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Screening of glucosidase and identification of its reaction pathway using mass spectrometers

초록

영어

In this study, screening of microorganism which has hydrolysis activity to compound K was attempted. As a result, one microorganism was found. Screened microorganism was analyzed by 16s rDNA sequencing and proved to be Sphingobacterium multivorum. For screening of target glucosidase from microorganism, we tried to purify of enzyme and find the N-terminal
sequence. Target enzyme was purified and sequenced. But N-terminal of enzyme was blocked. Due to N-terminal modification of the enzyme, internal sequences after tryptic digestion were obtained using mass spectrometers, and the information was used for gene cloning. S. multivorum is showed activity to other ginseng saponin such as Rb1 as well as Compound K. Ginsenoside Rb1 hydrolysis reaction pathway can be also predicted using mass spectrometry. MS/MS analysis results suggests that the screened enzyme catalyzes sequential glucose
hydrolysis reaction to produce the various intermediates and final product from Rb1, i.e., Rb1Gp-XVIIGp-LXXV Compound KPPD(S).

저자정보

  • Eun-Mi Kim School of Chemical and Biological Engineering, Seoul National University
  • Duck-Hee Kim R & D Center, Amore-Pacific Corporation, Kyounggi-do, Korea.
  • Byung Gee Kim School of Chemical and Biological Engineering, Seoul National University

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