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pubmed-article:19809241pubmed:abstractTextThe 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 protoplastregenerated 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.lld:pubmed
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pubmed-article:19809241pubmed:authorpubmed-author:KimEung-SooESlld:pubmed
pubmed-article:19809241pubmed:authorpubmed-author:LeeMi-JinMJlld:pubmed
pubmed-article:19809241pubmed:authorpubmed-author:HanKyuboemKlld:pubmed
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pubmed-article:19809241pubmed:pagination869-72lld:pubmed
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pubmed-article:19809241pubmed:year2009lld:pubmed
pubmed-article:19809241pubmed:articleTitleCombination strategy to increase cyclosporin A productivity by Tolypocladium niveum using random mutagenesis and protoplast transformation.lld:pubmed
pubmed-article:19809241pubmed:affiliationDepartment of Biological Engineering, Inha University, Incheon 402-751, Korea.lld:pubmed
pubmed-article:19809241pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:19809241pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed