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新金色分枝杆菌中用于生产有价值甾体中间体的单核苷酸多态性分析

Single nucleotide polymorphism analysis for the production of valuable steroid intermediates in Mycobacterium neoaurum.

作者信息

Liu Min, Zhu Zhan-Tao, Tao Xin-Yi, Wang Feng-Qing, Wei Dong-Zhi

机构信息

State Key Lab of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China.

出版信息

Biotechnol Lett. 2016 Nov;38(11):1881-1892. doi: 10.1007/s10529-016-2187-z. Epub 2016 Aug 29.

Abstract

OBJECTIVES

To investigate single nucleotide polymorphism (SNP) in the transformation process of phytosterol to valuable steroid intermediates in three steroid-producing Mycobacterium neoaurum strains using deep sequencing and bioinformation analysis.

RESULTS

The assembled contig sequences from RNA sequencing of strains producing 9α-hydroxy-4-androstene-3,17-dione (9OHAD), 1,4-androstadiene-3,17-dione (ADD), and 22-hydroxy-23, 24-bisnorchola-1,4-dien-3-one (1,4-BNA) were analyzed for the presence of putative SNPs for steroid catabolism. 413, 375, and 491 SNPs were detected in the coding domain sequences and non-coding domain sequences of RNA sequencing reads of M. neoaurum strains producing 9OHAD, ADD, and BNA, respectively. Special attention was focused on SNPs associated with genes showing differential expression at proteome level, including the genes for sterol catabolism, glycerol catabolic process, signal transduction systems, transport system and energy metabolism.

CONCLUSIONS

The work facilitates the understanding of underlying genetic changes that may be responsible for steroid accumulation in M. neoaurum and is useful for its targeted genetic engineering.

摘要

目的

利用深度测序和生物信息分析,研究三种产甾体的新金色分枝杆菌菌株中植物甾醇转化为有价值的甾体中间体过程中的单核苷酸多态性(SNP)。

结果

对产生9α-羟基-4-雄甾烯-3,17-二酮(9OHAD)、1,4-雄甾二烯-3,17-二酮(ADD)和22-羟基-23,24-双降胆甾-1,4-二烯-3-酮(1,4-BNA)的菌株的RNA测序组装的重叠群序列进行分析,以寻找甾体分解代谢的推定SNP。在产生9OHAD、ADD和BNA的新金色分枝杆菌菌株的RNA测序读数的编码域序列和非编码域序列中分别检测到413、375和491个SNP。特别关注与在蛋白质组水平上显示差异表达的基因相关的SNP,包括甾醇分解代谢、甘油分解代谢过程、信号转导系统、转运系统和能量代谢的基因。

结论

这项工作有助于理解可能导致新金色分枝杆菌中甾体积累的潜在遗传变化,对其靶向基因工程有用。

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