Enzymatic synthesis of dTDP-4-amino-4,6-dideoxy-D-glucose using GerB (dTDP-4-keto-6-deoxy-D-glucose aminotransferase).
Over-expressed GerB (dTDP-4-keto-6-deoxy-d-glucose aminotransferase) of Streptomyces sp. GERI-155 was used in the enzymatic synthesis of dTDP-4-amino-4,6-dideoxy-D-glucose (2) from dTDP-4-keto-6-deoxy-D-glucose (1). [Carbohydrate structure: see text]. Five enzymes including dTMP kinase (TMK), acetate kinase (ACK), dTDP-glucose synthase (TGS), dTDP-glucose 4,6-dehydratase (DH), and dTDP-4-keto-6-deoxy-d-glucose aminotransferase (GerB) were used to synthesize 2 on a large scale from glucose-1-phosphate and TMP. A conversion yield of up to 57% was obtained by HPLC peak integration given a reaction time of 270min. After purification by two successive preparative HPLC systems, the final product was identified by HPLC and then analyzed by (1)H, (13)C, (1)H-(1)H COSY NMR spectrometry.
Chung
Young Soo
YS
Genechem Inc., 59-5 Jang-dong, Yuseong-gu, Daejeon 305-343, South Korea.
Kim
Dae Hee
DH
Seo
Won Min
WM
Lee
Hei Chan
HC
Liou
Kwangkyoung
K
Oh
Tae-Jin
TJ
Sohng
Jae Kyung
JK
D000595:Amino Acid Sequence; D003837:Deoxy Sugars / Q000096:biosynthesis* / Q000138:chemical synthesis; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D013171:Spores, Bacterial / Q000737:chemistry / Q000201:enzymology / Q000235:genetics; D013302:Streptomyces / Q000737:chemistry / Q000201:enzymology / Q000235:genetics; D013942:Thymine Nucleotides / Q000096:biosynthesis* / Q000138:chemical synthesis; D000637:Transaminases / Q000737:chemistry* / Q000235:genetics
D003837:Deoxy Sugars; D013942:Thymine Nucleotides; C456723:dTDP-4-amino-4,6-dideoxy-glucose; D000637:Transaminases; C050782:dTDP-4-keto-6-deoxyglucose aminotransferase