Protein engineering of haloalkane dehalogenase LinB: reconstruction of active site and modification of entrance tunnel

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F06%3A00017059" target="_blank" >RIV/00216224:14310/06:00017059 - 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

Protein engineering of haloalkane dehalogenase LinB: reconstruction of active site and modification of entrance tunnel

Original language description

Haloalkane dehalogenase LinB is an enzyme isolated from lindan degrading bacterium Sphingobium japonicum UT26. LinBs 3D structure [1], catalytic properties and substrate specificity are known and well studied. Thanks to these facts LinB is great target for protein engineering experiments. Firts experiment, reconstruction of active site, was based on 68% sequence identity with ORF rv2579 from Mycobacterium tuberculosis H37Rv genome. The homology model of protein Rv2579 was compared with the 3D structureof LinB. This analysis revealed that 6 out of 19 amino acid residues which form an active site and entrance tunnel are different in LinB and Rv2579. The 6 different amino acids were cumulatively mutated in LinB. Final six-fold mutant was presumed to haveactive site and entrance tunnel of Rv2579 and exhibited dehalogenase activity with the haloalkanes tested, confirming that Rv2579 is a member of the haloalkane dehalogenase family. Consequently the M. tuberculosis gene rv2579 was cloned

Czech name

Proteinové inženýrství haloalkán dehalogenázy LinB: rekonstrukce aktivního místa a modifikace vstupního tunelu.

Czech description

Haloalkán dehalogenáza LinB je enzym izolovaný z bakterie Sphingobium japonicum UT26, která je schopna degradovat lindan. Struktura LinB, katalytické vlastnosti a substrátová specifita jsou dobře znamy. proto je LinB vhodným kandidátem pro proteinové experimenty.

Type

D - Article in proceedings

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)

Publication year

2006

Confidentiality

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

Article name in the collection

Book of Abstracts, 10th International Symposium of the Genetics of Industrial Microorganisms

ISBN

80-239-7323-1

ISSN

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e-ISSN

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Number of pages

1

Pages from-to

160

Publisher name

Institute of Microbiology ASCR

Place of publication

Praha

Event location

Praha

Event date

Jan 1, 2006

Type of event by nationality

WRD - Celosvětová akce

UT code for WoS article

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