Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Torres-Salas, P; Bernal, V; López-Gallego, F; Martínez-Crespo, J; Sánchez-Murcia, PA; Barrera, V; Morales-Jiménez, R; García-Sánchez, A; Mañas-Fernández, A; Seoane, JM; Sagrera Polo, M; Miranda, JD; Calvo, J; Huertas, S; Torres, JL; Alcalde-Bascones, A; González-Barrera, S; Gago, F; Morreale, A; González-Barroso, MDM.
Engineering Erg10 Thiolase from Saccharomyces cerevisiae as a Synthetic Toolkit for the Production of Branched-Chain Alcohols.
Biochemistry. 2018; 57(8):1338-1348
Doi: 10.1021/acs.biochem.7b01186
Web of Science
PubMed
FullText
FullText_MUG
- Co-Autor*innen der Med Uni Graz
- Sánchez Murcia Pedro Alejandro
- Altmetrics:
- Dimensions Citations:
- Plum Analytics:
- Scite (citation analytics):
- Abstract:
- Thiolases catalyze the condensation of acyl-CoA thioesters through the Claisen condensation reaction. The best described enzymes usually yield linear condensation products. Using a combined computational/experimental approach, and guided by structural information, we have studied the potential of thiolases to synthesize branched compounds. We have identified a bulky residue located at the active site that blocks proper accommodation of substrates longer than acetyl-CoA. Amino acid replacements at such a position exert effects on the activity and product selectivity of the enzymes that are highly dependent on a protein scaffold. Among the set of five thiolases studied, Erg10 thiolase from Saccharomyces cerevisiae showed no acetyl-CoA/butyryl-CoA branched condensation activity, but variants at position F293 resulted the most active and selective biocatalysts for this reaction. This is the first time that a thiolase has been engineered to synthesize branched compounds. These novel enzymes enrich the toolbox of combinatorial (bio)chemistry, paving the way for manufacturing a variety of α-substituted synthons. As a proof of concept, we have engineered Clostridium's 1-butanol pathway to obtain 2-ethyl-1-butanol, an alcohol that is interesting as a branched model compound.
- Find related publications in this database (using NLM MeSH Indexing)
- Acetyl-CoA C-Acetyltransferase - chemistry
- Acetyl-CoA C-Acetyltransferase - genetics
- Acetyl-CoA C-Acetyltransferase - metabolism
- Acyl Coenzyme A - metabolism
- Catalytic Domain -
- Hexanols - metabolism
- Metabolic Networks and Pathways -
- Models, Molecular -
- Protein Engineering - methods
- Saccharomyces cerevisiae - chemistry
- Saccharomyces cerevisiae - genetics
- Saccharomyces cerevisiae - metabolism
- Saccharomyces cerevisiae Proteins - chemistry
- Saccharomyces cerevisiae Proteins - genetics
- Saccharomyces cerevisiae Proteins - metabolism