Ran Lingman, Wang Xiaolei, He Xinxin, Guo Ruihong, Wu Yanhong, Zhang Pingping, Zhang Xiao-Hua
Frontiers Science Center for Deep Ocean Multispheres and Earth System, College of Marine Life Sciences, Ocean University of China, Qingdao, China.
Laboratory for Marine Ecology and Environmental Science, Laoshan Laboratory, Qingdao, China.
Front Microbiol. 2023 Jul 3;14:1121720. doi: 10.3389/fmicb.2023.1121720. eCollection 2023.
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, 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) and minor (GlcNAc). The enzymatic activity of Chi4733 was 175.5 U mg and cat/m was 54.9 s M. Chi4733 had its maximum activity at 50°C and pH 4-6, activated by Sr, Co, Ca, and Mg and inhibited by Al, Zn, Cu, Ni, and SDS. Exo-chitinase Chi540 degraded colloidal chitin to (GlcNAc). The enzymatic activity of Chi540 was 134.5 U mg and cat/m was 54.9 s M. Chi540 had its maximum activity at 60°C and pH 6-8, was activated by Sr, Ca, and Mg but inhibited by K, Ba, Zn, Cu, Ni, SDS and urea. Whole genome analysis of WXL538 and characterization of its chitinase can provide a better understanding of its adaptability to the changing marine environment.
几丁质是海水中含量最丰富的生物聚合物,可被多种微生物用作碳源。弧菌被认为是海洋碳循环中几丁质消耗者和几丁质酶生产者的主要群体之一。这些生物广泛分布于水生环境中。然而,它们的几丁质酶之间的协同作用机制,以及几丁质酶的多样性是否有助于它们适应环境,仍需进一步阐明。在此,我们获得了一株具有八个假定几丁质酶编码基因的几丁质分解菌株WXL538。其中五个基因,即Chi4733、Chi540、Chi4668、Chi5174和Chi4963,进行了过表达和验证,其中Chi4668、Chi4733和Chi540被纯化并进行了特性分析。Chi4668的结果已在我们之前的研究中描述。内切几丁质酶Chi4733降解胶体几丁质产生(N-乙酰葡糖胺)和少量(N-乙酰葡糖胺)。Chi4733的酶活性为175.5 U mg,催化常数为54.9 s M。Chi4733在50°C和pH 4-6时具有最大活性,被Sr、Co、Ca和Mg激活,被Al、Zn、Cu、Ni和SDS抑制。外切几丁质酶Chi540将胶体几丁质降解为(N-乙酰葡糖胺)。Chi540的酶活性为134.5 U mg,催化常数为54.9 s M。Chi540在60°C和pH 6-8时具有最大活性,被Sr、Ca和Mg激活,但被K、Ba、Zn、Cu、Ni、SDS和尿素抑制。对WXL538的全基因组分析及其几丁质酶的特性分析可以更好地了解其对不断变化的海洋环境的适应性。