Purification Method Improvement and Characterization of a Novel Ginsenoside-Hydrolyzing β-Glucosidase from<i>Paecilomyces Bainier</i>sp. 229

Yan, Qin; Zhou, Wei; Li, Xingwei; Feng, Meiqing; Zhou, Pei

doi:10.1271/bbb.70425

Cited by 53 publications

(32 citation statements)

References 24 publications

(21 reference statements)

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“…The specific activities of -glucosidase from P. bainier sp. 229 for Rd and Rb 1 have been reported to be 129 and 57 mmol/min/mg respectively, 20) 3-and 6-fold lower than that of -glycosidase from S. solfataricus (387 and 349 mmol/min/mg respectively). a All the substrates were reagent-grade, and were used at 1 mg/ml.…”

Section: Substrate Specificity and Biotransformation Of Ginsenosidesmentioning

confidence: 88%

Ginsenoside Compound K Production from Ginseng Root Extract by a Thermostable β-Glycosidase fromSulfolobus solfataricus

Noh

Son

Kim

et al. 2009

Bioscience, Biotechnology, and Biochemistry

116

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Section: Substrate Specificity and Biotransformation Of Ginsenosidesmentioning

confidence: 88%

Ginsenoside Compound K Production from Ginseng Root Extract by a Thermostable β-Glycosidase fromSulfolobus solfataricus

Noh

Son

Kim

et al. 2009

Bioscience, Biotechnology, and Biochemistry

116

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“…Extract in the Presence and Absence of b b-Glycosidase from S. acidocaldarius Because rare ginsenosides can be produced by the hydrolysis of sugar moieties from major ginsenosides using ginsenoside-hydrolyzing b-glycosidases, [15][16][17][18] the above reaction was performed using a thermostable b-glycosidase from the hyperthermophilic bacterium S. acidocaldarius and an extract prepared from ginseng root at pH 5.5 and 85°C for 24 h. The ginsenosides in the ginseng root extract were analyzed by HPLC using a C 18 column. Four typical major ginsenosides were detected with the same retention times as Rb 1 , Rb 2 , Rc, and Rd (Fig.…”

Section: Hplc Analysis Of the Ginsenosides In Ginseng Rootmentioning

confidence: 99%

Production of the Rare Ginsenosides Compound K, Compound Y, and Compound Mc by a Thermostable .BETA.-Glycosidase from Sulfolobus acidocaldarius

Noh

2009

Biological & Pharmaceutical Bulletin

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Panax ginseng has been used in traditional medicine to strengthen immunity, provide nutrition, and reduce fatigue. The anticancer, anti-inflammatory, and anti-proliferative effects of P. ginseng are attributed to ginsenosides found in the plant. [1][2][3] Pharmaceutically active ginsenosides present at low concentrations or absent in ginseng root are referred to as rare ginsenosides. These rare ginsenosides, including compound K, compound Y, and compound Mc can be produced by the hydrolysis of sugar moieties from the major ginsenosides Rb 1 , Rb 2 , Rc, and Rd, which account for more than 50% of the total ginsenosides. ) has yet to be reported. Compound K production has been achieved from ginseng root extract by enzymatic methods using Pectinex, b-galactosidase from Aspergillus oryzae, and b-glycosidase from Sulfolobus solfataricus.9-11) Compound K, compound Y, and compound Mc were produced using a crude extract from A. niger, while compound K and compound Mc were produced by fermentation from Fusarium sacchari.12,13) The fungi used in the production of the rare ginsenosides exhibited low productivity, and the enzymes involved were unknown. b-Glycosidase from S. solfataricus produced mainly compound K but not compound Y due to its high level of hydrolytic activity toward the sugar groups in the ginsenosides. To produce the rare ginsenosides compound K, compound Y, and compound Mc more efficiently, the discovery of a new enzyme with different hydrolytic activity toward these sugar groups is necessary.In this study, the ginsenosides produced from the major ginsenosides Rb 1 , Rb 2 , Rc, and Rd by a thermostable b-glycosidase from S. acidocaldarius were identified and their production was attempted using ginseng root extract and reagent-grade Rb 1 , Rb 2 , and Rc. MATERIALS AND METHODSMaterials Regent-grade Rb 1 , Rb 2 , Rc, and Rd were purchased from BTGin (Chungnam, Korea). Compound K was kindly provided by Professor Dong-Hyun Kim of Kyunghee University. Ginseng root extract was prepared by the previously reported method. 4)Bacterial Strains, Plasmid, and Culture Conditions The genomic DNA from S. acidocaldarius DSMZ 639 (DSMZ, Brauschweig, Germany), Escherichia coli ER2566 (New Englands Biolabs, Herfordshire, U.K.), and pTrc99a plasmid (Pharmacia, Piscataway, NJ, U.S.A.) were used as a source of b-glycosidase gene, a host, and a vector for expression, respectively. S. acidocaldarius was grown anaerobically at 70°C on Sulfolobus medium (DSM media formulation No. 88) for 5 d. The recombinant E. coli for protein expression was cultivated in a 2-l flask containing 500 ml of Luria-Bertani (LB) medium and 50 mg/ml of ampicillin at 37°C with agitation at 200 rpm until the optical density of bacteria reached 0.8 at 600 nm. Isopropyl-b-D-thiogalactopyranoside (IPTG) was added to the culture medium at a final concentration of 0.1 mM, and the enzyme was induced and expressed at 16°C for 12 h.Cloning of b b-Glycosidase Gene from S. acidocaldarius The genomic DNA from S. acidocaldarius was extracted using the genomic DNA ext...

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“…The molecular weight of the purified Bgp1 was 87.5 kDa, as determined by SDS-PAGE showing lower than previous reported ginsenosidehydrolyzing β-glucosidase (102 kDa) isolated from Paecilomyces Bainier sp. 229 (Yan et al 2008a). The enzyme had optimal activity at pH 7.0 higher than that of ginsenoside-hydrolyzing β-glucosidase (pH 3.5) isolated from Paecilomyces Bainier sp.…”

Section: Discussionmentioning

confidence: 95%

“…The enzyme had optimal activity at pH 7.0 higher than that of ginsenoside-hydrolyzing β-glucosidase (pH 3.5) isolated from Paecilomyces Bainier sp. 229 (Yan et al 2008a). The optimal temperature for Bgp1 activity was 40°C which was lower than the optimal temperatures for ginsenoside-hydrolyzing β-glucosidases (55°C) from Paecilomyces Bainier sp.…”

Section: Discussionmentioning

confidence: 99%

“…The optimal temperature for Bgp1 activity was 40°C which was lower than the optimal temperatures for ginsenoside-hydrolyzing β-glucosidases (55°C) from Paecilomyces Bainier sp. 229 and Sulfolobus acidocaldarius (85°C) (Yan et al 2008a; Theoretically, four glucose moieties attached to the ginsenoside Rb1 could be available for hydrolysis by Bgp1, namely the outer and inner glucose moieties attached at positions C-3 and C-20. Based on an analysis of the hydrolysis products of ginsenoside Rb1, Bgp1 acted on not C-3 position but C-20 position, hydrolyzed two glucose moieties attached to the C-20 position of Rb1, effectively converted into Rg3.…”

Section: Discussionmentioning

confidence: 99%

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Enzymatic biotransformation of ginsenoside Rb1 to 20(S)-Rg3 by recombinant β-glucosidase from Microbacterium esteraromaticum

Quan

Min

Yang

et al. 2012

Appl Microbiol Biotechnol

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Microbacterium esteraromaticum was isolated from ginseng field. The β-glucosidase gene (bgp1) from M. esteraromaticum was cloned and expressed in Escherichia coli BL21 (DE3). The bgp1 gene consists of 2,496 bp encoding 831 amino acids which have homology to the glycosyl hydrolase family 3 protein domain. The recombinant β-glucosidase enzyme (Bgp1) was purified and characterized. The molecular mass of purified Bgp1 was 87.5 kDa, as determined by SDS-PAGE. Using 0.1 mg ml(-1) enzyme in 20 mM sodium phosphate buffer at 37°C and pH 7.0, 1.0 mg ml(-1) ginsenoside Rb1 was transformed into 0.444 mg ml(-1) ginsenoside Rg3 within 6 h. The Bgp1 sequentially hydrolyzed the outer and inner glucose attached to the C-20 position of ginsenosides Rb1. Bgp1 hydrolyzed the ginsenoside Rb1 along the following pathway: Rb1 → Rd → 20(S)-Rg3. This is the first report of the biotransformation of ginsenoside Rb1 to ginsenoside 20(S)-Rg3 using the recombinant β-glucosidase.

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Purification Method Improvement and Characterization of a Novel Ginsenoside-Hydrolyzing β-Glucosidase fromPaecilomyces Bainiersp. 229

Cited by 53 publications

References 24 publications

Ginsenoside Compound K Production from Ginseng Root Extract by a Thermostable β-Glycosidase fromSulfolobus solfataricus

Ginsenoside Compound K Production from Ginseng Root Extract by a Thermostable β-Glycosidase fromSulfolobus solfataricus

Production of the Rare Ginsenosides Compound K, Compound Y, and Compound Mc by a Thermostable .BETA.-Glycosidase from Sulfolobus acidocaldarius

Enzymatic biotransformation of ginsenoside Rb1 to 20(S)-Rg3 by recombinant β-glucosidase from Microbacterium esteraromaticum

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