Efficient production of S-(+)-2-chlorophenylglycine by immobilized penicillin G acylase in a recirculating packed bed reactor |
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| Affiliation: | 1. Institute of Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China;2. Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou 310014, China;1. Chemistry Department, Roudehen Branch, Islamic Azad University, Roudehen, Iran;2. Chemistry Department, Shahid Beheshti University, G. C., Tehran, Evin, Iran;1. Chemistry Dept., Hunter College, City University Of New York, 695 Park Ave, New York, NY 10065, USA;2. CUNY Graduate Center, 365 5th Avenue, New York, NY 10016, USA;1. Department of Chemical Engineering, National Chung Hsing University, 250 Kuo Kuang Road, Taichung, 40227, Taiwan, ROC;2. Biotechnology Center, National Chung Hsing University, 250 Kuo Kuang Road, Taichung, 40227, Taiwan, ROC;1. Karlsruhe Institute of Technology, Institute for Neutron Physics and Reactor Technology (INR), Hermann-von-Helmholtz-Platz 1, D-76344 Eggenstein-Leopoldshafen, Germany;2. Karlsruhe Institute of Technology, Institute for Applied Materials-Applied Materials Physics (IAM-AWP), Hermann-von-Helmholtz-Platz 1, D-76344 Eggenstein-Leopoldshafen, Germany;3. Institut de Radioprotection et de Sûreté Nucléaire (IRSN), Nuclear Safety Division/Safety Research/Severe Accident Department, Saint Paul Lez Durance 13115, France |
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| Abstract: | (S)-(+)-2-Chlorophenylglycine 1 is an important intermediate in the synthesis of Clopidogrel. A recirculating packed bed reactor (RPBR) was constructed for efficient production of (S)-1 by kinetic resolution of racemic N-phenylacetyl-2- chlorophenylglycine 2 using immobilized penicillin G acylase (PGA). The immobilized PGA exhibited maximum activity at 50 °C and pH 8.0 with (R,S)-2 as substrate. The kinetic constants (Km and vmax) of immobilized PGA were calculated to be 20.61 mM and 83.2 mM/min/g, respectively. The substrate displayed inhibitory effect on immobilized PGA with inhibition constant of 221.23 mM. The immobilized PGA showed a strict enantiospecificity for substrate at different temperature, pH and substrate concentration examined. The performance and productivity of RPBR were evaluated by several critical parameters, including immobilized PGA load, substrate feeding rate, height to diameter ratio and so on. The kinetic resolution process shows higher initial reaction rate and conversion by recycling 100 mL of substrate solution (80 mM) through RPBRs packed with 6.0 g immobilized PGA with a feeding rate of 1.5 mL/min while the H/D ratio was 4.0. The immobilized PGA-catalyzed kinetic resolution of (R,S)-2 was successfully operated in the RPBR for 60 batches, with an average productivity of 1.2 g/L/h for (S)-1 in high optical purity (>97% enantiomeric excess) in semi-continuous operation. The residual (R)-2 can be easily racemized and then used as substrate. |
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| Keywords: | S-(+)-2-Chlorophenylglycine Immobilized penicillin G acylase Biocatalysis Bioconversion Packed bed bioreactors Optimization |
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