基于转录组分析揭示纳豆芽孢杆菌突变体高产甲萘醌-7的分子机理研究

闵祥兰; 赵伟; 吕小妹; 杨瑞金

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基于转录组分析揭示纳豆芽孢杆菌突变体高产甲萘醌-7的分子机理研究

首发时间：2021-04-07

闵祥兰 ¹
闵祥兰（1994-），女，硕士研究生，食品加工与配料
赵伟 ¹
赵伟(1982-), 男，博导，食品加工、蛋白质（酶）结构与功能
吕小妹 ¹ 杨瑞金 ¹

1、江南大学食品学院，无锡 214122

摘要：【目的】本文旨在通过转录组分析，揭示纳豆芽孢杆菌突变体高产甲萘醌-7（MK-7）的分子机理。【方法】首先通过UV和ARTP复合诱变，获得一株MK-7高产菌株，然后利用Illumina HiSeq 2000对诱变前后的菌株进行转录组测序，并使用Bowtie2 、RSEM、DESeq2等软件，结合GO和KEGG数据库对测序数据进行深入分析。最后，通过RT-qPCR对RNA-seq数据进行验证。【结果】获得一株MK-7产量较原始菌株提高了50.6%的突变菌株，进一步转录组分析结果表明：1）突变菌株与原始菌株相比共有1661个差异表达基因，其中上调819个，下调842个；2）在MK-7生物合成通路中甘油代谢途径和MEP途径中的一些关键基因，如glpA、dxs、ispF和ispH等显著上调；3）一些ABC转运蛋白和MFS转运蛋白基因，如msrA、bmr和bcr等以及Tat分泌途径中的基因tatA和tatC显著上调；4）大多数与孢子形成相关的基因显著下调。此外，RT-qPCR验证结果与转录组分析结果趋势一致。【结论】本研究不仅有助于我们深入了解纳豆芽孢杆菌中MK-7的合成调控机制，也为菌株的遗传改造提供了潜在的分子靶点。

关键词：纳豆芽孢杆菌甲萘醌-7 复合诱变转录组分析

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Based on transcriptome analysis reveals the molecular mechanism of high-yield menadione-7 in Bacillus subtilis natto mutant

Min Xianglan ¹
闵祥兰（1994-），女，硕士研究生，食品加工与配料
Zhao Wei ¹
赵伟(1982-), 男，博导，食品加工、蛋白质（酶）结构与功能
lyu Xiaomei ¹ Yang Ruijin ¹

1、School of Food Science and Technology, Jiangnan University，Wuxi 214122

Abstract：[Objective] The aim of this study was to reveal the molecular mechanism of the increase of menadione-7 (MK-7) production in Bacillus natto mutant by transcriptome analysis.Bacillus subtilis natto. [Methods] First, a high-yielding strain of MK-7 was obtained by UV and ARTP combined mutagenesis, and then Illumina HiSeq 2000 was used to sequence the transcriptome of the strain before and after the mutagenesis, and software such as Bowtie2, RSEM, DESeq2, etc. combined with GO and KEGG database were used to conduct in-depth analysis of the sequencing data. Finally, the RNA-seq data was verified by RT-qPCR. [Results] A mutant strain with MK-7 yield50.6%higher compared with the original strain was obtained. Further transcriptome analysis showed that: 1) Compared with the original strain, the mutant strain had a total of 1661 differentially expressed genes, of which 819 were up-regulated and 842 were down-regulated. 2) Some key genes in the glycerol metabolism pathway and MEP pathway in the MK-7 synthesis pathway, such as glpA, dxs, ispF, and ispH, are significantly up-regulated; 3) Some genes of ABC transporter and MFS transporter, such as msrA, bmr and bcr, as well as tatA and tatC in Tat secretion pathway were significantly up-regulated; 4) Most genes related to spore formation were significantly down-regulated. In addition, the RT-qPCR verification results are consistent with the trends of the transcriptome analysis results. [Conclusion] This study not only helps us to understand the synthesis mechanism of MK-7 in Bacillus subtilis natto, but also provides a potential molecular target for the genetic modification of the strain.

Keywords： Bacillus subtilis natto Menaquinone-7 Compound mutagenesis Transcriptome analysis

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闵祥兰，赵伟，吕小妹，等. 基于转录组分析揭示纳豆芽孢杆菌突变体高产甲萘醌-7的分子机理研究[EB/OL]. 北京：中国科技论文在线 [2021-04-07]. http://www.paper.edu.cn/releasepaper/content/202104-42.

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论文编号	202104-42
论文题目	基于转录组分析揭示纳豆芽孢杆菌突变体高产甲萘醌-7的分子机理研究
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