Ancestral Haloalkane Dehalogenases Show Robustness and Unique Substrate Specificity
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00159816%3A_____%2F17%3A00067125" target="_blank" >RIV/00159816:_____/17:00067125 - isvavai.cz</a>
Alternative codes found
RIV/00216224:14310/17:00095408
Result on the web
<a href="http://dx.doi.org/10.1002/cbic.201700197" target="_blank" >http://dx.doi.org/10.1002/cbic.201700197</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1002/cbic.201700197" target="_blank" >10.1002/cbic.201700197</a>
Alternative languages
Result language
angličtina
Original language name
Ancestral Haloalkane Dehalogenases Show Robustness and Unique Substrate Specificity
Original language description
Ancestral sequence reconstruction (ASR) represents a powerful approach for empirical testing structure-function relationships of diverse proteins. We employed ASR to predict sequences of five ancestral haloalkane dehalogenases (HLDs) from the HLD-II subfamily. Genes encoding the inferred ancestral sequences were synthesized and expressed in Escherichia coli, and the resurrected ancestral enzymes (AncHLD1-5) were experimentally characterized. Strikingly, the ancestral HLDs exhibited significantly enhanced thermodynamic stability compared to extant enzymes (Delta T-m up to 24 degrees C), as well as higher specific activities with preference for short multi-substituted halogenated substrates. Moreover, multivariate statistical analysis revealed a shift in the substrate specificity profiles of AncHLD1 and AncHLD2. This is extremely difficult to achieve by rational protein engineering. The study highlights that ASR is an efficient approach for the development of novel biocatalysts and robust templates for directed evolution.
Czech name
—
Czech description
—
Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
—
OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
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)
Others
Publication year
2017
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
ChemBioChem
ISSN
1439-4227
e-ISSN
—
Volume of the periodical
18
Issue of the periodical within the volume
14
Country of publishing house
DE - GERMANY
Number of pages
9
Pages from-to
1448-1456
UT code for WoS article
000405726100015
EID of the result in the Scopus database
—