P173: Molecular breeding of a fungus producing a diterpene intermediate suitable for semi-synthesis, based on the dissection of the biosynthetic machinery

Sunday, August 12, 2012
Columbia Hall, Terrace Level (Washington Hilton)
Motoyoshi Noike1, Yusuke Ono1, Nobuo Kato2 and Tohru Dairi1, (1)Graduate School of Engineering, Hokkaido University, Sapporo, Japan, (2)Institute of Scientific and Industrial Research, Osaka University, Ibaraki, Japan

Many clinically useful pharmaceuticals are semi-synthesized from natural products produced by microorganisms, which sometimes resulted in low yields and high costs. Fusicoccin A (FC) is a diterpene glycoside produced by a fungus Phomopsis amygdali, and semi-synthesized FC-derivatives without a hydroxyl group at the 12-position are now examined for application to anti-cancer drugs. We cloned FC biosynthetic genes clusters, which were scattered at two different loci, one contained four genes and another did nine genes, and characterized most of their products. Based on the results, we assumed that a mutant, in which the gene responsible for the 12-hydroxylation is disrupted, accumulates FC H, an intermediate lacking the hydroxyl group at the 12-position as a main metabolite, which can be efficiently converted into the anti-cancer drugs. The disruptant was obtained by a homologous recombination and produced FCH as a sole product as expected and its productivity was the same as that of the sum of FC-related compound produced by the parental strain. Moreover, the 19-hydroxylation gene disruptant also produced FC J as the sole compound with high titer. Taken together, the strategy, production of an intermediate compound by a gene-disruptant and its use for semi-synthesis, would be applicable for development of some pharmaceuticals, which are semi-synthesized from fungal metabolites.

(References: PNAS, 2007, 104, 3084; JACS, 2010, 133, 2548; Chembiochem, 2012, 13, 566).