title: Mutation breeding of high 4-androstene-3,17-dione-producing Mycobacterium neoaurum ZADF-4 by atmospheric and room temperature plasma treatment.
abstract: Steroid medication is used extensively in clinical applications and comprises a large and vital part of the pharmaceutical industry. However, the difficulty of separating 4-androstene-3,17-dione (AD) from 1,4-androstadiene-3,17-dione (ADD) restricts the application of the microbial transformation of phytosterols in the industry. A novel atmospheric and room temperature plasma (ARTP) treatment, which employs helium as the working gas, was used to generate Mycobacterium neoaurum mutants producing large amounts of AD. After treatment of cultures with ARTP, four mutants were selected using a novel screening method with a color assay. Among the mutants, M. neoaurum ZADF-4 was considered the best candidate for industrial application. When the fermentation medium contained 15 g/L phytosterols and was cultivated on a rotary shaker at 160 r/min at 30 °C for 7 d, (6.28±0.11) g/L of AD and (0.82±0.05) g/L of ADD were produced by the ZADF-4 mutant, compared with (4.83±0.13) g/L of AD and (2.34±0.06) g/L of ADD by the original strain, M. neoaurum ZAD. Compared with ZAD, the molar yield of AD increased from 48.3% to 60.3% in the ZADF-4 mutant. This result indicates that ZADF-4 may have potential for industrial production of AD.
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China; Laboratory of Applied Microorganisms and Metabolic Engineering, School of Biotechnology, Jiangnan University, Wuxi 214122, China; Laboratory of Pharmaceutical Engineering, School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Liu
authors.forename: Xian
authors.initials: X
authors.lastname: Zhang
authors.forename: Zhi-ming
authors.initials: ZM
authors.lastname: Rao
authors.forename: Ming-long
authors.initials: ML
authors.lastname: Shao
authors.forename: Le-le
authors.initials: LL
authors.lastname: Zhang
authors.forename: Dan
authors.initials: D
authors.lastname: Wu
authors.forename: Zheng-hong
authors.initials: ZH
authors.lastname: Xu
authors.forename: Hui
authors.initials: H
authors.lastname: Li
mesh_terms: D000735:Androstenedione / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001272:Atmosphere; D024861:Genetic Enhancement / Q000379:methods*; D009154:Mutation / Q000235:genetics; D009161:Mycobacterium / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D058626:Plasma Gases / Q000494:pharmacology*; D013045:Species Specificity; D013696:Temperature
chemical_list: D058626:Plasma Gases; D000735:Androstenedione
keywords: 1,4-androstadiene-3,17-dione (ADD); 4-androstene-3,17-dione (AD); Atmospheric and room temperature plasma (ARTP); Mutation breeding; Mycobacterium neoaurum
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