Metabolic engineering of Escherichia coli for quinolinic acid production by assembling L-aspartate oxidase and quinolinate synthase as an enzyme complex |
| |
Affiliation: | 1. Institute of Plant Sciences Paris-Saclay (IPS2), CNRS, INRA, Univ. Paris-Sud, Univ. Evry, Univ. Paris-Diderot, Université Paris-Saclay, Bâtiment 630, Rue Noetzlin, 91192, Gif-sur-Yvette cedex, France;2. Plateforme Métabolisme Métabolome, Institute of Plant Sciences Paris-Saclay (IPS2), CNRS, INRA, Univ. Paris-Sud, Univ. Evry, Univ. Paris-Diderot, Université Paris-Saclay, Bâtiment 630, Rue Noetzlin, 91192, Gif-sur-Yvette cedex, France;3. UMR 1332 Biologie du Fruit et Pathologie, INRA, 33883, Villenave d’Ornon, France;1. Department of BioSciences, Rice University, Houston, 77005 TX, USA;2. Department of Bioengineering, Rice University, Houston, 77005 TX, USA;3. Department of Chemical and Biomolecular Engineering, Rice University, Houston, 77005 TX, USA |
| |
Abstract: | Quinolinic acid (QA) is a key intermediate of nicotinic acid (Niacin) which is an essential human nutrient and widely used in food and pharmaceutical industries. In this study, a quinolinic acid producer was constructed by employing comprehensive engineering strategies. Firstly, the quinolinic acid production was improved by deactivation of NadC (to block the consumption pathway), NadR (to eliminate the repression of L-aspartate oxidase and quinolinate synthase), and PtsG (to slow the glucose utilization rate and achieve a more balanced metabolism, and also to increase the availability of the precursor phosphoenolpyruvate). Further modifications to enhance quinolinic acid production were investigated by increasing the oxaloacetate pool through overproduction of phosphoenolpyruvate carboxylase and deactivation of acetate-producing pathway enzymes. Moreover, quinolinic acid production was accelerated by assembling NadB and NadA as an enzyme complex with the help of peptide-peptide interaction peptides RIAD and RIDD, which resulted in up to 3.7 g/L quinolinic acid being produced from 40 g/L glucose in shake-flask cultures. A quinolinic acid producer was constructed in this study, and these results lay a foundation for further engineering of microbial cell factories to efficiently produce quinolinic acid and subsequently convert this product to nicotinic acid for industrial applications. |
| |
Keywords: | Quinolinic acid Peptide-peptide interaction Enzyme complex Metabolon Metabolite channeling Metabolic engineering |
本文献已被 ScienceDirect 等数据库收录! |
|