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- Title
Development of a yeast cell based method for efficient screening of high yield tacrolimus production strain.
- Authors
Cen, Yu-Ke; He, Nai-Ying; Zhou, Wan-Ying; Liu, Zhi-Qiang; Zheng, Yu-Guo
- Abstract
Tacrolimus (FK506) is a widely used and clinically important immunosuppressant drug that can be produced by fermentation of Streptomyces tsukubaensis. The industrial strains are typically obtained through multiple rounds of mutagenesis and screening, a labor-intensive process. We have established an efficient yeast cell based screening method for the evolutionary process of high-FK506-yielding strain. The S. tsukubaensis strains of different FK506 yields were tested for zone of growth inhibition of the wild type and calcineurin mutant (cnb1∆) yeast strains. We found that different FK506 yields correspond well to altered yeast growth inhibitions. Based on the combinational inhibition effects of FK506 with different antifungals that have been frequently reported, we also tested the zone of inhibition by addition of fluconazole, amphotericin B and caspofungin to the medium. In the end, for the best screening performance, we systemically evaluated the strategy when different yeast strains and different antifungals were used according to the clarity, size, and divergence of the inhibition circles. Using different yeast strains and antifungals, we successfully broadened the screening spectrum. An efficient high-FK506-yield S. tsukubaensis screening method has been established and optimized.
- Publication
3 Biotech, 2024, Vol 14, Issue 1, p1
- ISSN
2190-572X
- Publication type
Article
- DOI
10.1007/s13205-023-03870-y