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

Kód výsledku v 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>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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

Název v anglickém jazyce

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

Popis výsledku anglicky

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

Druh

D - Stať ve sborníku

CEP obor

CE - Biochemie

OECD FORD obor

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Projekt

<a href="/cs/project/LC06010" target="_blank" >LC06010: Centrum biokatalýzy a biotransformací</a><br>

Návaznosti

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

Rok uplatnění

2006

Kód důvěrnosti údajů

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

Název statě ve sborníku

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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Počet stran výsledku

1

Strana od-do

160

Název nakladatele

Institute of Microbiology ASCR

Místo vydání

Praha

Místo konání akce

Praha

Datum konání akce

1. 1. 2006

Typ akce podle státní příslušnosti

WRD - Celosvětová akce

Kód UT WoS článku

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