Genetically engineered biosynthesis of macrolide derivatives including 4-amino-4,6-dideoxy-L-glucose from Streptomyces venezuelae YJ003-OTBP3.
Two sugar biosynthetic cassette plasmids were used to direct the biosynthesis of a deoxyaminosugar. The pOTBP1 plasmid containing TDP-glucose synthase (desIII), TDP-glucose-4,6-dehydratase (desIV), and glycosyltransferase (desVII/desVIII) was constructed and transformed into S. venezuelae YJ003, a strain in which the entire gene cluster of desosamine biosynthesis is deleted. The expression plasmid pOTBP3 containing 4-aminotransferase (gerB) and 3,5-epimerase (orf9) was transformed again into S. venezuelae YJ003- OTBP1 to obtain S. venezuelae YJ003-OTBP3 for the production of 4-amino-4,6-dideoxy-L-glucose derivatives. The crude extracts obtained from S. venezuelae ATCC 15439, S. venezuelae YJ003, and S. venezuelae YJ003-OTBP3 were further analyzed by TLC, bioassay, HPLC, ESI/MS, LC/MS, and MS/MS. The results of our study clearly shows that S. venezuelae YJ003-OTBP3 constructs other new hybrid macrolide derivatives including 4-amino-4,6-dideoxy-L-glycosylated YC-17 (3, [M+ Na+] m/z=464.5), methymycin (4, m/z=480.5), novamethymycin (6, m/z=496.5), and pikromycin (5, m/z=536.5) from a 12- membered ring aglycon (10-deoxymethynolide, 1) and 14-membered ring aglycon (narbonolide, 2). These results suggest a successful engineering of a deoxysugar pathway to generate novel hybrid macrolide derivatives, including deoxyaminosugar.
Pageni
Binod Babu
BB
Institute of Biomolecule Reconstruction (IBR), Department of Pharmaceutical Engineering, SunMoon University, Asan 336-708, Korea.
Oh
Tae-Jin
TJ
Liou
Kwangkyoung
K
Yoon
Yeo Joon
YJ
Sohng
Jae Kyung
JK
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D005818:Genetic Engineering / Q000379:methods*; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism; D018942:Macrolides / Q000737:chemistry / Q000378:metabolism*; D010957:Plasmids; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics*; D014170:Transformation, Genetic
C492812:4-amino-4,6-dideoxyglucose; D001426:Bacterial Proteins; D018942:Macrolides; D016695:Glycosyltransferases; D005944:Glucosamine