title: Immobilization of dehydrogenase onto epoxy-functionalized nanoparticles for synthesis of (R)-mandelic acid.
abstract: Epoxy functionalized magnetic Fe3O4@SiO2 nanoparticles were successfully prepared and characterized by Fourier-transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM). The prepared nanoparticles were used for immobilization of alcohol dehydrogenase (ADH) from Saccharomyces cerevisiae by covalent attachment. The optimal immobilization conditions were obtained as follows: enzyme/support 4.49mg/g, pH 8.0, buffer concentration 0.05M, time 12h and temperature 30°C. Under these conditions, a high immobilization yield and efficiency of above 92% were obtained after the optimization. Broad pH tolerance and high thermostability were achieved by the immobilization. The immobilized ADH retained about 84% initial activity after five cycles. Kinetic parameters Vmax and Km of free and immobilized ADH were determined as 56.72μM/min, 44.27μM/min and 11.54mM, 31.32mM, respectively. (R)-mandelic acid synthesis with the immobilized ADH was carried out, and the yield of (R)-mandelic acid was as high as 64%. These results indicate that the ADH immobilized onto epoxy-functionalized nanoparticles is an efficient and simple way for preparation of stable ADH, and the immobilized ADH has potential applications in the production of (R)-mandelic acid.
authors.affiliation: School of Pharmacy, Jiangsu University, Zhenjiang 212013, People's Republic of China.
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Jiang
authors.affiliation: School of Pharmacy, Jiangsu University, Zhenjiang 212013, People's Republic of China.
authors.forename: Ting-Ting
authors.initials: TT
authors.lastname: Lu
authors.affiliation: School of Pharmacy, Jiangsu University, Zhenjiang 212013, People's Republic of China.
authors.forename: Cai-Hong
authors.initials: CH
authors.lastname: Liu
authors.affiliation: School of Pharmacy, Jiangsu University, Zhenjiang 212013, People's Republic of China.
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Ling
authors.affiliation: School of Pharmacy, Jiangsu University, Zhenjiang 212013, People's Republic of China.
authors.forename: Meng-Yao
authors.initials: MY
authors.lastname: Zhuang
authors.affiliation: United Pharmaceutical Institute of Jiangsu University and Shandong Tianzhilvye Biotechnology Co., Ltd., Jiangsu University, Zhenjiang 212013, People's Republic of China.
authors.forename: Jiu-Xun
authors.initials: JX
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Jiangsu University, Zhenjiang 212013, People's Republic of China; United Pharmaceutical Institute of Jiangsu University and Shandong Tianzhilvye Biotechnology Co., Ltd., Jiangsu University, Zhenjiang 212013, People's Republic of China. Electronic address: zhangyewang@ujs.edu.cn.
authors.forename: Ye-Wang
authors.initials: YW
authors.lastname: Zhang
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification; D055162:Biocatalysis; D002021:Buffers; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000302:isolation & purification; D004853:Epoxy Resins / Q000737:chemistry; D052203:Ferrosoferric Oxide / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification; D006038:Glyoxylates / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D058185:Magnetite Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D008333:Mandelic Acids / Q000138:chemical synthesis* / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000201:enzymology; D012822:Silicon Dioxide / Q000737:chemistry; D013237:Stereoisomerism; D013696:Temperature
chemical_list: D002021:Buffers; D004800:Enzymes, Immobilized; D004853:Epoxy Resins; D005656:Fungal Proteins; D006038:Glyoxylates; D058185:Magnetite Nanoparticles; D008333:Mandelic Acids; C012482:phenylglyoxylic acid; D012822:Silicon Dioxide; D000426:Alcohol Dehydrogenase; C037938:mandelic acid; D052203:Ferrosoferric Oxide
keywords: (R)-Mandelic acid; Alcohol dehydrogenase; Covalent attachment; Immobilization; Magnetic nanoparticles