earticle

영어

The cyclic undecapeptide, cyclosporin A (CyA), one of the most valuable immunosuppressive drugs, is produced non-ribosomally by a multi-functional cyclosporin synthetase enzyme complex by the filamentous fungus Tolypocladium niveum. The various structural modifications of cyclosporins such as a regiospecific hydroxylation at N-methyl leucines lead to a dramatic changes of their bioactive spectra. A Rare Actinomycetes named Sebekia benihana was identified to be able to hydroxylate CyA at the position of 4th Nmethyl leucine. Previously, six different novel Cytochrome P450 hydroxylase (CYP) genes have been isolated from S. benihana, followed by the complete sequencing and characterization1). Since a rare actinomycetes speices including, S. benihana is not currently feasible for genetic manipulation, we tried to establish genome engineering techniques for S. benihana in order to maximize its potential as a bioconversion host of various valuable metabolites including CyA in vivo. Detailed results including interspecies conjugation, foreign gene-integration, and a targeted-gene-disruption in S. benihana will be discussed.