D-Lactic Acid → Lactate

Fermentation of glucose to D-lactic acid under aerobic growth condition by an evolved E. coli mutant deficient in three terminal oxidases is reported in this work. Cytochrome oxidases (cydAB, cyoABCD, cbdAB) were removed from the E. coli K12 MG1655 genome resulting in the ECOM3 (E. coli Cytochrome Oxidase Mutant) strain. Removal of cytochrome oxidases reduced the oxygen uptake rate of the knock-out strain by nearly 85%. Moreover, the knock-out strain was initially incapable of growing on M9 minimal media. After subjecting the ECOM3 strain to adaptive evolution on glucose M9 medium for 60 days, the growth rate equivalent to anaerobic wild type E. coli was achieved. Our findings demonstrate that three independently adaptively evolved ECOM3 populations acquired different phenotypes: one produced lactate as a sole fermentation product while the other two strains exhibited a mixed acid fermentation under oxic growth conditions with lactate remaining as the major product. The homofermenting strain showed the D-lactate yield of 0.8 g/g from glucose. Gene expression and in silico model-based analysis was employed to identify perturbed pathways and explain phenotypic behavior. Significant upregulation of ygiN and sodAB explain the remaining oxygen uptake that was observed in evolved ECOM3 strains. E. coli strains exemplified herein showed the ability to produce lactate as a fermentation product from glucose as well as undergo mixed-acid fermentation during aerobic (oxic conditions) growth.

1. customCytochrome oxidases (cydAB, cyoABCD, cbdAB) were removed from the E
2. customgenome resulting in the ECOM3 (E. coli Cytochrome Oxidase Mutant) strain
3. customRemoval of cytochrome oxidases
4. customfor 60 days

ORD record

ord-2d51e1054e1e486dbf9225e460d20ed8

Source dataset

uspto-grants-2015_07

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US09080178B2