Enzymatic transformation of Rg3 to Rh2 using newly isolated Fusarium proliferatum ECU2042Enzymatic transformation of Rg3 to Rh2 using newly isolated Fusarium proliferatum ECU2042.docVIP

Abstract  Journal of Molecular Catalysis B: Enzymatic 38 (2006) 113–118 Enzymatic 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 proliferatum  1. 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