Functional analysis of the validamycin biosynthetic gene cluster and engineered production of validoxylamine A.
A 45 kb DNA sequencing analysis from Streptomyces hygroscopicus 5008 involved in validamycin A (VAL-A) biosynthesis revealed 16 structural genes, 2 regulatory genes, 5 genes related transport, transposition/integration or tellurium resistance; another 4 genes had no obvious identity. The VAL-A biosynthetic pathway was proposed, with assignment of the required genetic functions confined to the sequenced region. A cluster of eight reassembled genes was found to support VAL-A synthesis in a heterologous host, S. lividans 1326. In vivo inactivation of the putative glycosyltransferase gene (valG) abolished the final attachment of glucose for VAL production and resulted in accumulation of the VAL-A precursor, validoxylamine, while the normal production of VAL-A could be restored by complementation with valG. The role of valG in the glycosylation of validoxylamine to VAL-A was demonstrated in vitro by enzymatic assay.
Bai
Linquan
L
Laboratory of Microbial Metabolism and School of Life Science & Biotechnology, Shanghai Jiaotong University, Shanghai 200030, China.
Li
Lei
L
Xu
Hui
H
Minagawa
Kazuyuki
K
Yu
Yi
Y
Zhang
Yirong
Y
Zhou
Xiufen
X
Floss
Heinz G
HG
Mahmud
Taifo
T
Deng
Zixin
Z
D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D018390:Gene Targeting; D005798:Genes, Bacterial; D005809:Genes, Regulator; D005816:Genetic Complementation Test; D005818:Genetic Engineering; D006031:Glycosylation; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism; D007294:Inositol / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D015394:Molecular Structure; D005810:Multigene Family; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism*; D048170:Streptomyces lividans / Q000235:genetics / Q000378:metabolism
D004269:DNA, Bacterial; D011994:Recombinant Proteins; C030790:validamycin A; C052534:validoxylamine A; D007294:Inositol; D016695:Glycosyltransferases