用于操纵植物甾醇生物转化菌株新金色分枝杆菌NRRL B-3805的遗传技术

Genetic Techniques for Manipulation of the Phytosterol Biotransformation Strain Mycobacterium neoaurum NRRL B-3805.

作者信息

Loraine Jessica K, Smith Margaret C M

机构信息

Department of Biology, University of York, Wentworth Way, York, YO10 5DD, UK.

出版信息

Methods Mol Biol. 2017;1645:93-108. doi: 10.1007/978-1-4939-7183-1_7.

DOI:10.1007/978-1-4939-7183-1_7

PMID:28710623

Abstract

Mycobacterium neoaurum is a saprophytic, soil-dwelling bacterium. The strain NRRL B-3805 converts phytosterols to androst-4-ene-3,17-dione (androstenedione; AD), a precursor of multiple C19 steroids of importance to industry. NRRL B-3805 itself is able to convert AD to other steroid products, including testosterone (Ts) and androst-1,4-diene-3,17-dione (androstadienedione; ADD). However to improve this strain for industrial use, genetic modification is a priority. In this chapter, we describe a range of genetic techniques that can be used for M. neoaurum NRRL B-3805. Methods for transformation, expression, and gene knockouts are presented as well as plasmid maintenance and stability.

摘要

新金色分枝杆菌是一种腐生的土壤细菌。NRRL B - 3805菌株可将植物甾醇转化为雄甾-4-烯-3,17-二酮（雄烯二酮；AD），AD是多种对工业重要的C19甾体的前体。NRRL B - 3805本身能够将AD转化为其他甾体产物，包括睾酮（Ts）和雄甾-1,4-二烯-3,17-二酮（雄甾二烯二酮；ADD）。然而，为了改进该菌株以供工业使用，基因改造是首要任务。在本章中，我们描述了一系列可用于新金色分枝杆菌NRRL B - 3805的基因技术。介绍了转化、表达和基因敲除的方法以及质粒的维持和稳定性。