Cloning and characterization of monacolin K biosynthetic gene cluster from Monascus pilosus.
Monacolin K is a secondary metabolite synthesized by polyketide synthases (PKS) from Monascus, and it has the same structure as lovastatin, which is mainly produced by Aspergillus terreus. In the present study, a bacterial artificial chromosome (BAC) clone, mps01, was screened from the BAC library constructed from Monascus pilosus BCRC38072 genomic DNA. The putative monacolin K biosynthetic gene cluster was found within a 42 kb region in the mps01 clone. The deduced amino acid sequences encoded by the nine genes designated as mokA- mokI, which share over 54% similarity with the lovastatin biosynthetic gene cluster in A. terreus, were assumed to be involved in monacolin K biosynthesis. A gene disruption construct designed to replace the central part of mokA, a polyketide synthase gene, in wild-type M. pilosus BCRC38072 with a hygromycin B resistance gene through homologous recombination, resulted in a mokA-disrupted strain. The disruptant did not produce monacolin K, indicating that mokA encoded the PKS responsible for monacolin K biosynthesis in M. pilosus BCRC38072.
Chen
Yi-Pei
YP
Bioresource Collection and Research Center, Food Industry Research and Development Institute, National Chiao Tung University, HsinChu, Taiwan.
Tseng
Ching-Ping
CP
Liaw
Li-Ling
LL
Wang
Chun-Lin
CL
Chen
I-Ching
IC
Wu
Wen-Jung
WJ
Wu
Ming-Der
MD
Yuan
Gwo-Fang
GF
D000595:Amino Acid Sequence; D022202:Chromosomes, Artificial, Bacterial; D003001:Cloning, Molecular; D008148:Lovastatin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D032741:Monascus / Q000201:enzymology / Q000235:genetics*; D010802:Phylogeny; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism
D048630:Polyketide Synthases; D008148:Lovastatin