Cyclization specificity in terpene synthases by residue interaction networks

Lead Research Organisation: John Innes Centre
Department Name: Contracts Office

Abstract

Enzymes are essential to life and critical for industrial biotechnology efforts aimed at producing everyday pharmaceuticals, biofuels, and fine chemicals. A complete understanding of enzyme function will enable us to fully exploit enzymes to successfully adapt to an ever changing and uncertain world. We are continually making major strides in our understanding of enzyme function through focusing on the active site where the chemical reactions occur. However, it is well documented in directed evolution experiments that outer tier mutations (regions of the enzyme structure distant from the active site) greatly enhance catalytic activity. Although outer tiers of protein structure clearly exert profound influence on enzyme function, this remains poorly understood; our current knowledge rests largely on retrospective structure-based analysis while limited investigations have been conducted.

This project seeks to deliver new fundamental knowledge into enzyme function by exploiting graph theory to identify distant residues that impact catalytic activities and establishing their linkage to the active site through residue interaction networks. Rich data sets of biochemical data, terpene products and sequences will seed computational studies to develop physical models to understand how residue interaction networks transmit mutational information through protein structure to modulate catalysis. These advances will enable formulation of general approaches for protein engineering to create libraries that are enriched with catalytically robust and functionally diverse enzymes that will ultimately reduce the resources associated with screening efforts. Developing these approaches using terpene synthase will generate novel biocatalysts for industrial biotechnology to support our growing need for therapeutics, flavors, fragrances, and other fine chemicals.

Publications

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