Characterization of CalS9 in the biosynthesis of UDP-xylose and the production of xylosyl-attached hybrid compound.
The gene cluster of calicheamicin contains calS9, which encodes UDP-GlcA decarboxylase that converts UDP-GlcA to UDP-xylose. calS9 was cloned in pET32a(+) and expressed in Escherichia coli BL21 (DE3) to characterize its putative function. The reaction product was analyzed by high-performance liquid chromatography (HPLC) and electrospray ionization-mass spectrometry. The deoxysugar biosynthesis of Streptomyces sp. KCTC 0041BP was inactivated by gene replacement to generate Streptomyces sp. GerSM2 mutant, which was unable to produce dihydrochalcomycin. calS7, calS8, and calS9 UDP-xylose biosynthetic genes were cloned in an integrative plasmid pSET152 to generate pBPDS, which was heterologously expressed in Streptomyces sp. GerSM2. Finally, novel glycosylated product, 5-O-xylosyl-chalconolide derivative, in the conjugal transformants was isolated and analyzed by HPLC and liquid chromatography-mass spectrometry.
Simkhada
Dinesh
D
Institute of Biomolecule Reconstruction, Department of Pharmaceutical Engineering, SunMoon University, Kalsan-ri, Tangjeong-myeon, Asansi, Chungnam, Republic of Korea.
Oh
Tae-Jin
TJ
Pageni
Binod Babu
BB
Lee
Hei Chan
HC
Liou
Kwangkyoung
K
Sohng
Jae Kyung
JK
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002262:Carboxy-Lyases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D008969:Molecular Sequence Data; D005810:Multigene Family; D016415:Sequence Alignment; D021241:Spectrometry, Mass, Electrospray Ionization; D013302:Streptomyces / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D014540:Uridine Diphosphate Xylose / Q000096:biosynthesis*; D014994:Xylose / Q000378:metabolism*
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D014540:Uridine Diphosphate Xylose; D014994:Xylose; D002262:Carboxy-Lyases; C023257:UDPglucuronate decarboxylase