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-andros...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 at- mospheric 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, Iv~. neoaururn 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.展开更多
基金supported by the National Basic Research Program(973)of China(No.2012CB725202)the National High-Tech R&D Program(863)of China(No.2011AA02A211)+4 种基金the National Natural Science Foundation of China(No.21276110)the Fundamental Research Funds for the Central Universities(Nos.JUSRP51306A and JUSRP11545)the National 111 Project of China’s Higher Education(No.111-2-06)the Program of the Key Laboratory of Industrial Biotechnology,Ministry of Education,China(No.KLIB-KF201406)the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD),China
文摘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 at- mospheric 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, Iv~. neoaururn 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.