A High-Throughput Method for Screening of Rapamycin-Producing Strains of <Emphasis Type="Italic">Streptomyces hygroscopicus</Emphasis> by Cultivation in 96-Well Microtiter Plates
(1) Institute of Bioengineering, Department of Chemical Engineering and Bioengineering, Zhejiang University, 310027 Hangzhou, China
Abstract:
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 μg rapamycin/ml, which was double the yield of parent strain used in the submerged fermentation process.