Characterization of the polyoxin biosynthetic gene cluster from Streptomyces cacaoi and engineered production of polyoxin H.
A gene cluster (pol) essential for the biosynthesis of polyoxin, a nucleoside antibiotic widely used for the control of phytopathogenic fungi, was cloned from Streptomyces cacaoi. A 46,066-bp region was sequenced, and 20 of 39 of the putative open reading frames were defined as necessary for polyoxin biosynthesis as evidenced by its production in a heterologous host, Streptomyces lividans TK24. The role of PolO and PolA in polyoxin synthesis was demonstrated by in vivo experiments, and their functions were unambiguously characterized as O-carbamoyltransferase and UMP-enolpyruvyltransferase, respectively, by in vitro experiments, which enabled the production of a modified compound differing slightly from that proposed earlier. These studies should provide a solid foundation for the elucidation of the molecular mechanisms for polyoxin biosynthesis, and set the stage for combinatorial biosynthesis using genes encoding different pathways for nucleoside antibiotics.
Chen
Wenqing
W
Laboratory of Microbial Metabolism and School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai 200030, China.
Huang
Tingting
T
He
Xinyi
X
Meng
Qingqing
Q
You
Delin
D
Bai
Linquan
L
Li
Jialiang
J
Wu
Mingxuan
M
Li
Rui
R
Xie
Zhoujie
Z
Zhou
Huchen
H
Zhou
Xiufen
X
Tan
Huarong
H
Deng
Zixin
Z
D002874:Chromosome Mapping; D005816:Genetic Complementation Test; D005818:Genetic Engineering / Q000379:methods*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D005810:Multigene Family; D016296:Mutagenesis; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D011741:Pyrimidine Nucleosides / Q000096:biosynthesis / Q000235:genetics; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism
D020033:Protein Isoforms; D011741:Pyrimidine Nucleosides; C002457:polyoxin