Transhalogenation Catalysed by Haloalkane Dehalogenases Engineered to Stop Natural Pathway at Intermediate

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F19%3A00071029" target="_blank" >RIV/00159816:_____/19:00071029 - isvavai.cz</a>

Alternative codes found

RIV/00216224:14310/19:00113347

Result on the web

<a href="https://onlinelibrary.wiley.com/doi/pdf/10.1002/adsc.201900132" target="_blank" >https://onlinelibrary.wiley.com/doi/pdf/10.1002/adsc.201900132</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/adsc.201900132" target="_blank" >10.1002/adsc.201900132</a>

Result language

angličtina

Original language name

Transhalogenation Catalysed by Haloalkane Dehalogenases Engineered to Stop Natural Pathway at Intermediate

Original language description

Haloalkane dehalogenases (HLDs) are alpha/beta-hydrolases that convert halogenated compounds to their corresponding alcohols. The overall kinetic mechanism proceeds via four steps: (i) binding of halogenated substrate, (ii) bimolecular nucleophilic substitution (S(N)2) leading to the cleavage of a carbon-halogen bond and the formation of an alkyl-enzyme intermediate, (iii) nucleophilic addition of a water molecule resulting in the hydrolysis of the intermediate to the corresponding alcohol and (iv) release of the reaction products - an alcohol, a halide ion and a proton. Although, the overall reaction has been reported as irreversible, several kinetic evidences from previous studies suggest the reversibility of the first S(N)2 chemical step. To study this phenomenon, we have engineered HLDs to stop the catalytic cycle at the stage of the alkyl-enzyme intermediate. The ability of the intermediate to exchange halides was confirmed by a stopped-flow fluorescence binding analysis. Finally, the transhalogenation reaction was confirmed with several HLDs and 2,3-dichloropropene in the presence of a high concentration of iodide. The formation of the transhalogenation product 3-iodo-2-chloropropene catalysed by five mutant HLDs was identified by gas chromatography coupled with mass spectrometry. Hereby we demonstrated the reversibility of the cleavage of the carbon-halogen bond by HLDs resulting in a transhalogenation. After optimization, the transhalogenation reaction can possibly find its use in biocatalytic applications. Enabling this reaction by strategically engineering the enzyme to stop at an intermediate in the catalytic cycle that is synthetically more useful than the product of the natural pathway is a novel concept.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10406 - Analytical chemistry

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2019

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

ADVANCED SYNTHESIS &amp; CATALYSIS

ISSN

1615-4150

e-ISSN

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Volume of the periodical

361

Issue of the periodical within the volume

11

Country of publishing house

US - UNITED STATES

Number of pages

5

Pages from-to

2438-2442

UT code for WoS article

000471070400007

EID of the result in the Scopus database

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