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甘油生境解淀粉芽孢杆菌 BS-37 的基因组和转录组分析及其对 d-/l-亮氨酸的响应

Genome and transcriptome analysis of Bacillus velezensis BS-37, an efficient surfactin producer from glycerol, in response to d-/l-leucine.

机构信息

College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, China.

Oil Production Research Institute, Shengli Oil Field Ltd. Co. SinoPEC, Dongying, China.

出版信息

Microbiologyopen. 2019 Aug;8(8):e00794. doi: 10.1002/mbo3.794. Epub 2019 Feb 22.

Abstract

Surfactin is one of the most widely studied biosurfactants due to its many potential applications in different fields. In the present study, Bacillus velezensis BS-37, initially identified as a strain of Bacillus subtilis, was used to efficiently produce surfactin with the addition of glycerol, an inexpensive by-product of biodiesel production. After 36 hr of growth in glycerol medium, the total surfactin concentration reached more than 1,000 mg/L, which was two times higher than that in sucrose medium. Moreover, the addition of l- and d-Leu to the culture medium had opposite effects on surfactin production by BS-37. While surfactin production increased significantly to nearly 2,000 mg/L with the addition of 10 mM l-Leu, it was dramatically reduced to about 250 mg/L with the addition of 10 mM d-Leu. To systemically elucidate the mechanisms influencing the efficiency of this biosynthesis process, we sequenced the genome of BS-37 and analyzed changes of the transcriptome in glycerol medium in response to d-/l-leucine. The RPKM analysis of the transcriptome of BS-37 showed that the transcription levels of genes encoding modular surfactin synthase, the glycerol utilization pathway, and branched-chain amino acid (BCAA) synthesis pathways were all at a relatively high level, which may offered an explanation why this strain can efficiently use glycerol to produce surfactin with a high yield. Neither l-Leu nor d-Leu had a significant effect on the expression of genes in these pathways, indicating that l-Leu plays an important role as a precursor or substrate involved in surfactin production, while d-Leu appears to act as a competitive inhibitor. The results of the present study provide new insights into the synthesis of surfactin and ways of its regulation, and enrich the genomic and transcriptomic resources available for the construction of high-producing strains.

摘要

表面活性剂是研究最多的生物表面活性剂之一,因为它在不同领域有许多潜在的应用。在本研究中,最初被鉴定为枯草芽孢杆菌的菌株芽孢杆菌 BS-37 被用于在添加甘油(生物柴油生产的一种廉价副产物)的情况下高效生产表面活性剂。在甘油培养基中生长 36 小时后,总表面活性剂浓度超过 1000mg/L,是在蔗糖培养基中的两倍。此外,在培养基中添加 l-和 d-Leu 对 BS-37 产生表面活性剂的效果相反。虽然添加 10mM l-Leu 可使表面活性剂产量显著增加到近 2000mg/L,但添加 10mM d-Leu 会使产量急剧减少到约 250mg/L。为了系统阐明影响该生物合成过程效率的机制,我们对 BS-37 的基因组进行了测序,并分析了甘油培养基中响应 d-/l-亮氨酸时转录组的变化。BS-37 转录组的 RPKM 分析表明,编码模块化表面活性剂合成酶、甘油利用途径和支链氨基酸(BCAA)合成途径的基因的转录水平都相对较高,这可能解释了为什么该菌株可以有效地利用甘油高产表面活性剂。l-Leu 和 d-Leu 都没有对这些途径中的基因表达产生显著影响,表明 l-Leu 作为参与表面活性剂生产的前体或底物发挥重要作用,而 d-Leu 似乎作为竞争抑制剂发挥作用。本研究的结果为表面活性剂的合成及其调控方式提供了新的见解,并丰富了高产菌株构建的基因组和转录组资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8d3/6692528/85b08637ebc8/MBO3-8-e00794-g001.jpg

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