Identification and Characterization of a NovelTerrabacter ginsenosidimutanssp. nov. β-Glucosidase That Transforms Ginsenoside Rb1 into the Rare Gypenosides XVII and LXXV

Microbial β-Glucosidases: Screening, Characterization, Cloning and Applications

“…Generally fungal β-glucosidase exhibits optimal activity at pH of 4.0-6.0 [59, 69,86,92,93], nevertheless β-glucosidase with optimal pH of 2.4 has been reported from T. cylindrosporum Syzx4 [68] and that with optimal pH of 9 and 10 has been reported from Acremonium murorun LPSC 927 and Chaetomium globosum, respectively [94,95]. Bacterial β-glucosidase exhibits optimal activity at pH of 6-7 [44,87,96,97], although those with pH optima of 5, 8 and 10 has been reported from Caldicellulosiruptor saccharolyticus [98], Bacillus halodurans [99], Klebsiella pneumoniae [100], respectively. Yeast β-glucosidase reported from A. pullulans and Candida peltata exhibited optimal activity at pH 5 [80,101] and that from Kluyveromyces marxianus showed optimal activity at pH 5.5 [102].…”

Identification and Characterization of a Mucilaginibacter sp. Strain QM49 β-Glucosidase and Its Use in the Production of the Pharmaceutically Active Minor Ginsenosides ( S )-Rh 1 and ( S )-Rg 2

“…strain QM49 was isolated. In addition, the near-neutral optimal pH and mild optimal temperature of BglQM are similar to those of other ginsenoside-hydrolyzing family 3 ␤-glucosidases from bacteria (23,32,56).…”

“…The results, which are summarized in Table 3, showed that BglQM was maximally active against PNPGlc followed by ONPGlc but had little effect on various other PNP-and ONP-glycosides, with the exception of PNP-␣-L-arabinofuranoside and PNP-␤-D-fucopyranoside. The specific activity of BglQM against PNP-␣-L-arabinofuranoside is not seen in ginsenoside-transforming glycoside hydrolase family 3 members from Terrabacter ginsenosidimutans (23) and Microbacterium esteraromaticum (32). However, BglQM cannot hydrolyze the arabinofuranoside at the outer position of the C-20 of the ginsenoside Rc, perhaps due to structural differences between the molecules.…”

“…When Rb 1 , Rd, Re, and Rg 1 (1.0 mg/ml) were used as substrates, they were biotransformed within 10 min by 10 mg/ml of crude recombinant MBP-BglQM. Several other ginsenoside-hydrolyzing recombinant enzymes have been reported, but most were only able to hydrolyze the outer or inner glucose moiety at the C-3 position of the aglycon (23,24,26). One previously described enzyme from Microbacterium esteraromaticum KCTC 12040BP (32) had the same ability as BglQM, but the authors conducted only a simple enzymatic characterization without further scale up or process engineering.…”

“…However, these approaches produce nonspecific stereoisomeric mixtures of rare ginsenosides that have low yields and are very difficult to separate. As an alternative, enzymatic methods have been explored as a way to more specifically convert major ginsenosides into pharmacologically active rare ginsenosides (23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)52). Furthermore, methods have been developed to efficiently separate a PPT-type ginsenoside mixture (PPTGM) and a PPD-type ginsenoside mixture (PPDGM) from ginseng extracts (34,35).…”