caa a22 4500 605504172 CHVBK 20210128100614.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s00253-015-6446-z doi (NATIONALLICENCE)springer-10.1007/s00253-015-6446-z Enzymatic transformation of vina-ginsenoside R7 to rare notoginsenoside ST-4 using a new recombinant glycoside hydrolase from Herpetosiphon aurantiacus [Elektronische Daten] [Ru-Feng Wang, Ming-Min Zheng, Yue-De Cao, Hao Li, Chun-Xiu Li, Jian-He Xu, Zheng-Tao Wang] An eco-friendly and convenient preparation method for notoginsenoside ST-4 has been established by completely transforming vina-ginsenoside R7 using a recombinant glycosidase hydrolyzing enzyme (HaGH03) from Herpetosiphon aurantiacus. This enzyme specifically hydrolyzed the glucose at the C-20 position but not the external xylose or two inner glucoses at position C-3. Protein sequence BLAST revealed that HaGH03, composed of 749 amino acids and presumptively listed as a member of the family 3 glycoside hydrolases, has highest identity (48%) identity with a thermostable β-glucosidase B, which was not known of any functions for ginsenoside transformation. The steady state kinetic parameters for purified HaGH03 measured against p-nitrophenyl β-D-glucopyranoside and vina-ginsenoside R7 were K M = 5.67 ± 0.24μM and 0.59 ± 0.23mM, and k cat = 69.2 ± 0.31/s and 2.15 ± 0.46/min, respectively. HaGH03 converted 2.5mg/mL of vina-ginsenoside R7 to ST-4 with a molar yield of 100% and a space-time yield of 104mg/L/h in optimized conditions. These results underscore that HaGH03 has much potential for the effective preparation of target ginsenosides possessing valuable pharmacological activities. This is the first report identifying an enzyme that has the ability to transform vina-ginsenoside R7 and provides an approach to preparing rare notoginsenoside ST-4. Springer-Verlag Berlin Heidelberg, 2015 Biotransformation nationallicence Notoginsenoside ST-4 nationallicence Vina -ginsenoside R7 nationallicence Family 3 glycoside hydrolase nationallicence Herpetosiphon aurantiacus nationallicence Heterologous expression nationallicence Wang Ru-Feng Department of Pharmacognosy, China Pharmaceutical University, 210038, Nanjing, People's Republic of China aut Zheng Ming-Min State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, People's Republic of China aut Cao Yue-De State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, People's Republic of China aut Li Hao State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, People's Republic of China aut Li Chun-Xiu State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, People's Republic of China aut Xu Jian-He State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, People's Republic of China aut Wang Zheng-Tao Department of Pharmacognosy, China Pharmaceutical University, 210038, Nanjing, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/8(2015-04-01), 3433-3442 0175-7598 99:8<3433 2015 99 253 https://doi.org/10.1007/s00253-015-6446-z text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6446-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wang Ru-Feng Department of Pharmacognosy, China Pharmaceutical University, 210038, Nanjing, People's Republic of China aut NATIONALLICENCE 700 1- Zheng Ming-Min State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Cao Yue-De State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Li Hao State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Li Chun-Xiu State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Xu Jian-He State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Wang Zheng-Tao Department of Pharmacognosy, China Pharmaceutical University, 210038, Nanjing, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/8(2015-04-01), 3433-3442 0175-7598 99:8<3433 2015 99 253