A reverse glyoxylate shunt to build a non-native route from C4 to C2 in Escherichia coli |
| |
| Affiliation: | 1. Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, CA 90095, USA;2. Biomedical Engineering IDP, University of California, Los Angeles, CA 90095, USA];1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China;2. National Engineering Laboratory for Cereal Fermentation Technology (NELCF), Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China;3. School of Pharmaceutical Science, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China;1. Laboratory for Bioinformatics, Graduate School of Systems Biosciences, Kyushu University, 804 Westwing, 3-1-1 Maidashi, Higashi-ku, Fukuoka, Japan;2. Laboratory of Microbial Physiology, Research Faculty of Agriculture, Hokkaido University, Kita 9 Nishi 9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan;1. Bioprocess Engineering, Wageningen University, Wageningen UR PO Box 16, 6700AA, Wageningen, The Netherlands;2. Biobased Products, Wageningen University and Research Centre, Wageningen, The Netherlands |
| |
| Abstract: | Most central metabolic pathways such as glycolysis, fatty acid synthesis, and the TCA cycle have complementary pathways that run in the reverse direction to allow flexible storage and utilization of resources. However, the glyoxylate shunt, which allows for the synthesis of four-carbon TCA cycle intermediates from acetyl-CoA, has not been found to be reversible to date. As a result, glucose can only be converted to acetyl-CoA via the decarboxylation of the three-carbon molecule pyruvate in heterotrophs. A reverse glyoxylate shunt (rGS) could be extended into a pathway that converts C4 carboxylates into two molecules of acetyl-CoA without loss of CO2. Here, as a proof of concept, we engineered in Escherichia coli such a pathway to convert malate and succinate to oxaloacetate and two molecules of acetyl-CoA. We introduced ATP-coupled heterologous enzymes at the thermodynamically unfavorable steps to drive the pathway in the desired direction. This synthetic pathway in essence reverses the glyoxylate shunt at the expense of ATP. When integrated with central metabolism, this pathway has the potential to increase the carbon yield of acetate and biofuels from many carbon sources in heterotrophic microorganisms, and could be the basis of novel carbon fixation cycles. |
| |
| Keywords: | Glyoxylate shunt Central metabolism Non-native pathways Acetyl-coA biosynthesis Reversibility Primary metabolism |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
