Improving the production of AHL lactonase AiiO-AIO6 fromOchrobactrumsp. M231 in intracellular protease-deficientBacillus subtilis

文献类型： 外文期刊

第一作者： Xia, Rui

作者： Xia, Rui;Gao, Chenchen;Yao, Yuanyuan;Teame, Tsegay;Zhang, Fengli;Hu, Juan;Zhou, Zhigang;Yang, Yalin;Pan, Xingliang;Liu, Xuewei;Ran, Chao;Zhang, Zhen;Liu-Clarke, Jihong

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关键词： AHL lactonase; Bacillus subtilis; Quorum quenching; Intracellular protease-deletion mutant

期刊名称：AMB EXPRESS （ 影响因子：3.298； 五年影响因子：3.427 ）

ISSN： 2191-0855

年卷期： 2020 年 10 卷 1 期

页码：

收录情况： SCI

摘要： Quorum quenching (QQ) blocks bacterial cell-to-cell communication (i.e., quorum sensing), and is a promising antipathogenic strategy to control bacterial infection via inhibition of virulence factor expression and biofilm formation. QQ enzyme AiiO-AIO6 fromOchrobactrumsp. M231 has several excellent properties and shows biotherapeutic potential against important bacterial pathogens of aquatic species. AiiO-AIO6 can be secretory expressed inBacillus subtilisvia a non-classical secretion pathway. To improve AiiO-AIO6 production, four intracellular protease-deletion mutants ofB. subtilis1A751 were constructed by individually knocking out the intracellular protease-encoding genes (tepA, ymfH, yrrN and ywpE). The AiiO-AIO6 expression plasmid pWB-AIO6BS was transformed into theB. subtilis1A751 and its four intracellular protease-deletion derivatives. Results showed that all recombinant intracellular protease-deletion derivatives (BS Delta tepA, BS Delta ymfH, BS Delta yrrNand BS Delta ywpE) had a positive impact on AiiO-AIO6 production. The highest amount of AiiO-AIO6 extracellular production of BS Delta ywpEin shake flask reached 1416.47 U/mL/OD600, which was about 121% higher than that of the wild-type strain. Furthermore, LC-MS/MS analysis of the degrading products of 3-oxo-C8-HSL by purification of AiiO-AIO6 indicated that AiiO-AIO6 was an AHL-lactonase which hydrolyzes the lactone ring of AHLs. Phylogenetic analysis showed that AiiO-AIO6 was classified as a member of the alpha/beta hydrolase family with a conserved "nucleophile-acid-histidine" catalytic triad. In summary, this study showed that intracellular proteases were responsible for the reduced yields of heterologous proteins and provided an efficient strategy to enhance the extracellular production of AHL lactonase AiiO-AIO6.

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