New and highly efficient methodology for screening high-yield strains of cytotoxic deacetylmycoepoxydiene (DAM)

作者：Zhang, W.; Wang, M.; Huang, Y.; Chea, S. K. P.; Zheng, Z.; Qian, X.^*; Shen, Y.

来源：Letters in Applied Microbiology, 2011, 52(5): 441-447.

DOI：10.1111/j.1472-765X.2011.03015.x

• cytotoxic compounds
• deacetylmycoepoxydiene (DAM)
• high-throughput screening (HTS)
• Phomopsis sp
• A123

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 center dot 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.

• 出版日期2011-5
• 单位厦门大学; 福建师范大学