Journal
BIOTECHNOLOGY LETTERS
Volume 27, Issue 15, Pages 1135-1140Publisher
SPRINGER
DOI: 10.1007/s10529-005-8463-y
Keywords
bioassay; high-throughput method; rapamycin; Streptomyces hygroscopicus; 96-well microtiter plate
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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 mu g rapamycin/ml, which was double the yield of parent strain used in the submerged fermentation process.
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