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Canvasing the Substrate-Binding Pockets of the Wax Ester Synthase.
Natalia Calixto Mancipe1, Kalene M Mulliner1, Mary H Plunkett2
1Department of Bioproducts and Biosystems Engineering, University of Minnesota, St. Paul, Minnesota 55108, United States.
Biochemistry
|May 4, 2022
Summary
Bacterial wax ester synthase (WS/DGAT) enzymes are key to producing valuable lipids and biofuels. This study uses structural insights to modify enzyme substrate specificity for improved biotechnological applications.
Area of Science:
- Biochemistry
- Enzymology
- Synthetic Biology
Background:
- Wax ester synthase/acyl-coenzyme A:diacylglycerol acyltransferase (WS/DGAT) enzymes synthesize wax esters and triglycerides in bacteria.
- These enzymes exhibit broad substrate specificity, accepting various alcohols, diglycerides, and fatty acyl-CoAs, indicating significant biotechnological potential for lipid and biofuel production.
Purpose of the Study:
- To leverage the crystal structure of *Marinobacter aquaeolei* VT8 WS/DGAT (Maqu_0168) to understand enzyme architecture.
- To investigate the role of amino acids in substrate-binding pockets and their impact on substrate profiles.
- To develop methods for probing residues that influence fatty acyl-CoA selectivity.
Main Methods:
- Structural analysis of *Marinobacter aquaeolei* VT8 WS/DGAT.
- Site-directed mutagenesis of amino acid residues within substrate-binding pockets.
- Characterization of altered substrate profiles and fatty acyl-CoA selectivity.
Main Results:
- Identification of key amino acid residues influencing the substrate specificity of WS/DGAT.
- Demonstration that specific residue exchanges alter the enzyme's ability to accept different alcohols and fatty acyl-CoAs.
- Development of a novel approach to investigate fatty acyl-CoA selectivity determinants.
Conclusions:
- The study provides a structural blueprint for understanding WS/DGAT substrate binding and catalysis.
- Enzyme engineering strategies based on structural insights can tailor WS/DGAT for specific biotechnological applications.
- Further research can optimize these enzymes for enhanced lipid and biofuel biosynthesis.
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