Design and Evolution of Haloalkane Dehalogenase with Enhanced Conversion of 1,2,3-Trichloropropane by Modification of Access Tunnels

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F09%3A00032484" target="_blank" >RIV/00216224:14310/09:00032484 - isvavai.cz</a>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

Design and Evolution of Haloalkane Dehalogenase with Enhanced Conversion of 1,2,3-Trichloropropane by Modification of Access Tunnels

Original language description

Engineering enzymes to degrade anthropogenic compounds efficiently is challenging. We obtained Rhodococcus rhodochrous haloalkane dehalogenase mutants with up to 32-fold higher activity than wild type toward the toxic, recalcitrant anthropogenic compound1,2,3-trichloropropane (TCP) using a new strategy. We identified key residues in access tunnels connecting the buried active site with bulk solvent by rational design and randomized them by directed evolution. The most active mutant has large aromatic residues at two out of three randomized positions and two positions modified by site-directed mutagenesis. These changes apparently enhance activity with TCP by decreasing accessibility of the active site for water molecules, thereby promoting activated complex formation. Kinetic analyses confirmed that the mutations improved carbon-halogen bond cleavage and shifted the rate-limiting step to the release of products.

Czech name

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Czech description

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Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

CE - Biochemistry

OECD FORD branch

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Project

<a href="/en/project/LC06010" target="_blank" >LC06010: Center of Biocatalysis and Biotransformation</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>Z - Vyzkumny zamer (s odkazem do CEZ)

Publication year

2009

Confidentiality

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

Name of the periodical

Nature Chemical Biology

ISSN

1552-4450

e-ISSN

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Volume of the periodical

5

Issue of the periodical within the volume

xx

Country of publishing house

US - UNITED STATES

Number of pages

7

Pages from-to

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UT code for WoS article

000270039900010

EID of the result in the Scopus database

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