Medizinische Universität Graz - Research portal
Selected Publication:
SHR Neuro Cancer Cardio Lipid Metab Microb
Drienovská, I; Gajdoš, M; Kindler, A; Takhtehchian, M; Darnhofer, B; Birner-Gruenberger, R; Dörr, M; Bornscheuer, UT; Kourist, R.
Folding assessment of incorporation of non-canonical amino acids facilitates expansion of functional group diversity for enzyme engineering.
Chemistry. 2020;
Doi: 10.1002/chem.202002077
[OPEN ACCESS]
Web of Science
PubMed
FullText
FullText_MUG
- Co-authors Med Uni Graz
- Birner-Grünberger Ruth
- Darnhofer Barbara
- Altmetrics:
- Dimensions Citations:
- Plum Analytics:
- Scite (citation analytics):
- Abstract:
- Protein design is limited by the diversity of functional groups provided by the canonical protein "building blocks". Incorporating non-canonical amino acids (ncAAs) into enzymes enables a dramatic expansion of their catalytic features. For this, quick identification of fully translated and correctly folded variants is crucial. Herein, we report the engineering of the enantioselectivity of an esterase utilizing several ncAAs. Key for the identification of active and soluble protein variants was the use of the split-GFP method, which is crucial as it allows simple determination of the expression levels of enzyme variants with ncAA incorporations by fluorescence. Several identified variants led to improved enantioselectivity or even inverted enantiopreference in the kinetic resolution of ethyl 3-phenylbutyrate. © 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
- Find related publications in this database (Keywords)
- biocatalysis
- enzyme expression
- noncanonical amino acids
- protein engineering
- pseudomonas fluorescensesterase