ENZYMATIC BIOPROCESSING OF PECTIN-RICH AGRICULTURAL BYPRODUCTS

The major goal for this project isto develop a value-adding in vitro enzymatic conversion technology for the conversion of pectic monosaccharides into specialty chemicals. This enzymatic strategy can enable rapid bioprocessing with reduced handling and capex requirements compared to conventional approaches such as chemical synthesis or fermentation. Implementation of this biomanufacturing process can increase overall crop revenue. This offers reduced volatility in crop value, financial sustainability for farmers, and construction of rural-located fermentation facilities to create high paying technical jobs.The overarching measurable objective for towards this goal is to produce and purify an enzyme for the conversion of the pectin sugar, D-galacturonic acid into a high value chemical intermediate. However, the current enzyme activity is limiting to production. Therefore the goal for this project are to improve enzyme activity.To achieve this overall objective, we have two main technical objectives:Screen enzyme homologs for high activty on pectin hydrolysate. We will rapidly synthesize and screen a variety of enzyme homologs using in vitro expression for high activity on D-galacturonic acid in pectin hydrolysate.High throughput enzyme engineering for improved activity. Using the best enzyme sequence from Objective 1, we will perform iterative PCR mutagenesis on this sequence and identify high activity mutants. Top hits from each round will be characterized for activity under production conditions and used as template for subsequent rounds of mutagensis.