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New and highly efficient methodology for screening high‐yield strains of cytotoxic deacetylmycoepoxydiene (DAM)
Author(s) -
Zhang W.,
Wang M.,
Huang Y.,
Chea S.K.P.,
Zheng Z.,
Qian X.,
Shen Y.
Publication year - 2011
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2011.03015.x
Subject(s) - christian ministry , china , library science , chinese academy of sciences , political science , computer science , law
Aims: To establish a highly efficient methodology for screening high yield strains of cytotoxic deacetylmycoepoxydiene (DAM), to meet the need of research on its mechanism of anti‐tumor properties and in vivo toxicity studies. Methods and Results: A simple, sensitive, and highly repetitive screening procedure ‘Antimicrobial‐TLC–HPLC’ (ATH) was established for the rapid obtaining of high‐yielding DAM mutants to replace the time and labor intensive anti‐tumor activity assay (MTT). With this ATH method, four highly yielding DAM mutants were selected out of 5000 total mutants, one of which, M4‐143, showed yields of more than 300 times (250·3 mg l −1 ) that of the parent strain A123. Conclusions: The ATH method developed in this work has proven to be both economical and highly efficient with the screening of 1200 mutants in a one week time period, thusly shortening the expenditure of time and labor, without missing a single high‐yield mutant. Due to these characteristics, it is superior to other HTS screening methods described in earlier literature. The mutant M4‐143 has a good genetic stability and can be used for further research. Significance and Impact of the Study: This ATH screening method is not only perfect for screening high‐yield DAM mutants, but also, it is suitable to screen the strain libraries for those strains that have the ability to produce natural metabolites with antitumor activity.