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To increase activity and stability of HHDH, 14 amino acid residues around the active site and substrate-binding pocket based on the structural analysis and molecular docking were selected as targets for site-directed mutagenesis. The studies showed that the mutant HHDH (Mut-HHDH) enzyme had a more accessible substrate-binding pocket than the wild-type HHDH (Wt-HHDH). Molecular docking revealed that the distance between the substrate and active site was closer in mutant which improved the catalytic activity. The expressed Wt-HHDH and Mut-HHDH were purified and characterized using 1,3-dichloro-2-propanol (1,3-DCP) as substrates. The specific activity of the mutant was enhanced 26-fold and the value of k \u00a0cat was 18.4-fold as compared to the Wt-HHDH, respectively. The Mut-HHDH showed threefold extension of half-life at 45\u00a0\u00b0C than that of Wt-HHDH. Therefore it is possible to add 1,3-DCP concentration up to 100\u00a0mM and epichlorohydrin (ECH) was produced at a relatively high conversion and yield (59.6\u00a0%) using Mut-HHDH as catalyst. This Mut-HHDH could be a potential candidate for the upscale production of ECH.<\/jats:p>","DOI":"10.1007\/s10295-014-1443-2","type":"journal-article","created":{"date-parts":[[2014,4,28]],"date-time":"2014-04-28T11:01:25Z","timestamp":1398682885000},"page":"1145-1158","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["Expression, characterization, and improvement of a newly cloned halohydrin dehalogenase from<i>Agrobacterium tumefaciens<\/i>and its application in production of epichlorohydrin"],"prefix":"10.1093","volume":"41","author":[{"given":"Zhi-Qiang","family":"Liu","sequence":"first","affiliation":[{"name":"grid.413273.0 0000000105748737 Institute of Bioengineering Zhejiang University of Technology 310014 Hangzhou Zhejiang People\u2019s Republic of 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China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yin-Chu","family":"Shen","sequence":"additional","affiliation":[{"name":"grid.413273.0 0000000105748737 Institute of Bioengineering Zhejiang University of Technology 310014 Hangzhou Zhejiang People\u2019s Republic of China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2014,7,1]]},"reference":[{"issue":"1","key":"2021033103313659800_CR1","doi-asserted-by":"publisher","first-page":"491","DOI":"10.1146\/annurev.micro.51.1.491","article-title":"Synthesis of enantiopure epoxides through biocatalytic approaches","volume":"51","author":"Archelas","year":"1997","journal-title":"Annu Rev Microbiol"},{"issue":"7","key":"2021033103313659800_CR2","doi-asserted-by":"publisher","first-page":"568","DOI":"10.1016\/S0141-0229(97)00254-8","article-title":"Biochemical characterization of a haloalcohol dehalogenase from Arthrobacter erithii 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