Applied Microbiology and Biotechnology

, Volume 102, Issue 10, pp 4549–4561 | Cite as

Role of a membrane-bound aldehyde dehydrogenase complex AldFGH in acetic acid fermentation with Acetobacter pasteurianus SKU1108

  • Toshiharu Yakushi
  • Seiya Fukunari
  • Tomohiro Kodama
  • Minenosuke Matsutani
  • Shun Nina
  • Naoya Kataoka
  • Gunjana Theeragool
  • Kazunobu Matsushita
Applied microbial and cell physiology


Acetic acid fermentation is widely considered a consequence of ethanol oxidation by two membrane-bound enzymes—alcohol dehydrogenase and aldehyde dehydrogenase (ALDH)—of acetic acid bacteria. Here, we used a markerless gene disruption method to construct a mutant of the Acetobacter pasteurianus strain SKU1108 with a deletion in the aldH gene, which encodes the large catalytic subunit of a heterotrimeric ALDH complex (AldFGH), to examine the role of AldFGH in acetic acid fermentation. The ΔaldH strain grew less on ethanol-containing medium, i.e., acetic acid fermentation conditions, than the wild-type strain and significantly accumulated acetaldehyde in the culture medium. Unexpectedly, acetaldehyde oxidase activity levels of the intact ΔaldH cells and the ΔaldH cell membranes were similar to those of the wild-type strain, which might be attributed to an additional ALDH isozyme (AldSLC). The apparent KM values of the wild-type and ΔaldH membranes for acetaldehyde were similar to each other, when the cells were cultured in nonfermentation conditions, where ΔaldH cells grow as well as the wild-type cells. However, the membranes of the wild-type cells grown under fermentation conditions showed a 10-fold lower apparent KM value than those of the cells grown under nonfermentation conditions. Under fermentation conditions, transcriptional levels of a gene for AldSLC were 10-fold lower than those under nonfermentation conditions, whereas aldH transcript levels were not dramatically changed under the two conditions. We suggest that A. pasteurianus SKU1108 has two ALDHs, and the AldFGH complex is indispensable for acetic acid fermentation and is the major enzyme under fermentation conditions.


Acetic acid fermentation Acetaldehyde Membrane-bound aldehyde dehydrogenase Acetobacter pasteurianus Molybdenum molybdopterin 



We would like to thank Luis Ielpi (Universidad de Buenos Aires, Argentina) for kindly providing pK18mobGII and Megumi Ichiki (Yamaguchi University) for her technical assistance. We are grateful to Osao Adachi (Yamaguchi University) and Hirohide Toyama (University of Ryukyus) who encouraged us throughout this study.


This study was funded by MEXT KAKENHI (grant number 23580115). Part of this work was funded by the Core to Core Program, which was supported by the Japan Society for the Promotion of Science (JSPS) and the National Research Council of Thailand (NRCT).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2018_8940_MOESM1_ESM.pdf (201 kb)
ESM 1 (PDF 200 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Graduate School of Science and Technology for InnovationYamaguchi UniversityYamaguchiJapan
  2. 2.Department of Biological Chemistry, Faculty of AgricultureYamaguchi UniversityYamaguchiJapan
  3. 3.Research Center for Thermotolerant Microbial ResourcesYamaguchi UniversityYamaguchiJapan
  4. 4.Department of Microbiology, Faculty of ScienceKasetsart UniversityBangkokThailand

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