Application of 2,4-Dinitrophenylhydrazine (DNPH) in High-Throughput Screening for Microorganism Mutants Accumulating 9α-Hydroxyandrost-4-ene-3,17-dione (9α-OH-AD).
作者： Yang Liu , Fei Cao , Hui Xiong , Yanbing Shen , Min Wang
DOI: 10.1371/JOURNAL.PONE.0163836
关键词:
摘要: To develop a quick method for the preliminarily screening of mutant strains that can accumulate 9α-hydroxyandrost-4-ene-3,17-dione (9α-OH-AD), high-throughput was presented by applying principle 2,4-dinitrophenylhydrazine (DNPH) react with ketones to produce precipitation. The optimal color assay conditions were substrate androst-4-ene-3,17-dione (AD) concentration at 2.0 g/L, ratio AD DNPH solution 1:4, and sulfuric acid ethanol percentages in 2% 35%, respectively. This used screen mutants Rhodococcus rhodochrous DSM43269, from which three ones obtained could more 9α-OH-AD. not only broadens methods increases efficiency microbial mutation breeding but also establishes good foundation obtaining industrial application.
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