3TSY

4-Coumaroyl-CoA Ligase::Stilbene Synthase fusion protein

Summary for 3TSY

• Entry DOI: 10.2210/pdb3tsy/pdb
• Related: 1XES 1Z1E 3A9U 3A9V
• Descriptor: Fusion Protein 4-coumarate--CoA ligase 1, Resveratrol synthase (1 entity in total)
• Functional Keywords: fusion protein, ligase, transferase
• Biological source: Arabidopsis thaliana (mouse-ear cress,thale-cress, wine grape); More
• Total number of polymer chains: 1
• Total formula weight: 106631.47

Authors

Yi, H.,Jez, J.M. (deposition date: 2011-09-13, release date: 2011-12-14, Last modification date: 2024-02-28)

Primary citation

Wang, Y.,Yi, H.,Wang, M.,Yu, O.,Jez, J.M.
Structural and Kinetic Analysis of the Unnatural Fusion Protein 4-Coumaroyl-CoA Ligase::Stilbene Synthase.
J.Am.Chem.Soc., 133:20684-20687, 2011

PubMed Abstract: To increase the biochemical efficiency of biosynthetic systems, metabolic engineers have explored different approaches for organizing enzymes, including the generation of unnatural fusion proteins. Previous work aimed at improving the biosynthesis of resveratrol, a stilbene associated a range of health-promoting activities, in yeast used an unnatural engineered fusion protein of Arabidopsis thaliana (thale cress) 4-coumaroyl-CoA ligase (At4CL1) and Vitis vinifera (grape) stilbene synthase (VvSTS) to increase resveratrol levels 15-fold relative to yeast expressing the individual enzymes. Here we present the crystallographic and biochemical analysis of the 4CL::STS fusion protein. Determination of the X-ray crystal structure of 4CL::STS provides the first molecular view of an artificial didomain adenylation/ketosynthase fusion protein. Comparison of the steady-state kinetic properties of At4CL1, VvSTS, and 4CL::STS demonstrates that the fusion protein improves catalytic efficiency of either reaction less than 3-fold. Structural and kinetic analysis suggests that colocalization of the two enzyme active sites within 70 Å of each other provides the basis for enhanced in vivo synthesis of resveratrol.

PubMed: 22129213
DOI: 10.1021/ja2085993

Experimental method

X-RAY DIFFRACTION (3.1 Å)