Detailed Information on Publication Record
2017
Ancestral Haloalkane Dehalogenases Show Robustness and Unique Substrate Specificity
BABKOVÁ, Petra, Eva ŠEBESTOVÁ, Jan BREZOVSKÝ, Radka CHALOUPKOVÁ, Jiří DAMBORSKÝ et. al.Basic information
Original name
Ancestral Haloalkane Dehalogenases Show Robustness and Unique Substrate Specificity
Authors
BABKOVÁ, Petra (203 Czech Republic, belonging to the institution), Eva ŠEBESTOVÁ (203 Czech Republic, belonging to the institution), Jan BREZOVSKÝ (203 Czech Republic, belonging to the institution), Radka CHALOUPKOVÁ (203 Czech Republic, belonging to the institution) and Jiří DAMBORSKÝ (203 Czech Republic, guarantor, belonging to the institution)
Edition
CHEMBIOCHEM, 2017, 1439-4227
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10608 Biochemistry and molecular biology
Country of publisher
Germany
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
Impact factor
Impact factor: 2.774
RIV identification code
RIV/00216224:14310/17:00095408
Organization unit
Faculty of Science
UT WoS
000405726100015
Keywords in English
ancestral sequence reconstruction; haloalkane dehalogenase; protein engineering; robustness; substrate specificity
Změněno: 28/3/2018 15:36, Ing. Nicole Zrilić
Abstract
V originále
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 (DTm up to 24 8C), 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.
Links
| GA16-06096S, research and development project |
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| GA16-24223S, research and development project |
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| LM2015047, research and development project |
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| LM2015051, research and development project |
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| LM2015055, research and development project |
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| LO1214, research and development project |
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