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Combination Strategy to Increase Cyclosporin A Productivity by Tolypocladium niveum Using Random Mutagenesis and Protoplast Transformation  

Lee, Mi-Jin (Department of Biological Engineering, Inha University)
Duong, Cae Thi Phung (Department of Biological Engineering, Inha University)
Han, Kyu-Boem (Hanson Biotech Co., Ltd)
Kim, Eung-Soo (Department of Biological Engineering, Inha University)
Publication Information
Journal of Microbiology and Biotechnology / v.19, no.9, 2009 , pp. 869-872 More about this Journal
Abstract
The cyclic undecapeptide cyclosporin A (CyA), one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex by the filamentous fungus Tolypocladium niveum. To increase CyA productivity by wild-type T. niveum (ATCC 34921), random mutagenesis was first performed using an antifungal agar-plug colony assay (APCA) selection approach. This generated a mutant strain producing more than 9-fold greater CyA than the wild-type strain. Additionally, a foreign bacterial gene, Vitreoscilla hemoglobin gene (VHb), was transformed via protoplast regeneration and its transcription was confirmed by RT-PCR in the UV-irradiated mutant cell. This led to an additional 33.5% increase of CyA production. Although most protoplast-regenerated T. niveum transformants tend to lose CyA productivity, the optimized combination of random mutagenesis and protoplast transformation described here should be an efficient strategy to generate a commercially valuable, yet metabolite low-producing, fungal species, such as CyA-producing T. niveum.
Keywords
Tolypocladium niveum; cyclosporin A; mutagenesis; protoplast transformation;
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Times Cited By KSCI : 2  (Citation Analysis)
Times Cited By Web Of Science : 3  (Related Records In Web of Science)
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