title: A high-throughput adrenaline test for the exploration of the catalytic potential of halohydrin dehalogenases in epoxide ring-opening reactions.
abstract: The principle of the adrenaline test for enzymes is based on the quantification of periodate-sensitive reaction products with adrenaline to produce a chromogenic compound adrenochrome that can be easily detected. Here, a rapid whole-cell -based adrenaline assay for the activity measurement of halohydrin dehalogenases (HHDHs) in nucleophile-mediated epoxide ring-opening reactions is presented. The assay was validated using two types of model reactions (glycidol with nucleophiles and nitrite with epoxides). Moreover, the reliability of the assay was confirmed by gas chromatography analysis. Our results demonstrated that the developed assay is efficient in both library screening and the evaluation of catalytic diversity and specificity of HHDHs. Thus, the assay represents a valuable tool in the evolution of HHDHs for its industrial applications. Moreover, the adrenaline test exhibits a great potential for enzyme assay and could be easily adopted for other suitable enzymes.
authors.affiliation: School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, People's Republic of China.
authors.forename: Lixia
authors.initials: L
authors.lastname: Tang
authors.affiliation: School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, People's Republic of China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, People's Republic of China.
authors.forename: Rongxiang
authors.initials: R
authors.lastname: Jiang
authors.affiliation: School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, People's Republic of China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, People's Republic of China.
authors.forename: Kai
authors.initials: K
authors.lastname: Zheng
authors.affiliation: School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, People's Republic of China.
authors.forename: Huayu
authors.initials: H
authors.lastname: Zheng
mesh_terms: D000323:Adrenochrome / Q000737:chemistry; D016960:Agrobacterium tumefaciens / Q000201:enzymology* / Q000235:genetics; D001173:Arthrobacter / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D002384:Catalysis; D004837:Epinephrine / Q000737:chemistry*; D006867:Hydrolases / Q000737:chemistry* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D000323:Adrenochrome; D006867:Hydrolases; C430848:halohydrin dehalogenase; D004837:Epinephrine
keywords: activity assay; halohydrin dehalogenases; β-substituted alcohols