Manipulating pyruvate to acetyl-CoA conversion in Escherichia coli for anaerobic succinate biosynthesis from glucose with the yield close to the stoichiometric maximum
| Autor: | Debabov Vladimir G, A. A. Morzhakova, Alexandra Yu. Skorokhodova, Andrey Yu. Gulevich |
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| Rok vydání: | 2015 |
| Předmět: |
Succinic Acid
Bioengineering Biology medicine.disease_cause Applied Microbiology and Biotechnology chemistry.chemical_compound Acetyl Coenzyme A Acetyltransferases Pyruvic Acid medicine Escherichia coli Anaerobiosis Escherichia coli Proteins Acetyl-CoA General Medicine Pyruvate dehydrogenase complex Citric acid cycle Glucose Biochemistry chemistry Metabolic Engineering Succinic acid Yield (chemistry) Fermentation Pyruvic acid Metabolic Networks and Pathways Biotechnology |
| Zdroj: | Journal of biotechnology. 214 |
| ISSN: | 1873-4863 |
| Popis: | Efficient succinate production in Escherichia coli is attained during anaerobic glucose fermentation in biosynthetic processes combining the reductive branch of the TCA cycle and the glyoxylate bypass. Pyruvate dehydrogenase (PDH) or pyruvate formate lyase (PFL) serves in E. coli as a source of acetyl-CoA, a substrate for the glyoxylate bypass. Depending on enzymes responsible for acetyl-CoA generation, the contribution of the glyoxylate bypass to the anaerobic succinate biosynthesis may vary to support redox balance resulting in diverse maximum achievable yield values. Anaerobic succinate biosynthesis from glucose was studied using E. coli strains with altered expression of genes encoding PFL and PDH. For acetyl-CoA formation by PFL, the yield of 1.32 mol succinate per mole of glucose was achieved with the theoretical value of 1.6 mol/mol. Involvement of PDH in anaerobic acetyl-CoA synthesis increased succinate yield up to 1.49 mol/mol, which is 89.8% of the predicted maximum (1.6(6) mol/mol). The maximum yield of 1.69 mol succinate per mol glucose, amounting to 98.8% of the stoichiometric maximum (1.71 mol/mol), was achieved with the strain possessing PDH as the primary anaerobic source of acetyl-CoA. During high cell density fermentation, the best engineered strain produced high amounts of succinate (570.7 mM) and only small quantities of acetate (11.9 mM). |
| Databáze: | OpenAIRE |
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