Impacts of bioprocess engineering on product formation by Acetobacter pasteurianus.

Aerobic Acetobacter pasteurianus is one of the most widely used bacterial species for acetic acid and vinegar production. The acetic acid condition is the primary challenge to the industrial application of A. pasteurianus. Thus, numerous endeavors, including strain improvement and process control, have been performed to improve the product formation and acetic acid tolerance of A. pasteurianus. The metabolic features of A. pasteurianus have been gradually elucidated through omic techniques, such as genomics and proteomics. In this mini review, we summarized bioprocess engineering methods that improved product formation of A. pasteurianus by exploiting its metabolic features. Moreover, given that A. pasteurianus is an important functional microorganism in traditional vinegar production, we discuss its metabolism when cocultured with other microorganisms in traditional vinegar production.

Improving the acetic acid tolerance and fermentation of Acetobacter pasteurianus by nucleotide excision repair protein UvrA.

Acetic acid bacteria (AAB) are widely used in acetic acid fermentation due to their remarkable ability to oxidize ethanol and high tolerance against acetic acid. In Acetobacter pasteurianus, nucleotide excision repair protein UvrA was up-regulated 2.1 times by acetic acid when compared with that without acetic acid. To study the effects of UvrA on A. pasteurianus acetic acid tolerance, uvrA knockout strain AC2005-ΔuvrA, uvrA overexpression strain AC2005 (pMV24-uvrA), and the control strain AC2005 (pMV24), were constructed. One percent initial acetic acid was almost lethal to AC2005-ΔuvrA. However, the biomass of the UvrA overexpression strain was higher than that of the control under acetic acid concentrations. After 6% acetic acid shock for 20 and 40 min, the survival ratios of AC2005 (pMV24-uvrA) were 2 and 0.12%, respectively; however, they were 1.5 and 0.06% for the control strain AC2005 (pMV24). UvrA overexpression enhanced the acetification rate by 21.7% when compared with the control. The enzymes involved in ethanol oxidation and acetic acid tolerance were up-regulated during acetic acid fermentation due to the overexpression of UvrA. Therefore, in A. pasteurianus, UvrA could be induced by acetic acid and is related with the acetic acid tolerance by protecting the genome against acetic acid to ensure the protein expression and metabolism.

Two-stage oxygen supply strategy based on energy metabolism analysis for improving acetic acid production by Acetobacter pasteurianus.

Oxygen acts as the electron acceptor to oxidize ethanol by acetic acid bacteria during acetic acid fermentation. In this study, the energy release rate from ethanol and glucose under different aerate rate were compared, and the relationship between energy metabolism and acetic acid fermentation was analyzed. The results imply that proper oxygen supply can maintain the reasonable energy metabolism and cell tolerance to improve the acetic acid fermentation. Further, the transcriptions of genes that involve in the ethanol oxidation, TCA cycle, ATP synthesis and tolerance protein expression were analyzed to outline the effect of oxygen supply on cell metabolism of Acetobacter pasteurianus. Under the direction of energy metabolism framework a rational two-stage oxygen supply strategy was established to release the power consumption and substrates volatilization during acetic acid fermentation. As a result, the acetic acid production rate of 1.86 g/L/h was obtained, which were 20.78% higher than that of 0.1 vvm one-stage aerate rate. And the final acetic acid concentration and the stoichiometric yield were 88.5 g/L and 94.1%, respectively, which were 84.6 g/L and 89.5% for 0.15 vvm one-stage aerate rate.

Acetobacter pasteurianus metabolic change induced by initial acetic acid to adapt to acetic acid fermentation conditions.

Initial acetic acid can improve the ethanol oxidation rate of acetic acid bacteria for acetic acid fermentation. In this work, Acetobacter pasteurianus was cultured in ethanol-free medium, and energy production was found to increase by 150% through glucose consumption induced by initial acetic acid. However, oxidation of ethanol, instead of glucose, became the main energy production pathway when upon culturing ethanol containing medium. Proteome assay was used to analyze the metabolism change induced by initial acetic acid, which provided insight into carbon metabolic and energy regulation of A. pasteurianus to adapt to acetic acid fermentation conditions. Results were further confirmed by quantitative real-time PCR. In summary, decreased intracellular ATP as a result of initial acetic acid inhibition improved the energy metabolism to produce more energy and thus adapt to the acetic acid fermentation conditions. A. pasteurianus upregulated the expression of enzymes related to TCA and ethanol oxidation to improve the energy metabolism pathway upon the addition of initial acetic acid. However, enzymes involved in the pentose phosphate pathway, the main pathway of glucose metabolism, were downregulated to induce a change in carbon metabolism. Additionally, the enhancement of alcohol dehydrogenase expression promoted ethanol oxidation and strengthened the acetification rate, thereby producing a strong proton motive force that was necessary for energy production and cell tolerance to acetic acid.