In:
Applied and Environmental Microbiology, American Society for Microbiology, Vol. 79, No. 19 ( 2013-10), p. 5788-5798
Abstract:
Here, we isolated and characterized a new ginsenoside-transforming β-glucosidase (BglQM) from Mucilaginibacter sp. strain QM49 that shows biotransformation activity for various major ginsenosides. The gene responsible for this activity, bglQM , consists of 2,346 bp and is predicted to encode 781 amino acid residues. This enzyme has a molecular mass of 85.6 kDa. Sequence analysis of BglQM revealed that it could be classified into glycoside hydrolase family 3. The enzyme was overexpressed in Escherichia coli BL21(DE3) using a maltose binding protein (MBP)-fused pMAL-c2x vector system containing the tobacco etch virus (TEV) proteolytic cleavage site. Overexpressed recombinant BglQM could efficiently transform the protopanaxatriol-type ginsenosides Re and Rg 1 into ( S )-Rg 2 and ( S )-Rh 1 , respectively, by hydrolyzing one glucose moiety attached to the C-20 position at pH 8.0 and 30°C. The K m values for p -nitrophenyl-β- d -glucopyranoside, Re, and Rg 1 were 37.0 ± 0.4 μM and 3.22 ± 0.15 and 1.48 ± 0.09 mM, respectively, and the V max values were 33.4 ± 0.6 μmol min −1 mg −1 of protein and 19.2 ± 0.2 and 28.8 ± 0.27 nmol min −1 mg −1 of protein, respectively. A crude protopanaxatriol-type ginsenoside mixture (PPTGM) was treated with BglQM, followed by silica column purification, to produce ( S )-Rh 1 and ( S )-Rg 2 at chromatographic purities of 98% ± 0.5% and 97% ± 1.2%, respectively. This is the first report of gram-scale production of ( S )-Rh 1 and ( S )-Rg 2 from PPTGM using a novel ginsenoside-transforming β-glucosidase of glycoside hydrolase family 3.
Type of Medium:
Online Resource
ISSN:
0099-2240
,
1098-5336
DOI:
10.1128/AEM.01150-13
Language:
English
Publisher:
American Society for Microbiology
Publication Date:
2013
detail.hit.zdb_id:
223011-2
detail.hit.zdb_id:
1478346-0
SSG:
12
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