Sunday , 3 March 2024
Home Science Kinetic compartmentalization by unnatural reaction for itaconate production – Nature Communications
Science

Kinetic compartmentalization by unnatural reaction for itaconate production – Nature Communications

In particular, endogenous 2-methylcitrate dehydratase, PrpD19 is successfully engineered that originally converts 2-methylcitrate into 2-methyl-cis-aconitate, to catalyze the conversion of citrate into cis-aconitate. PrpD is evolutionarily engineered to switch its specificity from 2-methylcitrate to citrate by computational simulation design and high-throughput screening20,21,22. Based on the non-natural enzyme, an unnatural itaconate production pathway is constructed that is kinetically isolated from the TCA cycle (Fig. 1). Using our approach, we obtain a significantly increased itaconate yield of up to 10.6-fold compared to the parental strain at 46.0% of the theoretical maximum14. This implementation of kinetic compartmentalization in bacteria allows overcoming the limitation of being a spatially inseparable host by introducing an unnatural biochemical reaction able to efficiently separate competitive reactions. This method shows potential for kinetic compartmentalization in prokaryotes, considering the substantial number of promiscuous enzymes that can be engineered using this approach.

 

Continue reading your article with a FrontPage News Subscription

Already a Subscriber? Sign In  

OR
VIEW SUBSCRIPTION OPTIONS
Support independent journalism by subscribing