Engineering nonphosphorylative metabolism to synthesize mesaconate from lignocellulosic sugars in Escherichia coli
| Autor: | Yi Shu Tai, Wenqin Bai, Kevin J. Fox, Kechun Zhang, Pooja Jambunathan, Jilong Wang, Jingyu Wang |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
030106 microbiology Pentoses Pentose Bioengineering Biology Pentose phosphate pathway medicine.disease_cause Applied Microbiology and Biotechnology Lignin Metabolic engineering 03 medical and health sciences Fumarates medicine Escherichia coli Phosphorylation chemistry.chemical_classification Xylose Escherichia coli Proteins Maleates Metabolism Arabinose Biosynthetic Pathways Citric acid cycle Enzyme Genetic Enhancement chemistry Biochemistry Metabolic Engineering Yield (chemistry) Metabolic Networks and Pathways Biotechnology |
| Zdroj: | Metabolic engineering. 38 |
| ISSN: | 1096-7184 |
| Popis: | Dicarboxylic acids are attractive biosynthetic targets due to their broad applications and their challenging manufacturing process from fossil fuel feedstock. Mesaconate is a branched, unsaturated dicarboxylic acid that can be used as a co-monomer to produce hydrogels and fire-retardant materials. In this study, we engineered nonphosphorylative metabolism to produce mesaconate from d-xylose and l-arabinose. This nonphosphorylative metabolism is orthogonal to the intrinsic pentose metabolism in Escherichia coli and has fewer enzymatic steps and a higher theoretical yield to TCA cycle intermediates than the pentose phosphate pathway. Here mesaconate production was enabled from the d-xylose pathway and the l-arabinose pathway. To enhance the transportation of d-xylose and l-arabinose, pentose transporters were examined. We identified the pentose/proton symporter, AraE, as the most effective transporter for both d-xylose and l-arabinose in mesaconate production process. Further production optimization was achieved by operon screening and metabolic engineering. These efforts led to the engineered strains that produced 12.5g/l and 13.2g/l mesaconate after 48h from 20g/l of d-xylose and l-arabinose, respectively. Finally, the engineered strain overexpressing both l-arabinose and d-xylose operons produced 14.7g/l mesaconate from a 1:1 d-xylose and l-arabinose mixture with a yield of 85% of the theoretical maximum. (0.87g/g). This work demonstrates an effective system that converts pentoses into a value-added chemical, mesaconate, with promising titer, rate, and yield. |
| Databáze: | OpenAIRE |
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