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. 2024 Jan 9:51:kuae036.
doi: 10.1093/jimb/kuae036.

Improving the alcohol respiratory chain and energy metabolism by enhancing PQQ synthesis in Acetobacter pasteurianus

Wenqing Zhang  1 , Chen Feng  1 , Chunxue Zhang  1 , Jia Song  1 , Li Li  2 , Menglei Xia  1 , Wei Ding  3 , Yu Zheng  1   3   4 , Min Wang  1
Affiliations

Improving the alcohol respiratory chain and energy metabolism by enhancing PQQ synthesis in Acetobacter pasteurianus

Wenqing Zhang et al. J Ind Microbiol Biotechnol. .

Abstract

Pyrroloquinoline quinone (PQQ) is one of the important coenzymes in living organisms. In acetic acid bacteria (AAB), it plays a crucial role in the alcohol respiratory chain, as a coenzyme of alcohol dehydrogenase (ADH). In this work, the PQQ biosynthetic genes were overexpressed in Acetobacter pasteurianus CGMCC 3089 to improve the fermentation performance. The result shows that the intracellular and extracellular PQQ contents in the recombinant strain A. pasteurianus (pBBR1-p264-pqq) were 152.53% and 141.08% higher than those of the control A. pasteurianus (pBBR1-p264), respectively. The catalytic activity of ADH and aldehyde dehydrogenase increased by 52.92% and 67.04%, respectively. The results indicated that the energy charge and intracellular ATP were also improved in the recombinant strain. The acetic acid fermentation was carried out using a 5 L self-aspirating fermenter, and the acetic acid production rate of the recombinant strain was 23.20% higher compared with the control. Furthermore, the relationship between the PQQ and acetic acid tolerance of cells was analyzed. The biomass of recombinant strain was 180.2%, 44.3%, and 38.6% higher than those of control under 2%, 3%, and 4% acetic acid stress, respectively. After being treated with 6% acetic acid for 40 min, the survival rate of the recombinant strain was increased by 76.20% compared with the control. Those results demonstrated that overexpression of PQQ biosynthetic genes increased the content of PQQ, therefore improving the acetic acid fermentation and the cell tolerance against acetic acid by improving the alcohol respiratory chain and energy metabolism.

One sentence summary: The increase in PQQ content enhances the activity of the alcohol respiratory chain of Acetobacter pasteurianus, and the increase in energy charge enhances the tolerance of cells against acetic acid, therefore, improving the efficiency of acetic acid fermentation.

Keywords: Acetobacter pasteurianus; Acetic acid fermentation; Acetic acid tolerance; Energy charge; pyrroloquinoline quinone.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Graphical Abstract
Graphical Abstract
Fig. 1.
Fig. 1.
Effects of overexpressing pqq gene clusters on pyrroloquinoline quinone (PQQ) formation and enzymes catalytic activity. (A) The intracellular and extracellular PQQ contents. (B) Catalytic activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). Harvesting cells during the logarithmic growth phase to detect ADH and ALDH enzyme activity.
Fig. 2.
Fig. 2.
Effect of enhanced pyrroloquinoline quinone (PQQ) formation on acetic acid fermentation. (A) Time curves of acetic acid production and cell growth. (B) Time curves of catalytic activities of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). (C) Time curves of ethanol and glucose consumption. (D) The specific production rate of acetic acid. (E) Time curves of ATP and energy charge (EC).
Fig. 3.
Fig. 3.
The effect of enhanced pyrroloquinoline quinone (PQQ) on acetic acid tolerance. (A) Cell growth. Cultivate the strain in GY medium with initial acetic acid concentrations of 0%, 1%, 2%, 3%, and 4% (v/v) for 48 hr. (B) Cell survival rate. The cells were treated with GYA media containing 2%, 4%, and 6% of acetic acid for 40 min.
Fig. 4.
Fig. 4.
The regulation of genes transcription by overexpressing pyrroloquinoline quinone (PQQ). (A) Transcription levels of genes related to acetic acid tolerance and energy metabolism. The cells were collected when the OD reached about 0.6; (B) Schematic diagram of metabolic network.
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