拟无枝酸菌(Amycolatopsis sp.)CCTCC NO:M2011265是一株可以转化阿魏酸生成香兰素菌株,该研究利用Illumina Hiseq测序平台对拟无枝酸菌进行全基因组测序、拼接、基因预测及功能注释,并从拟无枝酸菌的全基因组中筛选和鉴定参与香兰素合成的功能基因。结果表明,总共组装得到64个scaffolds,整个基因组组装大小约为8 425 551 bp,总GC含量为71.89%。通过生物信息学分析发现了香兰素合成关键基因ech、fcs和ech2基因,及香兰素分解代谢基因vdh基因,其中ech和fcs为一个基因簇,并进一步构建过表达ech-fcs-ech2菌株,其摇瓶发酵表明转化阿魏酸生成香兰素速率加快,发酵时间显著缩短。该研究获得的拟无枝酸菌的全基因组信息为解析其转化阿魏酸生成香兰素发酵过程中的代谢机理提供遗传信息基础,也为通过代谢工程获得高产香兰素菌株的研究提供理论支持。
Amycolatopsis sp. CCTCC NO: M2011265 is a strain that can biotransform ferulic acid to produce vanillin. In this study, the Illumina HiSeq sequencing platform was used to perform whole genome sequencing,splicing, gene prediction, and functional annotation were performed to screen and identify functional genes involved in vanillin biosynthesis from the whole genome of Amycolatopsis sp. The results showed that a total of 64 scaffolds were assembled, with the whole genome assembly size of about 8 425 551 bp and the total GC content of 71.89%. Through bioinformatics analysis, the key genes of vanillin synthesis including ech, fcs, and ech2, and the vanillin degradation gene vdh gene were found, in which ech-fcs belonged to a gene cluster. In addition, the overexpression strain of ech-fcs-ech2 was constructed, and the shaker fermentation showed that the conversion of ferulic acid to vanillin was accelerated and the fermentation time was significantly shortened. The whole genome information obtained in this study provides a basis of genetic information for understanding the metabolic mechanism of Amycolatopsis sp. during the fermentative production vanillin from ferulic acid, and also provides theoretical support for the research of obtaining high vanillin-producing strains through metabolic engineering.
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