Function analysis of a new type I PKS-SAT domain by SAT-EAT domain replacement.
The function of a new starter unit acyltransferase (SAT) domain SAT-EF080951 (GenBank accession number) encoded in a new type I polyketide synthase (PKS) gene cluster EF568935 (GenBank accession number) isolated for this study was analyzed by domain replacement with an extender unit AT (EAT) domain of avermectin PKS. It was shown that the SAT-EF080951 incorporated malonyl-CoA specifically in vivo, which contradicted the specificity that we had previously determined by substrate binding test in vitro. The result of this study indicates that type I PKS-SAT can alter its specificity in vivo and functions well in extender units and proved the feasibility of the SAT-EAT domain replacement in type I PKS. We propose that SAT-EAT replacement strategy could be a novel route for increasing the diversity of new polyketides combinatorially biosynthesized. The new type I PKS-SAT-EF080951 studied herein may be further employed for related studies on enzymology or combinatorial biosynthesis of polyketides.
Jiao
Y L
YL
College of Marine Sciences, HuaiHai Institute of Technology, Lianyungang, China 222005. laioni1980@126.com
Wang
L H
LH
Jiao
B H
BH
Wang
S J
SJ
Fang
Y W
YW
Liu
S
S
D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020134:Catalytic Domain; D020650:Combinatorial Chemistry Techniques; D007559:Ivermectin / Q000031:analogs & derivatives / Q000378:metabolism; D018942:Macrolides / Q000378:metabolism; D054892:Metagenome; D009154:Mutation; D048630:Polyketide Synthases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013302:Streptomyces / Q000235:genetics; D014169:Transformation, Bacterial
D018942:Macrolides; D011994:Recombinant Proteins; D007559:Ivermectin; C019264:avermectin; D048630:Polyketide Synthases; D000217:Acyltransferases