Journal of Industrial Microbiology & Biotechnology

, Volume 45, Issue 9, pp 781–788 | Cite as

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

  • Yu Zheng
  • Yangang Chang
  • Renkuan Zhang
  • Jia Song
  • Ying Xu
  • Jing Liu
  • Min WangEmail author
Fermentation, Cell Culture and Bioengineering - Original Paper


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.


Acetic acid fermentation Acetobacter pasteurianus Energy metabolism Oxygen supply 



This work was supported by National Natural Science Foundation of China (31471722, 31671851), National Key R&D Program of China (2016YFD0400505), Tianjin Municipal Science and Technology Commission (16YFZCNC00650, 17PTGCCX00190), Rural Affairs Committee of Tianjin (201701180) and the Innovative Research Team of Tianjin Municipal Education Commission (TD13-5013).

Author Contributions

YZ and YC conceived and designed the study. RZ, YX and JL performed the experiments. JS reviewed and edited the manuscript. MW is the PI who received the grant and coordinated the project. All authors have read and approved the manuscript.

Supplementary material

10295_2018_2060_MOESM1_ESM.docx (15 kb)
Supplementary material 1 (DOCX 15 kb)


  1. 1.
    Adler P, Frey LJ, Berger A, Bolten CJ, Hansen CE, Wittmann C (2014) The key to acetate: metabolic fluxes of acetic acid bacteria under cocoa pulp fermentation-simulating conditions. Appl Environ Microbiol 80:4702–4716. CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Andrésbarrao C, Saad MM, Chappuis ML, Boffa M, Perret X, Ortega PR, Barja F (2012) Proteome analysis of Acetobacter pasteurianus during acetic acid fermentation. J Proteom 75:1701–1717. CrossRefGoogle Scholar
  3. 3.
    Bennett GN, San KY (2017) Strategies for manipulation of oxygen utilization by the electron transfer chain in microbes for metabolic engineering purposes. J Ind Microbiol Biotechnol 44:647–658. CrossRefPubMedGoogle Scholar
  4. 4.
    Fukaya M, Takemura H, Okumura H, Kawamura Y, Horinouchi S, Beppu T (1990) Cloning of genes responsible for acetic acid resistance in Acetobacter aceti. J Bacteriol 172:2096–2104. CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Fukaya M, Takemura H, Tayama K, Okumura H, Kawamura Y, Horinouchi S, Beppu T (1993) The aarC gene responsible for acetic acid assimilation confers acetic acid resistance on Acetobacter aceti. J Ferment Bioeng 76:270–275. CrossRefGoogle Scholar
  6. 6.
    Gullo M, Verzelloni E, Canonico M (2014) Aerobic submerged fermentation by acetic acid bacteria for vinegar production: process and biotechnological aspects. Process Biochem 49:1571–1579. CrossRefGoogle Scholar
  7. 7.
    Gullo M, Zanichelli G, Verzelloni E, Lemmetti F, Giudici P (2016) Feasible acetic acid fermentations of ethanolic and sugary substrates in combined operation mode. Process Biochem 51:1129–1139. CrossRefGoogle Scholar
  8. 8.
    Hitschmann A, Stockinger H (1985) Oxygen deficiency and its effect on the adenylate system in Acetobacter in the submerse acetic fermentation. Appl Microbiol Biot 22:46–49. CrossRefGoogle Scholar
  9. 9.
    Ivanina AV, Sokolov EP, Sokolova IM (2010) Effects of cadmium on anaerobic energy metabolism and mRNA expression during air exposure and recovery of an intertidal mollusk Crassostrea virginica. Aquat Toxicol 99:330–342CrossRefPubMedGoogle Scholar
  10. 10.
    Mamlouk D, Gullo M (2013) Acetic acid bacteria: physiology and carbon sources oxidation. Indian J Microbiol 53:377–384. CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Matsushita K, Inoue T, Adachi O, Toyama H (2005) Acetobacter aceti possesses a proton motive force-dependent efflux system for acetic acid. J Bacteriol 187:4346–4352. CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Mullins EA, Francois JA, Kappock TJ (2008) A specialized citric acid cycle requiring succinyl-coenzyme A (CoA): acetate CoA-transferase (AarC) confers acetic acid resistance on the acidophile Acetobacter aceti. J Bacteriol 190:4933–4940. CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Muraoka H, Ogasawara N, Takahashi H (1983) Trigger of damage by oxygen deficiency to the acid production system during submerged acetic fermentation with Acetobacter aceti. J Ferment Technol 61:89–93Google Scholar
  14. 14.
    Nakano S, Fukaya M (2008) Analysis of proteins responsive to acetic acid in Acetobacter: molecular mechanisms conferring acetic acid resistance in acetic acid bacteria. Int J Food Microbiol 125:54–59. CrossRefPubMedGoogle Scholar
  15. 15.
    Okamoto-Kainuma A, Yan W, Fukaya M, Tukamoto Y, Ishikawa M, Koizumi Y (2004) Cloning and characterization of the dnaKJ operon in Acetobacter aceti. J Biosci Bioeng 97:339–342. CrossRefPubMedGoogle Scholar
  16. 16.
    Qi ZL, Yang HL, Xia XL, Xin Y, Zhang L, Wang W, Yu XB (2013) A protocol for optimization vinegar fermentation according to the ratio of oxygen consumption versus acid yield. J Food Eng 116:304–309. CrossRefGoogle Scholar
  17. 17.
    Qi ZL, Yang HL, Xia XL, Quan W, Wang W, Yu XB (2014) Achieving high strength vinegar fermentation via regulating cellular growth status and aeration strategy. Process Biochem 49:1063–1070. CrossRefGoogle Scholar
  18. 18.
    Qi ZL, Yang HL, Xia XL, Wang W, Yu XB (2014) High strength vinegar fermentation by Acetobacter pasteurianus via enhancing ethanol respiratory chain. Biotechnol Bioprocess E 19:289–297. CrossRefGoogle Scholar
  19. 19.
    Saichana N, Matsushita K, Adachi O, Frebort I, Frebortova J (2015) Acetic acid bacteria: a group of bacteria with versatile biotechnological applications. Biotechnol Adv 33:1260–1271. CrossRefPubMedGoogle Scholar
  20. 20.
    Sakurai K, Yamazaki S, Ishii M, Igarashi Y, Arai H (2013) Role of the glyoxylate pathway in acetic acid production by Acetobacter aceti. J Biosci Bioeng 115:32–36. CrossRefPubMedGoogle Scholar
  21. 21.
    Solieri L, Giudici P (2009) Vinegars of the world. Springer, Milan (pp1–16) CrossRefGoogle Scholar
  22. 22.
    Trcek J, Toyama H, Czuba J, Misiewicz A, Matsushita K (2006) Correlation between acetic acid resistance and characteristics of PQQ-dependent ADH in acetic acid bacteria. Appl Microbiol Biotechnol 70:366–373. CrossRefPubMedGoogle Scholar
  23. 23.
    Trcek J, Mira NP, Jarboe LR (2015) Adaptation and tolerance of bacteria against acetic acid. Appl Microbiol Biotechnol 99:6215–6229. CrossRefPubMedGoogle Scholar
  24. 24.
    Wang B, Shao YC, Chen FS (2015) Overview on mechanisms of acetic acid resistance in acetic acid bacteria. World J Microbiol Biotechnol 31:255–263. CrossRefPubMedGoogle Scholar
  25. 25.
    Wang B, Shao YC, Chen T, Chen WP, Chen FS (2015) Global insights into acetic acid resistance mechanisms and genetic stability of Acetobacter pasteurianus strains by comparative genomics. Sci Rep 5:18330. CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Zheng Y, Zhang RK, Yin HS, Bai XL, Chang YG, Xia ML, Wang M (2017) Acetobacter pasteurianus metabolic change induced by initial acetic acid to adapt to acetic acid fermentation conditions. Appl Microbiol Biotechnol 101:7007–7016. CrossRefPubMedGoogle Scholar
  27. 27.
    Zheng Y, Chang YG, Xie SK, Song J, Wang M (2018) Impacts of bioprocess engineering on product formation by Acetobacter pasteurianus. Appl Microbiol Biotechnol 102:2535–2541. CrossRefPubMedGoogle Scholar

Copyright information

© Society for Industrial Microbiology and Biotechnology 2018

Authors and Affiliations

  • Yu Zheng
    • 1
  • Yangang Chang
    • 1
  • Renkuan Zhang
    • 1
  • Jia Song
    • 1
  • Ying Xu
    • 1
  • Jing Liu
    • 1
  • Min Wang
    • 1
    Email author
  1. 1.State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Engineering Research Center of Microbial Metabolism and Fermentation Process Control, College of BiotechnologyTianjin University of Science and TechnologyTianjinPeople’s Republic of China

Personalised recommendations