chi540 degraded colloidal chitin to (GlcNAc) 2 . The enzymatic activity of Chi540 was 134.5 U mg −1 and K cat/ K m was 54.9 s −1 M −1 . Chi540 had its maximum activity at 60°C and pH 6–8, was activated by Sr 2+ , Ca 2+ , and Mg 2+ but inhibited by K + , Ba 2+ , Zn 2+ , Cu 2+ , Ni 2+ , SDS and urea. Whole genome analysis of V. harveyi WXL538 and characterization of its chitinase can provide a better understanding of its adaptability to the changing marine environment.
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