Chen Xiu-Lan, Yang Jin-Yu, Zheng Xiao-Yu, Sheng Qi, Wang Lei, Zhang Yu-Zhong, Qin Qi-Long, Zhang Xi-Ying
State Key Laboratory of Microbial Technology, Marine Biotechnology Research Center, Shandong University, Qingdao, China.
Institute of Agro-Food Science and Technology, Shandong Academy of Agricultural Sciences/Key Laboratory of Agro-Products Processing Technology of Shandong Province/Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture and Rural Affairs, Jinan, China.
Front Microbiol. 2019 Jun 26;10:1354. doi: 10.3389/fmicb.2019.01354. eCollection 2019.
Microbial extracellular proteases play crucial roles in marine protein degradation and nitrogen recycling. Although a large number of marine bacteria are found to produce extracellular proteases, it is still unknown how marine bacteria respond to environmental proteins to activate the expression of genes encoding extracellular proteases. The inducing signal molecule for marine bacterial extracellular proteases has never been identified. In this study, we identified tripeptides as the inducing signal molecules for the extracellular protease MCP-01 of the deep-sea bacterium sp. SM9913. We found that casein, but not casamino acids, can induce the gene expression and synthesis of MCP-01, suggesting that peptides rather than amino acids derived from casein induce the gene expression and synthesis of MCP-01 in SM9913. Then, casein was hydrolyzed by SM9913 extracellular proteases, and the peptides with inducing effect were isolated and characterized. Finally, four tripeptides, SPP, RYP, RQF and FRQ, were shown to have significant inducing effect on the expression of MCP-01 gene, indicating that they are likely the inducing signal molecules for the expression of protease MCP-01 gene in SM9913. This study sheds light on the induction mechanism for the gene expression and biosynthesis of marine microbial extracellular proteases, which is helpful in better understanding the adaptation of bacteria to deep-sea sedimental environment.
微生物胞外蛋白酶在海洋蛋白质降解和氮循环中发挥着关键作用。尽管发现大量海洋细菌能产生胞外蛋白酶,但海洋细菌如何响应环境中的蛋白质以激活编码胞外蛋白酶的基因表达仍不清楚。海洋细菌胞外蛋白酶的诱导信号分子从未被鉴定出来。在本研究中,我们鉴定出三肽是深海细菌sp. SM9913胞外蛋白酶MCP - 01的诱导信号分子。我们发现酪蛋白而非酪蛋白氨基酸能诱导MCP - 01的基因表达和合成,这表明源自酪蛋白的肽而非氨基酸诱导了SM9913中MCP - 01的基因表达和合成。然后,酪蛋白被SM9913胞外蛋白酶水解,分离并鉴定了具有诱导作用的肽。最后,四种三肽SPP、RYP、RQF和FRQ对MCP - 01基因的表达具有显著诱导作用,表明它们可能是SM9913中蛋白酶MCP - 01基因表达的诱导信号分子。本研究揭示了海洋微生物胞外蛋白酶基因表达和生物合成的诱导机制,有助于更好地理解细菌对深海沉积环境的适应性。