A high-throughput method for screening of rapamycin-producing strains of Streptomyces hygroscopicus by cultivation in 96-well microtiter plates.
A novel high-throughput cultivation method was developed to rapidly screen large numbers of rapamycin-producing mutants of Streptomyces hygroscopicus by duplicate culturing of isolates on the surfaces of agar-solidified 96 wells in microtiter plates. One copy of the cultures was used for the rapamycin bioassay and the other identical copy, representing potentially high yielding strains, was preserved for further study. By integrating 96-well solid cultivation and the bioassay, we screened more than 7000 isolates and found 10 high-yielding strains. From these, one mutant produced 420 mug rapamycin/ml, which was double the yield of parent strain used in the submerged fermentation process.
Xu
Zhi-nan
ZN
Institute of Bioengineering, Department of Chemical Engineering and Bioengineering, Zhejiang University, 310027, Hangzhou, China. znxu@zju.edu.cn
Shen
Wen-he
WH
Chen
Xi-yang
XY
Lin
Jian-ping
JP
Cen
Pei-lin
PL
D000362:Agar / Q000737:chemistry; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001681:Biological Assay; D001709:Biotechnology / Q000379:methods*; D002176:Candida albicans / Q000378:metabolism; D005285:Fermentation; D009154:Mutation; D020123:Sirolimus / Q000494:pharmacology*; D013302:Streptomyces / Q000378:metabolism*; D013997:Time Factors
D000900:Anti-Bacterial Agents; D000362:Agar; D020123:Sirolimus