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

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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.

Original languageEnglish
Title of host publicationMicrobial Steroids
EditorsJosé-Luis Barredo, Ignacio Herráiz
Number of pages16
ISBN (Electronic)978-1-4939-7183-1
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Journal Article

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