title: Covalent immobilization of Agrobacterium radiobacter epoxide hydrolase on ethylenediamine functionalised epoxy supports for biocatalytical synthesis of (R)-epichlorohydrin.
abstract: OBJECTIVE
  To improve the operational stability and reusability of an epoxide hydrolase (EH) for the biosynthesis of optically active epoxides.
  
  
  RESULTS
  A covalently immobilization strategy was employed to improve the stability of Agrobacterium radiobacter EH by using ethylenediamine (EDA)-functionalised epoxy resin LX-1000EP as carrier. Under the optimal conditions, the activity recovery of immobilized enzyme was 72 % and the specific activity was 634 U/g. Immobilized EH exhibited significantly enhanced thermal stability with a half-life of more than 6.8-fold at 50 °C than that of the free enzyme. A gram-scale kinetic resolution of (R,S)-epichlorohydrin using immobilized preparation as biocatalyst was performed and (R)-epichlorohydrin was obtained with 35 % yield and 99 % enantiomeric excess. The immobilized EH showed good operational stability and even after six reactions, it retained >85 % of the initial activity.
  
  
  CONCLUSIONS
  The operational stability and recyclability of immobilized EH on an EDA-functionalized epoxy supports demonstrated its potential for producing (R)-epichlorohydrin.
authors.affiliation: Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.
authors.forename: Shu-Ping
authors.initials: SP
authors.lastname: Zou
authors.affiliation: Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.
authors.forename: Zhi-Cai
authors.initials: ZC
authors.lastname: Wang
authors.affiliation: Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China.
authors.forename: Chao
authors.initials: C
authors.lastname: Qin
authors.affiliation: Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, People's Republic of China. zhengyg@zjut.edu.cn.
authors.forename: Yu-Guo
authors.initials: YG
authors.lastname: Zheng
mesh_terms: D016960:Agrobacterium tumefaciens / Q000201:enzymology*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D004811:Epichlorohydrin / Q000737:chemistry / Q000378:metabolism*; D004851:Epoxide Hydrolases / Q000737:chemistry / Q000378:metabolism*; D005029:Ethylenediamines / Q000737:chemistry*
chemical_list: D004800:Enzymes, Immobilized; D005029:Ethylenediamines; D004811:Epichlorohydrin; C031234:ethylenediamine; D004851:Epoxide Hydrolases
keywords: (R)-epichlorohydrin; Biocatalytical synthesis; Covalent immobilization; Epoxide hydrolas; Epoxy supports; Ethylenediamine