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

Abstract

e Here, we isolated and characterized a new ginsenoside-transforming ␤-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 ( ؊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 ␤-glucosidase of glycoside hydrolase family 3.

Identification and Characterization of a Ginsenoside-Transforming β-Glucosidase from Pseudonocardia sp. Gsoil 1536 and Its Application for Enhanced Production of Minor Ginsenoside Rg2(S)

“…Gsoil 1536 was isolated. In addition, the near-neutral optimal pH and mild optimal temperature of BglPC28 are similar to those of other ginsenoside-hydrolyzing GH3 from bacteria [28], [29], [35], [46], [47]. Although the optimum temperature of BglPC28 for pNPG is 37°C, the ginsenoside-conversion reaction occurred at 30°C for extension of stable transformation activity.…”

Production of the Rare Ginsenoside Rh2-MIX (20(S)-Rh2, 20(R)-Rh2, Rk2, and Rh3) by Enzymatic Conversion Combined with Acid Treatment and Evaluation of Its Anti-Cancer Activity

“…To date, many studies have attempted to find suitable enzymes to develop a bioprocess to transform major ginsenosides to minor ginsenoside, such as for F2 [21], 20(S)-Rg3 [20], C-K [27], Rh1 [23], 20(S)-Rg2 [22], and F1 [6]. For example, β-glucosidase cloned from Microbacterium esteraromaticum could effectively transform ginsenosides Rb1 and Rd into 20(S)-Rg3 by hydrolysis of the outer and inner glucoses at the C20 position [23]. However, there is no report of the production of ginsenoside Rh2 by transformation from major ginsenosides directly because of the difficulty of cleaving the three glucose moieties specifically (outer and inner glucoses at the C3 position, and the outer glucose at the C20 position).…”

“…presented a simple enzymatic digestion experiment, without further scale-up or process engineering. To date, several recombinant ginsenoside hydrolyzing enzymes have been constructed to produce ginsenosides 20(S)-Rg3 [20], F2 [21], 20(S)-Rg2 [22], 20(S)-Rh1 [23], and F1 [6] on a large scale However, there is no report of the production of Rh2 using bioprocess engineering.…”

Characterization of a Novel Ginsenoside MT1 Produced by an Enzymatic Transrhamnosylation of Protopanaxatriol-Type Ginsenosides Re

“…The standard forms of various ginsenosides (Rg 1 and Re) used in the present study were purchased from Sigma Co., Ltd. (Louis, MO, USA). F 1 , Rg 2 (S), Rh 1 (S) and PPT were prepared as described in our previous study [22]. The other chemical reagents used were at least extra or better in quality than pure grade.…”