Identification and characterization of the Rhizobium sp. strain GIN611 glycoside oxidoreductase resulting in the deglycosylation of ginsenosides.
Using enrichment culture, Rhizobium sp. strain GIN611 was isolated as having activity for deglycosylation of a ginsenoside, compound K (CK). The purified heterodimeric protein complex from Rhizobium sp. GIN611 consisted of two subunits with molecular masses of 63.5 kDa and 17.5 kDa. In the genome, the coding sequence for the small subunit was located right after the sequence for the large subunit, with one nucleotide overlapping. The large subunit showed CK oxidation activity, and the deglycosylation of compound K was performed via oxidation of ginsenoside glucose by glycoside oxidoreductase. Coexpression of the small subunit helped soluble expression of the large subunit in recombinant Escherichia coli. The purified large subunit also showed oxidation activity against other ginsenoside compounds, such as Rb1, Rb2, Rb3, Rc, F2, CK, Rh2, Re, F1, and the isoflavone daidzin, but at a much lower rate. When oxidized CK was extracted and incubated in phosphate buffer with or without enzyme, (S)-protopanaxadiol [PPD(S)] was detected in both cases, which suggests that deglycosylation of oxidized glucose is spontaneous.
School of Chemical and Biological Engineering, Seoul National University, Seoul, Republic of Korea.
Eun-Mi
EM
Kim
Juhan
J
Kim
Joo-Hyun
JH
Seo
Jun-Seong
JS
Park
Duck-Hee
DH
Kim
Byung-Gee
BG
Kim
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D004269:DNA, Bacterial / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D036145:Ginsenosides / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D005894:Panax / Q000382:microbiology; D010936:Plant Extracts / Q000737:chemistry / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism; D012231:Rhizobium / Q000201:enzymology*; D012502:Sapogenins / Q000737:chemistry / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D004269:DNA, Bacterial; D036145:Ginsenosides; D010936:Plant Extracts; D011994:Recombinant Proteins; D012502:Sapogenins; D010088:Oxidoreductases; C062916:protopanaxadiol