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Applied Microbiology and Biotechnology

, Volume 85, Issue 3, pp 741–751 | Cite as

Characterization of thermotolerant Acetobacter pasteurianus strains and their quinoprotein alcohol dehydrogenases

  • Watchara Kanchanarach
  • Gunjana Theeragool
  • Toshiharu Yakushi
  • Hirohide Toyama
  • Osao Adachi
  • Kazunobu Matsushita
Applied Microbial and Cell Physiology

Abstract

We isolated several thermotolerant Acetobacter species of which MSU10 strain, identified as Acetobacter pasteurianus, could grow well on agar plates at 41°C, tolerate to 1.5% acetic acid or 4% ethanol at 39°C, similarly seen with A. pasteurianus SKU1108 previously isolated. The MSU10 strain showed higher acetic acid productivity in a medium containing 6% ethanol at 37°C than SKU1108 while SKU1108 strain could accumulate more acetic acid in a medium supplemented with 4–5% ethanol at the same temperature. The fermentation ability at 37°C of these thermotolerant strains was superior to that of mesophilic A. pasteurianus IFO3191 strain having weak growth and very delayed acetic acid production at 37°C even at 4% ethanol. Alcohol dehydrogenases (ADHs) were purified from MSU10, SKU1108, and IFO3191 strains, and their properties were compared related to the thermotolerance. ADH of the thermotolerant strains had a little higher optimal temperature and heat stability than that of mesophilic IFO3191. More critically, ADHs from MSU10 and SKU1108 strains exhibited a higher resistance to ethanol and acetic acid than IFO3191 enzyme at elevated temperature. Furthermore, in this study, the ADH genes were cloned, and the amino acid sequences of ADH subunit I, subunit II, and subunit III were compared. The difference in the amino acid residues could be seen, seemingly related to the thermotolerance, between MSU10 or SKU1108 ADH and IFO 3191 ADH.

Keywords

Acetic acid bacteria Acetobacter pasteurianus Quinoprotein alcohol dehydrogenases Acetic acid fermentation 

Notes

Acknowledgments

This work was supported by the Program for Promotion of Basic Research Activities for Innovative Biosciences. Part of this work was carried out through collaboration in a Core University Program between Yamaguchi University and Kasetsart University supported by The Japan Society for the Promotion of Science (JSPS) and the National Research Council of Thailand (NRCT). W. K. would like to thank Mahasarakham University for the financial support to pursue her Ph.D. program. We are grateful to Dr. Mamoru Yamada for his helpful discussion during the course of this work.

Supplementary material

253_2009_2203_MOESM1_ESM.doc (64 kb)
Table S1 Growth characteristics of acetic acid bacteria (22 isolated strains; DOC 63.5 kb)
253_2009_2203_MOESM2_ESM.doc (35 kb)
Table S2 Utilization of carbon sources (DOC 35 kb)
253_2009_2203_MOESM3_ESM.doc (40 kb)
Table S3 Comparison of 16S rRNA sequences of Acetobacter sp. MSU10 and MSU22 with those of other acetic acid bacteria (DOC 40 kb)
253_2009_2203_MOESM4_ESM.doc (38 kb)
Table S4 Purification summary of ADHs from A. pasteurianus strains (DOC 38 kb)
253_2009_2203_MOESM5_ESM.doc (40 kb)
Table S5 Substrate specificity of ADHs from A. pasteurianus strains (DOC 39.5 kb)
253_2009_2203_MOESM6_ESM.doc (48 kb)
Fig. S1 An alignment of amino acid sequence of ADH subunit I (a), subunit II (b), and subunit III (c) from A. pasteurianus MSU10, SKU1108, and IFO3191. The black bar and white letters indicate different amino acid residues. W-motifs (W1–W8) are indicated as black boxes. Amino acid residues involved in PQQ binding are bold letters, and those involved in heme c binding are gray areas (Toyama et al. 2004; DOC 48 kb)

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Watchara Kanchanarach
    • 1
  • Gunjana Theeragool
    • 2
  • Toshiharu Yakushi
    • 1
  • Hirohide Toyama
    • 3
  • Osao Adachi
    • 1
  • Kazunobu Matsushita
    • 1
  1. 1.Department of Biological Chemistry, Faculty of AgricultureYamaguchi UniversityYamaguchiJapan
  2. 2.Department of Microbiology, Faculty of ScienceKasetsart UniversityBangkokThailand
  3. 3.Department of Bioscience and Biotechnology, Faculty of AgricultureUniversity of the RyukyusOkinawaJapan

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