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Enzymtic transformation of Rg3 to Rh2 using newly isolated Fusarium proliferatum ECU2042.doc

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Abstract Journal of Molecular Catalysis B: Enzymatic 38 (2006) 113–118Enzymatic transformation of ginsenoside Rg3to Rh2using newly isolated Fusarium proliferatum ECU2042 Jin-Huan Sua, Jian-He Xua,?, Wen-Ya Lub, Guo-Qiang Linb,? a Laboratory of Biocatalysis and Bioprocessing, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China b Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, China Received 13 September 2005; received in revised form 1 December 2005; accepted 10 December 2005 A new soil isolate, Fusarium proliferatum ECU2042, was found capable of producing a ginsenoside Rg3hydrolase (a special -glucosidase) which is useful for production of a very potent antitumor agent Rh2. By investigating its fermentation progress, the optimal time for -glucosidase biosynthesis was determined as 72 h. The crude -glucosidase was partially puri?ed by 49.4-fold from the cell-free extract with an activity yield of 13%. The catalytic performance of the partially puri?ed -glucosidase was examined, giving optima of pH and temperature at pH 5.0–6.0 and 50 ?C. It was found that the F. proliferatum -glucosidase can also hydrolyze cellobiose, but not active towards a frequently used arti?cial substrate p-nitrophenyl -d-glucopyranoside (pNPG). In a lab-scale preparation, the bio-hydrolytic reaction was carried out at 50 ?C for 24 h in 50 ml NaAc/HAc buffer (100 mM, pH 5.0) containing 50 mg Rg3and 12 mg lyophilized crude enzyme extract, giving the ginsenoside Rh2in 60.3% conversion. ? 2005 Elsevier B.V. All rights reserved. Keywords: Ginseng saponin; Ginsenoside Rg3; Ginsenoside Rh2 ; -Glucosidase; Fusarium proliferatum1. Introduction Saponins, glycosides with steroids or triterpenes as agly- cons, are an important class of physiologically active compounds occurring in many herbs. In recent years, the sugar chains of the saponins were found to be closely related to the biological acti