title: Efficient preparation of enantiopure D-phenylalanine through asymmetric resolution using immobilized phenylalanine ammonia-lyase from Rhodotorula glutinis JN-1 in a recirculating packed-bed reactor.
abstract: An efficient enzymatic process was developed to produce optically pure D-phenylalanine through asymmetric resolution of the racemic DL-phenylalanine using immobilized phenylalanine ammonia-lyase (RgPAL) from Rhodotorula glutinis JN-1. RgPAL was immobilized on a modified mesoporous silica support (MCM-41-NH-GA). The resulting MCM-41-NH-GA-RgPAL showed high activity and stability. The resolution efficiency using MCM-41-NH-GA-RgPAL in a recirculating packed-bed reactor (RPBR) was higher than that in a stirred-tank reactor. Under optimal operational conditions, the volumetric conversion rate of L-phenylalanine and the productivity of D-phenylalanine reached 96.7 mM h⁻¹ and 0.32 g L⁻¹ h⁻¹, respectively. The optical purity (eeD) of D-phenylalanine exceeded 99%. The RPBR ran continuously for 16 batches, the conversion ratio did not decrease. The reactor was scaled up 25-fold, and the productivity of D-phenylalanine (eeD>99%) in the scaled-up reactor reached 7.2 g L⁻¹ h⁻¹. These results suggest that the resolution process is an alternative method to produce highly pure D-phenylalanine.
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China; School of Biochemical Engineering, Anhui Polytechnic University, Wuhu, Anhui, China.
authors.forename: Longbao
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Huang
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Cui
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China.
authors.forename: Zhongmei
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China.
authors.forename: Ke
authors.initials: K
authors.lastname: Xiao
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China.
authors.forename: Zhemin
authors.initials: Z
authors.lastname: Zhou
mesh_terms: D000327:Adsorption; D055162:Biocatalysis; D005591:Chemical Fractionation / Q000379:methods; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000302:isolation & purification; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification; D007700:Kinetics; D010649:Phenylalanine / Q000737:chemistry*; D010650:Phenylalanine Ammonia-Lyase / Q000737:chemistry* / Q000302:isolation & purification; D012248:Rhodotorula / Q000737:chemistry* / Q000201:enzymology; D012822:Silicon Dioxide; D013237:Stereoisomerism
chemical_list: D004800:Enzymes, Immobilized; D005656:Fungal Proteins; D010649:Phenylalanine; D012822:Silicon Dioxide; D010650:Phenylalanine Ammonia-Lyase