Catalysis of Thermostable Alcohol Dehydrogenase Improved by Engineering the Microenvironment through Fusion with Supercharged Proteins.
enzymatic microenvironment
protein complexation
steady-state enzyme kinetics
substrate channeling
supercharged proteins
Journal
Chembiochem : a European journal of chemical biology
ISSN: 1439-7633
Titre abrégé: Chembiochem
Pays: Germany
ID NLM: 100937360
Informations de publication
Date de publication:
15 07 2019
15 07 2019
Historique:
received:
30
01
2019
revised:
09
03
2019
pubmed:
13
3
2019
medline:
22
7
2020
entrez:
13
3
2019
Statut:
ppublish
Résumé
The enzymatic microenvironment can impact biocatalytic activity; however, these effects can be difficult to investigate as mutations and fusions can introduce multiple variables and overlapping effects. The fusion of a supercharged protein is a potentially facile means to alter the enzymatic microenvironment. We have investigated complexes made between a thermostable alcohol dehydrogenase (AdhD) and superfolding green fluorescent protein (sfGFP) mutants with extreme surface charges. Three charged sfGFP variants, -30, 0, and +36 were covalently attached to AdhD through the SpyCatcher/SpyTag system. Specific rates for the NAD
Identifiants
pubmed: 30859665
doi: 10.1002/cbic.201900066
doi:
Substances chimiques
Archaeal Proteins
0
Butylene Glycols
0
Peptides
0
SpyCatcher peptide
0
SpyTag peptide
0
Green Fluorescent Proteins
147336-22-9
2,3-butylene glycol
45427ZB5IJ
Alcohol Dehydrogenase
EC 1.1.1.1
Types de publication
Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
1827-1837Informations de copyright
© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.