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转录组测序揭示无色杆菌HZ01合成生物表面活性剂的潜在基因

Transcriptome Sequencing Reveals the Potential Genes of Biosurfactant Synthesis in Achromobacter sp. HZ01

  • 摘要: 为研究无色杆菌HZ01合成生物表面活性剂的分子机制,本研究将菌株HZ01分别在含柠檬酸和丙酮酸钠(对照组)的培养基中培养3 d,采用Illumina Hiseq 2500进行转录组测序。数据质控后进行序列比对、差异表达基因筛选和基因注释;利用荧光定量PCR (RT-qPCR)验证基因差异表达分析。结果显示,菌株HZ01能分别以柠檬酸和丙酮酸钠为碳源进行生长,以柠檬酸为碳源的发酵液具有良好乳化活性。表达上、下调最显著的基因分别编码OmpW和双加氧酶。“Amino acid transport and metabolism”在eggNOG数据库中富集差异表达基因的数目最多。在KEGG数据库中“,ABC transporters”富集差异表达基因的数目较多。RT-qPCR显示,基因差异表达分析的结果是可靠的。本研究鉴定出与菌株HZ01合成生物表面活性剂可能相关的基因主要包括glycosyltransferase、3-oxoacyl-ACP reductase、3-oxoacyl-ACP合成酶、硫酯酶的编码基因,推测乙醛酸循环可能在菌株HZ01的糖脂合成中发挥重要作用。本研究揭示了菌株HZ01合成生物表面活性剂的潜在基因,为后续深入研究生物表面活性剂的合成机制以及构建工程菌提供理论基础。

     

    Abstract: To study the molecular mechanisms of biosurfactant synthesis in Achromobacter sp. HZ01, strain HZ01 was cultured in the media containing citric acid and sodium pyruvate(control) for three days, respectively. The Illumina Hiseq 2500 was employed to perform transcriptome sequencing. After data quality control, sequence alignment,screening of differentially expressed genes and gene annotation were carried out. Fluorogenic quantitative PCR(RT-qPCR) was employed to verify the gene differential expression analysis. The results showed that strain HZ01 could use citric acid and sodium pyruvate as carbon source for growth, respectively. The fermentation broth using citric acid as carbon source exhibits favorable emulsifying activity. The most significantly up-regulated and down-regulated genes encoded OmpW and dioxygenase, respectively. "Amino acid transport and metabolism" contained the largest number of differentially expressed genes in the eggNOG database. “ABC transporters”contained more differentially expressed genes in the KEGG database. RT-qPCR showed that the results of gene differential expression analysis were reliable. In this study, the identified potential genes related to biosurfactant synthesis in strain HZ01 mainly included the genes coding for glycosyltransferase, 3-oxoacyl-ACP reductase, 3-oxoacyl-ACP synthetase, and thioesterase. Glyoxylate cycle may play an important role in the glycolipid synthesis in strain HZ01. This study reveals the potential genes of biosurfactant synthesis in Achromobacter sp. HZ01, laying the foundation for future in-depth studies on the molecular mechanisms of biosurfactant synthesis and construction of engineering bacteria.

     

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