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KMID : 0545120080180061081
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Journal of Microbiology and Biotechnology
2008 Volume.18 No. 6 p.1081 ~ p.1089
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Purification and Properties of a Novel β-Glucosidase, Hydrolyzing Ginsenoside Rb1 to CK, from Paecilomyces Bainier
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Yan Qin
Zhou Xin-Wen
Zhou Wei
Li Xing-Wei
Zhou Pei
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Abstract
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A novel ginsenoside-hydrolyzing β-glucosidase was purified from Paecilomyces Bainier sp. 229 by a combination of QSepharose FF, phenyl-Sepharose CL-4B, and CHT ceramic hydroxyapatite column chromatographies. The purified enzyme was a monomeric protein with a molecular mass estimated to be 115 kDa. The optimal enzyme activity was observed at pH 3.5 and 60oC. It was highly stable within pH 3-9 and at temperatures lower than 55oC. The enzyme was specific to β-glucoside. The order of enzyme activities against different types of β-glucosidic linkages was β-(1- 6)>β-(1-2)>β-(1-4). The enzyme converted ginsenoside Rb1 to CK specifically and efficiently. An 84.3% amount of ginsenoside Rb1, with an initial concentration of 2 mM, was converted into CK in 24 h by the enzyme at 45oC and pH 3.5. The hydrolysis pathway of ginsenoside Rb1 by the enzyme was Rb1→Rd→F2→CK. Five tryptic peptide fragments of the enzyme were identified by a newly developed de novo sequencing method of post-source decay (PSD) matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. By comparing the five identified peptide sequences with the NCBI database, this purified β-glucosidase proves to be a new protein that has not been reported before.
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KEYWORD
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Ginsenoside CK, ginsenoside-hydrolyzing β-glucosidase, purification, Paecilomyces Bainier
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