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Purification and characterization of a ginsenoside Rb(1)-hydrolyzing β-glucosidase from Aspergillus niger KCCM 11239.

Chang KH, Jo MN, Kim KT, Paik HD - Int J Mol Sci (2012)

Bottom Line: In the specificity tests, the enzyme was found to be active against ginsenoside Rb(1), but showed very low levels of activity against Rb(2), Rc, Rd, Re, and Rg(1).The enzyme hydrolyzed the 20-C,β-(1→6)-glucoside of ginsenoside Rb(1) to generate ginsenoside Rd and Rg(3), and hydrolyzed 3-C,β-(1→2)-glucoside to generate F(2).The properties of the enzyme indicate that it could be a useful tool in biotransformation applications in the ginseng industry, as well as in the development of novel drug compounds.

View Article: PubMed Central - PubMed

Affiliation: Division of Animal Life Science, Konkuk University, Seoul 143-701, Korea; E-Mails: khchang80@cj.net (K.H.C); ulul55@naver.com (M.N.J.).

ABSTRACT
Rb(1)-hydrolyzing β-glucosidase from Aspergillus niger KCCM 11239 was studied to develop a bioconversion process for minor ginsenosides. The specific activity of the purified enzyme was 46.5 times greater than that of the crude enzyme. The molecular weight of the native enzyme was estimated to be approximately 123 kDa. The optimal pH of the purified enzyme was pH 4.0, and the enzyme proved highly stable over a pH range of 5.0-10.0. The optimal temperature was 70 °C, and the enzyme became unstable at temperatures above 60 °C. The enzyme was inhibited by Cu(2+), Mg(2+), Co(2+), and acetic acid (10 mM). In the specificity tests, the enzyme was found to be active against ginsenoside Rb(1), but showed very low levels of activity against Rb(2), Rc, Rd, Re, and Rg(1). The enzyme hydrolyzed the 20-C,β-(1→6)-glucoside of ginsenoside Rb(1) to generate ginsenoside Rd and Rg(3), and hydrolyzed 3-C,β-(1→2)-glucoside to generate F(2). The properties of the enzyme indicate that it could be a useful tool in biotransformation applications in the ginseng industry, as well as in the development of novel drug compounds.

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Related in: MedlinePlus

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme produced by Aspergillus niger KCCM 11239. M, protein marker; lane 1, supernatant fraction; lane 2, ammonium sulphate precipitation (30%–90%); lane 3, Sephadex G-100; lane 4, DEAE Sephadex.
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f4-ijms-13-12140: Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme produced by Aspergillus niger KCCM 11239. M, protein marker; lane 1, supernatant fraction; lane 2, ammonium sulphate precipitation (30%–90%); lane 3, Sephadex G-100; lane 4, DEAE Sephadex.

Mentions: The purified β-glucosidase appeared to be homogeneous according to the criteria of SDS-PAGE, as shown in Figure 4. The molecular weight of the purified β-glucosidase was estimated at approximately 123 kDa, based on calculations of its mobility using standard calibration proteins. The β-glucosidase from A. niger KCCM 11239 differs from those of other β-glucosidases generated by A. niger reported thus far, which were 46, 110, and 240 kDa [16–18].


Purification and characterization of a ginsenoside Rb(1)-hydrolyzing β-glucosidase from Aspergillus niger KCCM 11239.

Chang KH, Jo MN, Kim KT, Paik HD - Int J Mol Sci (2012)

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme produced by Aspergillus niger KCCM 11239. M, protein marker; lane 1, supernatant fraction; lane 2, ammonium sulphate precipitation (30%–90%); lane 3, Sephadex G-100; lane 4, DEAE Sephadex.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3472798&req=5

f4-ijms-13-12140: Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzyme produced by Aspergillus niger KCCM 11239. M, protein marker; lane 1, supernatant fraction; lane 2, ammonium sulphate precipitation (30%–90%); lane 3, Sephadex G-100; lane 4, DEAE Sephadex.
Mentions: The purified β-glucosidase appeared to be homogeneous according to the criteria of SDS-PAGE, as shown in Figure 4. The molecular weight of the purified β-glucosidase was estimated at approximately 123 kDa, based on calculations of its mobility using standard calibration proteins. The β-glucosidase from A. niger KCCM 11239 differs from those of other β-glucosidases generated by A. niger reported thus far, which were 46, 110, and 240 kDa [16–18].

Bottom Line: In the specificity tests, the enzyme was found to be active against ginsenoside Rb(1), but showed very low levels of activity against Rb(2), Rc, Rd, Re, and Rg(1).The enzyme hydrolyzed the 20-C,β-(1→6)-glucoside of ginsenoside Rb(1) to generate ginsenoside Rd and Rg(3), and hydrolyzed 3-C,β-(1→2)-glucoside to generate F(2).The properties of the enzyme indicate that it could be a useful tool in biotransformation applications in the ginseng industry, as well as in the development of novel drug compounds.

View Article: PubMed Central - PubMed

Affiliation: Division of Animal Life Science, Konkuk University, Seoul 143-701, Korea; E-Mails: khchang80@cj.net (K.H.C); ulul55@naver.com (M.N.J.).

ABSTRACT
Rb(1)-hydrolyzing β-glucosidase from Aspergillus niger KCCM 11239 was studied to develop a bioconversion process for minor ginsenosides. The specific activity of the purified enzyme was 46.5 times greater than that of the crude enzyme. The molecular weight of the native enzyme was estimated to be approximately 123 kDa. The optimal pH of the purified enzyme was pH 4.0, and the enzyme proved highly stable over a pH range of 5.0-10.0. The optimal temperature was 70 °C, and the enzyme became unstable at temperatures above 60 °C. The enzyme was inhibited by Cu(2+), Mg(2+), Co(2+), and acetic acid (10 mM). In the specificity tests, the enzyme was found to be active against ginsenoside Rb(1), but showed very low levels of activity against Rb(2), Rc, Rd, Re, and Rg(1). The enzyme hydrolyzed the 20-C,β-(1→6)-glucoside of ginsenoside Rb(1) to generate ginsenoside Rd and Rg(3), and hydrolyzed 3-C,β-(1→2)-glucoside to generate F(2). The properties of the enzyme indicate that it could be a useful tool in biotransformation applications in the ginseng industry, as well as in the development of novel drug compounds.

Show MeSH
Related in: MedlinePlus