CoChat
Sign Up
Cited by 32Microbial Metabolic Engineering and BioproductionMicrobial metabolism and enzyme functionBiochemical and biochemical processes

The DOI matches its Crossref record.

Improving Acetic Acid Production by Over-Expressing PQQ-ADH in Acetobacter pasteurianus

Open access
Wu, Xuefeng, Yao, Hongli, Cao, Lili, Zheng, Zhi, Chen, Xiaoju, Zhang, Min, Wei, Zhaojun, Cheng, Jieshun, Jiang, Shaotong, Pan, Lijun, Li, Xingjiang·2017·Frontiers in Microbiology
DOI 10.3389/fmicb.2017.01713PMC PMC5592214
Open this paper in CoChat — free accountOpen PDF

Abstract

Pyrroquinoline quinone-dependent alcohol dehydrogenase (PQQ-ADH) is a key enzyme in the ethanol oxidase respiratory chain of acetic acid bacteria. To investigate the effect of PQQ-ADH on acetic acid production by Acetobacter pasteurianus JST-S, subunits I (adhA) and II (adhB) of PQQ-ADH were over-expressed, the fermentation parameters and the metabolic flux analysis were compared in the engineered strain and the original one. The acetic acid production was improved by the engineered strain (61.42 g L-1) while the residual ethanol content (4.18 g L-1) was decreased. Analysis of 2D maps indicated that 19 proteins were differently expressed between the two strains; of these, 17 were identified and analyzed by mass spectrometry and two-dimensional gel electrophoresis. With further investigation of metabolic flux analysis of the pathway from ethanol and glucose, the results reveal that over-expression of PQQ-ADH is an effective way to improve the ethanol oxidation respiratory chain pathway and these can offer theoretical references for potential mechanism of metabolic regulation in acetic acid bacteria and researches with its acetic acid resistance.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.3389/fmicb.2017.01713

Cites

Statistics

Figures

Figure 1 Construction of recombinant plasmid PBBR-adhA-adhB.
Figure 1 Construction of recombinant plasmid PBBR-adhA-adhB.CC BY · PubMed Central
Figure 2 PCR amplification products shown by agarose gel electrophoresis. Lanes M, marker (bp);1, Padh; 2, adhA; 3, adhB.
Figure 2 PCR amplification products shown by agarose gel electrophoresis. Lanes M, marker (bp);1, Padh; 2, adhA; 3, adhB.CC BY · PubMed Central
Figure 3 Verification of recombinant plasmids by restriction enzyme digeston. Lanes: M, 8-kbp marker, 1, PBBR-adhA digested with Bam HI and Spe I; 2, PBBR-adhB digested with Spe I and Xba I; and 3, PBBR-adhA-adhB digested with Bam H I.
Figure 3 Verification of recombinant plasmids by restriction enzyme digeston. Lanes: M, 8-kbp marker, 1, PBBR-adhA digested with Bam HI and Spe I; 2, PBBR-adhB digested with Spe I and Xba I; and 3, PBBR-adhA-adhB digested with Bam H I.CC BY · PubMed Central
Figure 4 SDS-PAGE of recombinant protiens over-expressed in the genetically engineered strain Lanes: M, molecular weight marker (KDa); 1 and 2, original strain; 3, engineered strain.
Figure 4 SDS-PAGE of recombinant protiens over-expressed in the genetically engineered strain Lanes: M, molecular weight marker (KDa); 1 and 2, original strain; 3, engineered strain.CC BY · PubMed Central
Figure 5 Comparison of fermentation parameters in the original and engineered strains. ADH activity of the original strain(□) and engineered strain(■) were compared under the different content of ethanol in (A). In (B), white triangle and black triangle represent total acid content of the original strain and engineered strain, respectively. Similarly, white circle and black circle represent ethanol content of the original strain and engineered strain, respectively. Error bars show standard deviations of three replicated measurements.
Figure 5 Comparison of fermentation parameters in the original and engineered strains. ADH activity of the original strain(□) and engineered strain(■) were compared under the different content of ethanol in (A). In (B), white triangle and black triangle represent total acid content of the original strain and engineered strain, respectively. Similarly, white circle and black circle represent ethanol content of the original strain and engineered strain, respectively. Error bars show standard deviations of three replicated measurements.CC BY · PubMed Central
Figure 6 Comparison of 2D-PAGE patterns between original engineered strains. (A) Primary 2D-PAGE maps. (B) 2D-PAGE maps analyzed using PD Quest 8.0 software. Protein spots whose expression level changed more than 50% were identified as differentially expressed proteins.
Figure 6 Comparison of 2D-PAGE patterns between original engineered strains. (A) Primary 2D-PAGE maps. (B) 2D-PAGE maps analyzed using PD Quest 8.0 software. Protein spots whose expression level changed more than 50% were identified as differentially expressed proteins.CC BY · PubMed Central
Figure 7 The metabolic network of A. pasteurianus JST-S Glc, glucose; G6P, glucose-6-phhospahte; F6P, fructose-6-phosphate; Ru5P, ribulose 5-posphate; G3P, glyceraldehydes-3-phosphate; PEP, poshonolpyruvate; Pyr, pyruvate; Pyr, pyruvate; AC.H., Acetaldehyde; OAA, oxaloacetate; Acetyl-CoA, acetyl coenzyme A; CoA, Coenzyme A; Lac, Lactate; Ace, Acetate; EtOH, ethanol; Cit, citrate; Isoc, Isocitrate; α-Ket, α-ketoglutaric acid; Suc, succinic acid; Fum, fumarate; Mal, Malate; BM, biomass.
Figure 7 The metabolic network of A. pasteurianus JST-S Glc, glucose; G6P, glucose-6-phhospahte; F6P, fructose-6-phosphate; Ru5P, ribulose 5-posphate; G3P, glyceraldehydes-3-phosphate; PEP, poshonolpyruvate; Pyr, pyruvate; Pyr, pyruvate; AC.H., Acetaldehyde; OAA, oxaloacetate; Acetyl-CoA, acetyl coenzyme A; CoA, Coenzyme A; Lac, Lactate; Ace, Acetate; EtOH, ethanol; Cit, citrate; Isoc, Isocitrate; α-Ket, α-ketoglutaric acid; Suc, succinic acid; Fum, fumarate; Mal, Malate; BM, biomass.CC BY · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

No references could be resolved yet.

Research works printed but not linked: 44

  1. Andrés-Barrao, 2012, Proteome analysis of Acetobacter pasteurianus, during acetic acid fermentation, J. Proteomics, 75, 1701, 10.1016/j.jprot.2011.11.027 · DOI
  2. Antoniewicz, 2015, Methods and advances in metabolic flux analysis: a mini-review, J. Ind. Microbiol. Biotechnol., 42, 317, 10.1007/s10295-015-1585-x · DOI
  3. Chen, 2016, Correlation between ethanol resistance and characteristics of PQQ-dependent ADH in acetic acid bacteria, Eur. Food. Res. Technol., 242, 837, 10.1007/s00217-015-2589-5 · DOI
  4. Chinnawirotpisan, 2003, Quinoprotein alcohol dehydrogenase is involved in catabolic acetate production, while NAD-dependent alcohol dehydrogenase in ethanol assimilation in Acetobacter pasteurianusSKU1108, J. Biosci. Bioeng., 96, 564, 10.1016/S1389-1723(04)70150-4 · DOI
  5. Dandekar, 2014, Software applications toward quantitative metabolic flux analysis and modeling, Brief. Bionform., 15, 91, 10.1093/bib/bbs065 · DOI
  6. Hartl, 2002, Molecular chaperones in the cytosol: from nascent chain to folded protein, Science, 295, 1852, 10.1126/science.1068408 · DOI
  7. Hesterkamp, 1996, Escherichia coli trigger factor is a prolyl isomerase that associates with nascent polypeptide chains, Proc. Natl. Acad. Sci. U.S.A., 93, 4437, 10.1073/pnas.93.9.4437 · DOI
  8. Illeghems, 2013, Complete genome sequence and comparative analysis of Acetobacter pasteurianus 386B, a strain well-adapted to the cocoa bean fermentation ecosystem, BMC Genomics, 14, 526, 10.1186/1471-2164-14-526 · DOI
  9. Kanchanarach, 2010, Characterization of thermotolerant Acetobacter pasteurianus strains and their quinoprotein alcohol dehydrogenases, Appl. Microbiol. Biot., 85, 741, 10.1007/s00253-009-2203-5 · DOI
  10. Kondo, 1995, Cloning, sequencing, and characterization of the gene encoding the smallest subunit of the three-component membrane-bound alcohol dehydrogenase from Acetobacter pasteurianus, J. Bacteriol, 177, 5048, 10.1128/jb.177.17.5048-5055.1995 · DOI
  11. Kovach, 1995, Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes, Gene, 166, 175, 10.1016/0378-1119(95)00584-1 · DOI
  12. Krahulec, 2003, Characterisation of plasmids purified from Acetobacter pasteurianus 2374, Biochem. Biophys. Res. Commun., 310, 94, 10.1016/j.bbrc.2003.08.125 · DOI
  13. Krusong, 2015, Influence of calcium chloride in the high temperature acetification by strain Acetobacter aceti WK for vinegar, J. Appl. Microbiol., 119, 1291, 10.1111/jam.12930 · DOI
  14. Lariviere, 2001, Uniform binding of aminoacyl-tRNAs to elongation factor Tu by thermodynamic compensation, Science, 294, 165, 10.1126/science.1064242 · DOI
  15. Lasko, 1997, Acetate-specific stress response in acetate-resistant bacteria: an analysis of protein patterns, Biotechnol. Progr., 13, 519, 10.1021/bp970075f · DOI
  16. Li, 2014, High levels of malic acid production by the bioconversion of corn straw hydrolyte using an isolated Rhizopus Delemar Strain, Biotechnol. Bioproce. E., 19, 478, 10.1007/s12257-014-0047-z · DOI
  17. Li, 2009, Screening, breeding and metabolic modulating of a strain producing succinic acid with corn straw hydrolyte, World J. Microbiol. Biotechnol., 25, 667, 10.1007/s11274-008-9936-7 · DOI
  18. Longacre, 1997, Flux analysis of glucose metabolism in Rhizopus oryzae for the purpose of increasing lactate yields, Fungal Genet. Biol., 21, 30, 10.1006/fgbi.1996.0952 · DOI
  19. Lu, 2013, L-glutamine provides acid resistance for Escherichia coli through enzymatic release of ammonia, Cell Res., 23, 635, 10.1038/cr.2013.13 · DOI
  20. Matsushita, 2008, A tightly bound quinone functions in the ubiquinone reaction sites of quinoprotein alcohol dehydrogenase of an acetic acid bacterium, Gluconobacter suboxydans, Biosci. Biotech. Bioch., 72, 2723, 10.1271/bbb.80363 · DOI
  21. Matsushita, 1992, Ethanol oxidase respiratory chain of acetic acid bacteria. reactivity with ubiquinone of pyrroloquinoline quinone-dependent alcohol dehydrogenases purified from Acetobacter aceti, Gluconobacter suboxydans. Biosci. Biotech. Bioch., 56, 304, 10.1271/bbb.56.304 · DOI
  22. Nakano, 2008, Analysis of proteins responsive to acetic acid in acetobacter: molecular mechanisms conferring acetic acid resistance in acetic acid bacteria, Int. J. Food Microbio., 125, 54, 10.1016/j.ijfoodmicro.2007.05.015 · DOI
  23. Nakano, 2004, Enhanced expression of aconitase raises acetic acid resistance, Acetobacter aceti. Fems. Microbio. Lett., 235, 315, 10.1111/j.1574-6968.2004.tb09605.x · DOI
  24. Okamoto-Kainuma, 2004, Cloning and characterization of the dnaKJ operon in Acetobacter aceti, J. Biosci. Bioeng., 97, 339, 10.1016/S1389-1723(04)70216-9 · DOI
  25. Okamoto-Kainuma, 2002, Cloning and characterization of groESL operon in Acetobacter aceti, J. Biosci. Bioeng., 94, 140, 10.1016/S1389-1723(02)80134-7 · DOI
  26. Qi, 2014, Mutation of Acetobacter pasteurianus, by UV irradiation under acidic stress for high-acidity vinegar fermentation, Int. J. Food Sci. Techno., 49, 468, 10.1111/ijfs.12324 · DOI
  27. Qin, 2016, Microbial diversity biochemical analysis of Suanzhou: a traditional Chinese fermented cereal gruel, Front. Microbiol., 7, 1311, 10.3389/fmicb.2016.01311 · DOI
  28. Quintero, 2009, Quantification of the expression of reference and alcohol dehydrogenase genes of some acetic acid bacteria in different growth condition, J. Appl. Microbio., 106, 666, 10.1111/j.1365-2672.2008.04046.x · DOI
  29. Rhee, 1976, Mechanistic studies of glutamine synthetase from Escherichia coli: kinetic evidence for two reaction intermediates in biosynthetic reaction, Proc. Natl. Acad. Sci. U.S.A., 73, 476, 10.1073/pnas.73.2.476 · DOI
  30. Richier, 2006, Oxidative stress and apoptotic events during thermal stress in the symbiotic sea anemone, anemonia viridis, FEBS J., 273, 4186, 10.1111/j.1742-4658.2006.05414.x · DOI
  31. Ryter, 2007, Mechanisms of cell death in oxidative stress, Antioxid. Redox Sign., 9, 49, 10.1089/ars.2007.9.49 · DOI
  32. Steiner, 2001, Proteins induced during adaptation of Acetobacter aceti to high acetate concentrations, Appl. Environ. Microb., 67, 5474, 10.1128/AEM.67.12.5474-5481.2001 · DOI
  33. Stephanopoulos, 1999, Metabolic fluxes and metabolic engineering, Metab. Eng., 1, 1, 10.1006/mben.1998.0101 · DOI
  34. Toyama, 2004, Quinohemoprotein alcohol dehydrogenases: structure, function, and physiology, Arch. Bioch. Biophys., 428, 10, 10.1016/j.abb.2004.03.037 · DOI · Paper page
  35. Trček, 2013, A unique enzyme of acetic acid bacteria, PQQ-dependent alcohol dehydrogenase, is also present in frateuria aurantia, Appl. Microbiol. Biot., 97, 7369, 10.1007/s00253-013-5007-6 · DOI
  36. Trček, 2015, Adaptation and tolerance of bacteria against acetic acid, Appl. Microbiol. Biot., 99, 6215, 10.1007/s00253-015-6762-3 · DOI
  37. Trček, 2006, Correlation between acetic acid resistance and characteristics of PQQ-dependent ADH in acetic acid bacteria, Appl. Microbio. Biot., 70, 366, 10.1007/s00253-005-0073-z · DOI
  38. Wallenius, 2016, Carbon 13-metabolic flux analysis derived constraint-based metabolic modelling of Clostridium acetobutylicum in stressed chemostat conditions, Bioresour. Technol., 219, 378, 10.1016/j.biortech.2016.07.137 · DOI
  39. Wang, 2015, Comparative proteome of Acetobacter pasteurianus Ab3 during the high acidity rice vinegar fermentation, Appl. Biochem. Biotech., 177, 1573, 10.1007/s12010-015-1838-1 · DOI
  40. Welch, 1993, Heat shock proteins functioning as molecular chaperones: their roles in normal and stressed cells, 71, 10.1007/978-94-011-2108-8_9 · DOI
  41. Yakushi, 2010, Alcohol dehydrogenase of acetic acid bacteria: structure, mode of action, and applications in biotechnology, Appl. Microbiol. Biot., 86, 1257, 10.1007/s00253-010-2529-z · DOI
  42. Yanisch-Perron, 1985, Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors, Gene, 33, 103, 10.1016/0378-1119(85)90120-9 · DOI
  43. Zhang, 2017, Valorization of Spent Escherichia coli media using green microalgae Chlamydomonas reinhardtii and feedstock production, Front. Microbiol., 8, 1026, 10.3389/fmicb.2017.01026 · DOI
  44. Zheng, 2014, Genetic Engineering Acetic Acid Bacteria of Overexpressing Coenzyme PQQ (Pyrroloquinoline Quinone) Synthetic Proteins and Application of Bacteria.

Cited by

See all 32

Metadata

Authors
Wu, X., Yao, H., Cao, L., Zheng, Z., Chen, X., Zhang, M., Wei, Z., Cheng, J., Jiang, S., Pan, L., & Li, X.
Affiliations
  • Hefei University of Technology (CN)
  • Chaohu University (CN)
Corresponding author
Li, Xingjiang
ORCID
Venue
Frontiers in Microbiology
Publisher
Frontiers Media SA
Published
2017-09-06
Volume
8
Pages
1713-1713
ISSN
1664-302X (ISSN-L)
DOI
10.3389/fmicb.2017.01713
PMC
PMC5592214
OpenAlex
W2748870868
Cited by
32
References
0
Citations by year
2026: 1 · 2025: 2 · 2024: 6 · 2023: 5 · 2022: 2 · 2021: 4 · 2020: 9 · 2019: 2 · 2018: 1
FWCI
1.34
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Frontiers in Microbiology (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • National Natural Science Foundation of China (31601465)
  • Hefei University
  • Hefei University of Technology
  • Natural Science Foundation of Anhui Province (1408085MKL17)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial metabolism and enzyme function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Biochemical and biochemical processes
Keywords
acetic acid production, Acetobacter pasteurianus, ethanol oxidation, electron transport chain, metabolic flux analysis, protein expression profiling, acetic acid resistance

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review

How to cite

Cite CoChat for the checks on this page, and the paper for its own findings.

This page
CoChat. (2026). Citation check: Improving Acetic Acid Production by Over-Expressing PQQ-ADH in Acetobacter pasteurianus. https://app.cochat.ai/papers/10.3389/fmicb.2017.01713
The paper
Wu, X., Yao, H., Cao, L., Zheng, Z., Chen, X., Zhang, M., Wei, Z., Cheng, J., Jiang, S., Pan, L., & Li, X. (2017). Improving Acetic Acid Production by Over-Expressing PQQ-ADH in Acetobacter pasteurianus. Frontiers in Microbiology. https://doi.org/10.3389/fmicb.2017.01713