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Introduction: Weak aerobic stability is a notable challenge for whole-plant corn silage, particularly in hot and humid regions. Acetobacter is commonly regarded as an indicator of aerobic deterioration in silage, yet its precise role in fermentation and during aerobic exposure, as well as the factors that promote its growth, remain insufficiently understood. Methods: In this study, whole-plant corn silage was prepared using a bagged method with controlled dry matter (DM) content at 20%, 25%, and 30%, and initial concentrations of A. pasteurianus at 40%, 50%, and 60%. The silage was stored for 60 days under varying temperatures (20°C, 30°C, and 40°C). Following the anaerobic storage phase, the silage was exposed to air at room temperature (20-25°C) for 7 days, both with and without A. pasteurianus inoculation. Results: The results demonstrated that A. pasteurianus did not impact the nutritional value of the silage during anaerobic fermentation, maintaining a low pH (< 3.80). However, during aerobic exposure, the presence of A. pasteurianus significantly reduced the aerobic stability of the silage. The microbial community shifted from primarily Klebsiella species initially to Lactobacillus and Acetobacter species post-ensiling. During the aerobic exposure phase, A. pasteurianus and A. fabarum became the dominant species. Response Surface Methodology (RSM) analysis identified optimal conditions for the proliferation of A. pasteurianus during the aerobic phase, which occurred at 28°C, 25% DM, and 52% initial concentration at 3 ml/kg. Discussion: These findings confirm that A. pasteurianus plays a critical role in reducing the aerobic stability of whole-plant corn silage. Additionally, the study identifies the optimal conditions that favor the proliferation of A. pasteurianus, offering valuable insights for the development of strategies to prevent and control this bacterium, thereby improving the aerobic stability of silage in hot and humid regions. [ABSTRACT FROM AUTHOR]

Lipopolysaccharide (LPS), a cell surface component of Gram‐negative bacteria, activates innate immunity. Its active principle is the terminal glycolipid lipid A. Acetobacter pasteurianus is a Gram‐negative bacterium used in the fermentation of traditional Japanese black rice vinegar (kurozu). In this study, we focused on A. pasteurianus lipid A, which is a potential immunostimulatory component of kurozu. The active principle structure of A. pasteurianus lipid A has not yet been identified. Herein, we first systematically synthesized three types of A. pasteurianus lipid As containing a common and unique tetrasaccharide backbone. We developed an efficient method for constructing the 2‐trehalosamine skeleton utilizing borinic acid‐catalyzed glycosylation to afford 1,1'‐α,α‐glycoside in high yield and stereoselectivity. A common tetrasaccharide intermediate with an orthogonal protecting group pattern was constructed via [2+2] glycosylation. After introducing various fatty acids, all protecting groups were removed to achieve the first chemical synthesis of three distinct types of A. pasteurianus lipid As. After evaluating their immunological function using both human and murine cell lines, we identified the active principles of A. pasteurianus LPS. We also found the unique anomeric structure of A. pasteurianus lipid A contributes to its high chemical stability. [ABSTRACT FROM AUTHOR]

Background: This study explored the utilization of luffa sponge (LS) in enhancing acetification processes. LS is known for having high porosity and specific surface area, and can provide a novel means of supporting the growth of acetic acid bacteria (AAB) to improve biomass yield and acetification rate, and thereby promote more efficient and sustainable vinegar production. Moreover, the promising potential of LS and luffa sponge coated with κ-carrageenan (LSK) means they may represent effective alternatives for the co-production of industrially valuable bioproducts, for example bacterial cellulose (BC) and acetic acid. Methods: LS and LSK were employed as adsorbents for Acetobacter pasteurianus UMCC 2951 in a submerged semi-continuous acetification process. Experiments were conducted under reciprocal shaking at 1 Hz and a temperature of 32 °C. The performance of the two systems (LS-AAB and LSK-AAB respectively) was evaluated based on cell dry weight (CDW), acetification rate, and BC biofilm formation. Results: The use of LS significantly increased the biomass yield during acetification, achieving a CDW of 3.34 mg/L versus the 0.91 mg/L obtained with planktonic cells. Coating LS with κ-carrageenan further enhanced yield, with a CDW of 4.45 mg/L. Acetification rates were also higher in the LSK-AAB system, reaching 3.33 ± 0.05 g/L d as opposed to 2.45 ± 0.05 g/L d for LS-AAB and 1.13 ± 0.05 g/L d for planktonic cells. Additionally, BC biofilm formation during the second operational cycle was more pronounced in the LSK-AAB system (37.0 ± 3.0 mg/L, as opposed to 25.0 ± 2.0 mg/L in LS-AAB). Conclusions: This study demonstrates that LS significantly improves the efficiency of the acetification process, particularly when enhanced with κ-carrageenan. The increased biomass yield, accelerated acetification, and enhanced BC biofilm formation highlight the potential of the LS-AAB system, and especially the LSK-AAB variant, in sustainable and effective vinegar production. These systems offer a promising approach for small-scale, semi-continuous acetification processes that aligns with eco-friendly practices and caters to specialized market needs. Finally, this innovative method facilitates the dual production of acetic acid and bacterial cellulose, with potential applications in biotechnological fields. [ABSTRACT FROM AUTHOR]

Background This study explored the utilization of luffa sponge (LS) in enhancing acetification processes. LS is known for having high porosity and specific surface area, and can provide a novel means of supporting the growth of acetic acid bacteria (AAB) to improve biomass yield and acetification rate, and thereby promote more efficient and sustainable vinegar production. Moreover, the promising potential of LS and luffa sponge coated with κ-carrageenan (LSK) means they may represent effective alternatives for the co-production of industrially valuable bioproducts, for example bacterial cellulose (BC) and acetic acid. Methods LS and LSK were employed as adsorbents for Acetobacter pasteurianus UMCC 2951 in a submerged semi-continuous acetification process. Experiments were conducted under reciprocal shaking at 1 Hz and a temperature of 32 °C. The performance of the two systems (LS-AAB and LSK-AAB respectively) was evaluated based on cell dry weight (CDW), acetification rate, and BC biofilm formation. Results The use of LS significantly increased the biomass yield during acetification, achieving a CDW of 3.34 mg/L versus the 0.91 mg/L obtained with planktonic cells. Coating LS with κ-carrageenan further enhanced yield, with a CDW of 4.45 mg/L. Acetification rates were also higher in the LSK-AAB system, reaching 3.33 ± 0.05 g/L d as opposed to 2.45 ± 0.05 g/L d for LS-AAB and 1.13 ± 0.05 g/L d for planktonic cells. Additionally, BC biofilm formation during the second operational cycle was more pronounced in the LSK-AAB system (37.0 ± 3.0 mg/L, as opposed to 25.0 ± 2.0 mg/L in LS-AAB). Conclusions This study demonstrates that LS significantly improves the efficiency of the acetification process, particularly when enhanced with κ-carrageenan. The increased biomass yield, accelerated acetification, and enhanced BC biofilm formation highlight the potential of the LS-AAB system, and especially the LSK-AAB variant, in sustainable and effective vinegar production. These systems offer a promising approach for small-scale, semi-continuous acetification processes that aligns with eco-friendly practices and caters to specialized market needs. Finally, this innovative method facilitates the dual production of acetic acid and bacterial cellulose, with potential applications in biotechnological fields.

Acetobacter pasteurianus is always used to brew vinegar because of its ability of producing and tolerating a high concentration of acetic acid. During vinegar fermentation, initial acetic acid contributes to acetic acid accumulation, which varies with initial concentrations. In this study, to investigate the mechanisms of tolerating and producing acetic acid of Acetobacter pasteurianus under different concentrations of substrate acetic acid, four-dimensional label-free proteomic technology has been used to analyze the protein profiles of Acetobacter pasteurianus at different growth stages (the lag and exponential phases) and different substrate acetic acid concentrations (0%, 3%, and 6%). A total of 2093 proteins were quantified in this study. The differentially expressed proteins were majorly involved in gene ontology terms of metabolic processes, cellular metabolic processes, and substance binding. Under acetic acid stress, strains might attenuate the toxicity of acetic acid by intensifying fatty acid metabolism, weakening the tricarboxylic acid cycle, glycerophospholipid and energy metabolism during the lag phase, while strains might promote the assimilation of acetic acid and inter-conversion of substances during the exponential phase by enhancing the tricarboxylic acid cycle, glycolysis, pyruvate, and energy metabolism to produce and tolerate acid. Besides, cell cycle regulation and protein translation might be potential acid tolerance pathways under high acid stress. The result contributes to the exploration of new potential acid tolerance mechanisms in Acetobacter pasteurianus from four-dimensional label-free relative quantitative proteomics analysis. [ABSTRACT FROM AUTHOR]

Abstract Quorum‐sensing (QS) is an essential mechanism of microbial self‐regulation. In this work, the presence of QS in Acetobacter pasteurianus was demonstrated, and N‐acyl‐homoserine lactones N‐C4‐HSL and N‐C8‐HSL were identified as the signal molecules. The exogenous addition of N‐C4‐HSL, N‐C8‐HSL, N‐C10‐HSL, and N‐C12‐HSL result in the reduction in biomass, and lead to 11.56%, 11.91%, 7.94%, and 4.69% decrease in acetic acid production, respectively, when compared with the control. A hypothetical N‐acylhomoserine lactonase (AHL lactonase) 0046 was overexpressed in A. pasteurianus CGMCC 3089, and the recombinant strain exhibites a 17.38% and 12.75% increase in the biomass and the acetic acid production rate, respectively. Those results prove the negative regulation of QS on acetic acid fermentation, and overexpression of 0046 protein alleviates this negative effect.

Aims: This study aimed to investigate the probiotic effects of Acetobacter pasteurianus BP2201, isolated from brewing mass, for the treatment of alcohol-induced learning and memory ability impairments in a Caenorhabditis elegans model. Methods and results: Acetobacter pasteurianus BP2201 was examined for probiotic properties, including acid and bile salt resistance, ethanol degradation, antioxidant efficacy, hemolytic activity, and susceptibility to antibiotics. The strain displayed robust acid and bile salt tolerance, efficient ethanol degradation, potent antioxidant activity, and susceptibility to specific antibiotics. Additionally, in the C. elegans model, administering A. pasteurianus BP2201 significantly improved alcohol-induced learning and memory impairments. Conclusions: Acetobacter pasteurianus BP2201 proves to be a promising candidate strain for the treatment of learning and memory impairments induced by alcohol intake. [ABSTRACT FROM AUTHOR]

Acetic acid bacteria have a remarkable capacity to cope with elevated concentrations of cytotoxic acetic acid in their fermentation environment. In particular, the high-level acetate tolerance of Acetobacter pasteurianus that occurs in vinegar industrial settings must be constantly selected for. However, the improved acetic acid tolerance is rapidly lost without a selection pressure. To understand genetic and molecular biology of this acquired acetic acid tolerance in A. pasteurianus, we evolved three strains A. pasteurianus CICIM B7003, CICIM B7003-02, and ATCC 33,445 over 960 generations (4 months) in two initial acetic acids of 20 g·L−1 and 30 g·L−1, respectively. An acetic acid-adapted strain M20 with significantly improved specific growth rate of 0.159 h−1 and acid productivity of 1.61 g·L−1·h−1 was obtained. Comparative genome analysis of six evolved strains revealed that the genetic variations of adaptation were mainly focused on lactate metabolism, membrane proteins, transcriptional regulators, transposases, replication, and repair system. Among of these, lactate dehydrogenase, acetolactate synthase, glycosyltransferase, ABC transporter ATP-binding protein, two-component regulatory systems, the type II toxin-antitoxin system (RelE/RelB/StbE), exodeoxyribonuclease III, type I restriction endonuclease, tRNA-uridine 2-sulfurtransferase, and transposase might collaboratively contribute to the improved acetic acid tolerance in A. pasteurianus strains. The balance between repair factors and transposition variations might be the basis for genomic plasticity of A. pasteurianus strains, allowing the survival of populations and their offspring in acetic acid stress fluctuations. These observations provide important insights into the nature of acquired acetic acid tolerance phenotype and lay a foundation for future genetic manipulation of these strains. [ABSTRACT FROM AUTHOR]

Quorum‐sensing (QS) is an essential mechanism of microbial self‐regulation. In this work, the presence of QS in Acetobacter pasteurianus was demonstrated, and N‐acyl‐homoserine lactones N‐C4‐HSL and N‐C8‐HSL were identified as the signal molecules. The exogenous addition of N‐C4‐HSL, N‐C8‐HSL, N‐C10‐HSL, and N‐C12‐HSL result in the reduction in biomass, and lead to 11.56%, 11.91%, 7.94%, and 4.69% decrease in acetic acid production, respectively, when compared with the control. A hypothetical N‐acylhomoserine lactonase (AHL lactonase) 0046 was overexpressed in A. pasteurianus CGMCC 3089, and the recombinant strain exhibites a 17.38% and 12.75% increase in the biomass and the acetic acid production rate, respectively. Those results prove the negative regulation of QS on acetic acid fermentation, and overexpression of 0046 protein alleviates this negative effect. [ABSTRACT FROM AUTHOR]

Polyphenols are important functional substances produced in the acetic acid fermentation (AAF) of Shanxi aged vinegar (SAV). Previous studies have shown that the metabolic activity of microorganisms is closely related to polyphenol production and accumulation. In this study, microorganisms in the AAF of SAV were analyzed to explore how to increase the polyphenol yield by changing the microorganisms and reveal the potential mechanism of the microbial influence on the polyphenol yield. Macrotranscriptome analysis showed that acetic and lactic acid bacteria dominated the AAF fermentation process and initially increased and decreased. Spearman correlation analysis and verification experiments showed that the co-addition of Acetobacter pasteurianus and Lactobacillus helveticus promoted the accumulation of polyphenols, and the total polyphenol content increased by 72% after strengthening. [ABSTRACT FROM AUTHOR]

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Traditional grain vinegar is fermented using multiple acetic acid bacteria (AAB) at various temperatures. A single AAB showed high acid-producing ability at 30 °C with a 5% alcohol concentration and an initial pH adjusted to 4.0. Multiple AAB were similar to a single AAB; however, the optimal initial pH was 3.0. Acid production ability according to the type of AAB was higher in multiple AAB than in single AAB. That is, using multiple AAB helped increase the titratable acidity of traditional grain vinegar. In addition, increasing the titratable acidity and content of volatile flavor compounds was advantageous when two, rather than four, AAB types were mixed and used. The titratable acidity was high at medium temperatures (30 °C); however, volatile flavor compounds increased at low temperatures (20 °C) under multiple AAB. A 16S rDNA-based microbiome taxonomic profiling analysis identified differences in beta diversity due to multiple AAB and fermentation temperatures. In particular, beta diversity analysis revealed a specific pattern when a mixture of Acetobacter ascedens GV–8 and Acetobacter pasteurianus GV–22 was fermented at a low temperature (20 °C). Therefore, we propose the application of multiple AAB with acidic and flavor-producing properties in traditional grain vinegar. [ABSTRACT FROM AUTHOR]

Acetobacter pasteurianus is always used to brew vinegar because of its ability of producing and tolerating a high concentration of acetic acid. During vinegar fermentation, initial acetic acid contributes to acetic acid accumulation, which varies with initial concentrations. In this study, to investigate the mechanisms of tolerating and producing acetic acid of Acetobacter pasteurianus under different concentrations of substrate acetic acid, four-dimensional label-free proteomic technology has been used to analyze the protein profiles of Acetobacter pasteurianus at different growth stages (the lag and exponential phases) and different substrate acetic acid concentrations (0%, 3%, and 6%). A total of 2093 proteins were quantified in this study. The differentially expressed proteins were majorly involved in gene ontology terms of metabolic processes, cellular metabolic processes, and substance binding. Under acetic acid stress, strains might attenuate the toxicity of acetic acid by intensifying fatty acid metabolism, weakening the tricarboxylic acid cycle, glycerophospholipid and energy metabolism during the lag phase, while strains might promote the assimilation of acetic acid and inter-conversion of substances during the exponential phase by enhancing the tricarboxylic acid cycle, glycolysis, pyruvate, and energy metabolism to produce and tolerate acid. Besides, cell cycle regulation and protein translation might be potential acid tolerance pathways under high acid stress. The result contributes to the exploration of new potential acid tolerance mechanisms in Acetobacter pasteurianus from four-dimensional label-free relative quantitative proteomics analysis.

In this study, spore-forming bacteria isolated from saccharified rice were selected for producing acetic acid. From the screening of 15 strains, P8 strain was chosen as a candidate. The strain was identified as Paenibacillus azoreducens by 16S rRNA analysis (99.85% similarity with P. azoreducens CM1T). Acetic acid is the main component of vinegar but also an industrial commodity produced by chemical synthesis. Sustainable routes for obtaining acetic acid are of great interest for decreasing the environmental impact generated by chemical syntheses. Biological acetic acid production is effective for vinegar production by acetic acid bacteria, but it cannot economically compete with the chemical synthesis for producing it as a pure commodity. Considering the need to improve the yield of pure acetic acid produced by microbial conversions, in this study, P8 strain was chosen for designing processes in different fermentation conditions. Tests were conducted in single and semi-continuous systems, using rice wine as substrate. Acetic acid produced by P8 strain was compared with that of Acetobacter pasteurianus (UMCC 2951), a strain known for producing acetic acid from rice wine. Even though the fermentation performances of P. azoreducens P8 were slightly lower than those of acetic acid bacteria usually used for vinegar production, results highlight its suitability for producing acetic acid. The final acetic acid produced by P. azoreducens P8 was 73g/L, in a single stage fermentation, without losses. In nine cycles of semi-continuous regime the average of acetification rate was 0.814 (g/L/days). Two main attributes of P. azoreducens P8 are of relevance for producing acetic acid, namely the ability to grow at temperature higher (+ 37°C), than mesophilic acetic acid bacteria, and the absence of cytoplasmic assimilation of acetic acid. These features allow to design multiple strains cultures, in which P. azoreducens can acts as a helper strain. Based on our results, the new isolate P. azoreducens P8 can be propagated in fermenting broths for boosting acetic acid production, under the selected conditions, and used in combination with acetic acid bacteria to produce biological acetic acid, as a non-food grade commodity. [ABSTRACT FROM AUTHOR]

The production of vinegar on an industrial scale from different raw materials is subject to constraints, notably the low tolerance of acetic acid bacteria (AAB) to high temperatures and high ethanol concentrations. In this study, we used 25 samples of different fruits from seven Moroccan biotopes with arid and semi-arid environmental conditions as a basic substrate to isolate thermo- and ethanol-tolerant AAB strains. The isolation and morphological, biochemical and metabolic characterization of these bacteria allowed us to isolate a total number of 400 strains with characters similar to AAB, of which six strains (FAGD1, FAGD10, FAGD18 and GCM2, GCM4, GCM15) were found to be mobile and immobile Gram-negative bacteria with ellipsoidal rod-shaped colonies that clustered in pairs and in isolated chains. These strains are capable of producing acetic acid from ethanol, growing on peptone and oxidizing acetate to CO2 and H2O. Strains FAGD1, FAGD10 and FAGD18 show negative growth on YPG medium containing D-glucose > 30%, while strains GCM2, GCM4 and GCM15 show positive growth. These six strains stand out on CARR indicator medium as isolates of the genus Acetobacter ssp. Analysis of 16S rDNA gene sequencing allowed us to differentiate these strains as Acetobacter fabarum and Acetobacter pasteurianus. The study of the tolerance of these six isolates towards pH showed that most of the six strains are unable to grow at pH 3 and pH 9, with an ideal pH of 5. The behavior of the six strains at different concentrations of ethanol shows an optimal production of acetic acid after incubation at concentrations between 6% and 8% (v/v) of ethanol. All six strains tolerated an ethanol concentration of 16% (v/v). The resistance of the strains to acetic acid differs between the species of AAB. The optimum acetic acid production is obtained at a concentration of 1% (v/v) for the strains of FAGD1, FAGD10 and FAGD18, and 3% (v/v) for GCM2, GCM4 and GCM15. These strains are able to tolerate an acetic acid concentration of up to 6% (v/v). The production kinetics of the six strains show the highest levels of growth and acetic acid production at 30 °C. This rate of growth and acetic acid production is high at 35 °C, 37 °C and 40 °C. Above 40 °C, the production of acid is reduced. All six strains continue to produce acetic acid, even at high temperatures up to 48 °C. These strains can be used in the vinegar production industry to minimize the load on cooling systems, especially in countries with high summer temperatures. [ABSTRACT FROM AUTHOR]

Introduction: Weak aerobic stability is a notable challenge for whole-plant corn silage, particularly in hot and humid regions. Acetobacter is commonly regarded as an indicator of aerobic deterioration in silage, yet its precise role in fermentation and during aerobic exposure, as well as the factors that promote its growth, remain insufficiently understood., Methods: In this study, whole-plant corn silage was prepared using a bagged method with controlled dry matter (DM) content at 20%, 25%, and 30%, and initial concentrations of A. pasteurianus at 40%, 50%, and 60%. The silage was stored for 60 days under varying temperatures (20°C, 30°C, and 40°C). Following the anaerobic storage phase, the silage was exposed to air at room temperature (20-25°C) for 7 days, both with and without A. pasteurianus inoculation., Results: The results demonstrated that A. pasteurianus did not impact the nutritional value of the silage during anaerobic fermentation, maintaining a low pH (< 3.80). However, during aerobic exposure, the presence of A. pasteurianus significantly reduced the aerobic stability of the silage. The microbial community shifted from primarily Klebsiella species initially to Lactobacillus and Acetobacter species post-ensiling. During the aerobic exposure phase, A. pasteurianus and A. fabarum became the dominant species. Response Surface Methodology (RSM) analysis identified optimal conditions for the proliferation of A. pasteurianus during the aerobic phase, which occurred at 28°C, 25% DM, and 52% initial concentration at 3 ml/kg., Discussion: These findings confirm that A. pasteurianus plays a critical role in reducing the aerobic stability of whole-plant corn silage. Additionally, the study identifies the optimal conditions that favor the proliferation of A. pasteurianus , offering valuable insights for the development of strategies to prevent and control this bacterium, thereby improving the aerobic stability of silage in hot and humid regions., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Bai, Li, Chen, Yuan, Chen, Huang, Zhou and Guan.)

In this study, spore-forming bacteria isolated from saccharified rice were selected for producing acetic acid. From the screening of 15 strains, P8 strain was chosen as a candidate. The strain was identified as Paenibacillus azoreducens by 16S rRNA analysis (99.85% similarity with P. azoreducens CM1T). Acetic acid is the main component of vinegar but also an industrial commodity produced by chemical synthesis. Sustainable routes for obtaining acetic acid are of great interest for decreasing the environmental impact generated by chemical syntheses. Biological acetic acid production is effective for vinegar production by acetic acid bacteria, but it cannot economically compete with the chemical synthesis for producing it as a pure commodity. Considering the need to improve the yield of pure acetic acid produced by microbial conversions, in this study, P8 strain was chosen for designing processes in different fermentation conditions. Tests were conducted in single and semi-continuous systems, using rice wine as substrate. Acetic acid produced by P8 strain was compared with that of Acetobacter pasteurianus (UMCC 2951), a strain known for producing acetic acid from rice wine. Even though the fermentation performances of P. azoreducens P8 were slightly lower than those of acetic acid bacteria usually used for vinegar production, results highlight its suitability for producing acetic acid. The final acetic acid produced by P. azoreducens P8 was 73 g/L, in a single stage fermentation, without losses. In nine cycles of semi-continuous regime the average of acetification rate was 0.814 (g/L/days). Two main attributes of P. azoreducens P8 are of relevance for producing acetic acid, namely the ability to grow at temperature higher (+ 37°C), than mesophilic acetic acid bacteria, and the absence of cytoplasmic assimilation of acetic acid. These features allow to design multiple strains cultures, in which P. azoreducens can acts as a helper strain. Based on our results, the new isolate P. azoreducens P8 can be propagated in fermenting broths for boosting acetic acid production, under the selected conditions, and used in combination with acetic acid bacteria to produce biological acetic acid, as a non-food grade commodity.

The microbial community plays an important role on the solid-state fermentation (SSF) of Chinese cereal vinegar, where acetic acid bacteria (AAB) and lactic acid bacteria (LAB) are the dominant bacteria. In this study, the top-down (in situ) and bottom-up (in vitro) approaches were employed to reveal the interaction of AAB and LAB in SSF of Shanxi aged vinegar (SAV). The results of high-throughput sequencing indicates that Acetobacter pasteurianus and Lactobacillus helveticus are the predominant species of AAB and LAB, respectively, and they showed negative interrelationship during the fermentation. A. pasteurianus CGMCC 3089 and L. helveticus CGMCC 12062, both of which were isolated from fermentation of SAV, showed no nutritional competition when they were co-cultured in vitro. However, the growth and metabolism of L. helveticus CGMCC 12062 were inhibited during SSF due to the presence of A. pasteurianus CGMCC 3089, indicating an amensalism phenomenon between these two species. The transcriptomic results shows that there are 831 differentially expressed genes (|log2 (Fold Change)| > 1 and, p ≤ 0.05) in L. helveticus CGMCC 12062 under co-culture condition comparing to its mono-culture, which are mainly classified into Gene Ontology classification of molecular function, biological process, and cell composition. Of those 831 differentially expressed genes, 202 genes are up-regulated and 629 genes are down-regulated. The down-regulated genes were enriched in KEGG pathways of sugar, amino acid, purine, and pyrimidine metabolism. The transcriptomic results for A. pasteurianus CGMCC 3089 under co-culture condition reveals 529 differentially expressed genes with 393 up-regulated and 136 down-regulated, and the genes within KEGG pathways of sugar, amino acid, purine, and pyrimidine metabolism are up-regulated. Results indicate an amensalism relationship in co-culture of A. pasteurianus and L. helveticus. Therefore, this work gives a whole insight on the interaction between the predominant species in SSF of cereal vinegar from nutrient utilization, endogenous factors inhibition and the regulation of gene transcription.

Polyphenols are important functional substances produced in the acetic acid fermentation (AAF) of Shanxi aged vinegar (SAV). Previous studies have shown that the metabolic activity of microorganisms is closely related to polyphenol production and accumulation. In this study, microorganisms in the AAF of SAV were analyzed to explore how to increase the polyphenol yield by changing the microorganisms and reveal the potential mechanism of the microbial influence on the polyphenol yield. Macrotranscriptome analysis showed that acetic and lactic acid bacteria dominated the AAF fermentation process and initially increased and decreased. Spearman correlation analysis and verification experiments showed that the co-addition of Acetobacter pasteurianus and Lactobacillus helveticus promoted the accumulation of polyphenols, and the total polyphenol content increased by 72% after strengthening.

This study investigated the influence of one- and two-step fermentation on bioactive compound production in fermented green tea, i.e., one-step fermented green tea (OFG) and two-step fermented green tea (TFG). One-step fermentation entailed acetic acid fermentation, while two-step fermentation consisted of lactic acid fermentation followed by acetic acid fermentation. Acetobacter pasteurianus PCH 325, isolated from an over-ripened peach, was selected for acetic acid fermentation based on its growth and organic acid production characteristics. Acetic acid fermentation conditions were optimized for one- and two-step fermentation: 3% fermentation alcohol for both processes; 8% and 4% sucrose, respectively; and fermentation at 25 ◦C for both processes. For lactic acid fermentation of TFG, the inoculum and optimized conditions reported previously were used. Under the optimized conditions, the acetic acid content in OFG and TFG increased 21.20- and 29.51-fold, respectively. Furthermore, through two-step fermentation, γ-aminobutyric acid and lactic acid were produced up to 31.49 ± 1.17 mg/L and 243.44 ± 58.15 mg/L, respectively, which together with acetic acid could contribute to the higher DPPH scavenging activity of TFG. This study suggests that two-step fermentation may be a valuable strategy in industry for raising the amount of acetic acid and/or providing additional bioactive compounds. [ABSTRACT FROM AUTHOR]

The objective of this study is to explore the effect and mechanism of ultrasound on chitin extraction from shrimp shells powder (SSP) by the co-fermentation of Bacillus subtilis and Acetobacter pasteurianus. After pre-treating the SSP with high-intensity ultrasound (HIU) at 800 W, the protease activity in the fermentation solution reached 96.9 U/mL on day 3, which was significantly higher than for SSP that had not been pre-treated with ultrasound (81.8 U/mL). The fermentation time of the chitin extraction process was 5.0 d without ultrasound pre-treatment, while it was shortened to 4.5 d when using ultrasound at 800 W to treat SSP. However, there were no obvious differences when we applied ultrasound at low power (200 W, 400 W). Furthermore, chitin purified from shrimp shells pre-treated with HIU at 800 W exhibited lower molecular weight (11.2 kDa), higher chitin purity (89.8%), and a higher degree of deacetylation (21.1%) compared to SSP with no ultrasound pre-treatment (13.5 kDa, 86.6%, 18.5%). Results indicate that HIU peels off the protein/CaCO3 matrix that covers the SSP surface. About 9.1% of protein and 4.7% of Ca2+ were released from SSP pre-treated with HIU at 800 W. These figures were both higher than with no ultrasound pre-treatment (4.5%, 3.2%). Additionally, the amount of soluble protein extracted from SSP through HIU at 800 W was 50% higher than for the control sample. SDS-PAGE analysis indicated that the soluble protein was degraded to the micromolecule. It also revealed that HIU (600, 800 W) induced the secondary and tertiary structure destruction of protein extracted from SSP. In conclusion, HIU-induced degradation and structural damage of protein enhances the protein/CaCO3 matrix to be peeled off from SSP. Also, in the co-fermentation process, an increase of protease activity further accelerates deproteinization.

To investigate the quality variations of prune vinegar during fermentation, multivariate statistical analysis (MSA) was used in conjunction with high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS). The results showed that the pH decreased slightly, there was a rapid decline in sugar content, and a gradual increase in total acid content, which achieved stabilization at 3.68 g/100 mL. The content of organic acid increased, while the total phenol content decreased. The total flavonoid content did not change significantly. Furthermore, the antioxidant capacity initially decreased and then increased. Prune vinegar contains important phenolics such as rutin, resveratrol, salicylic acid, trans -ferulic acid, and (+)-catechin. In addition, the flavor of prune fruit vinegar underwent changes during the fermentation process. During the fermentation of fruit vinegar, large quantities of 2,3-butanediol, glacial acetic acid, nonanoic acid, ethyl acetate, ethyl heptanoate, ethyl caproate, and linalool were produced. These compounds give prune vinegar its distinctive fruity and floral aroma. This study provided a comprehensive framework for the efficient processing of prunes and a strategy for enhancing the quality characteristics of prune vinegar or other fermented products. • Determination of phenolic compounds in prune vinegar. • Prune vinegar has high antioxidant activity. • Fermented prune vinegar produces more unique aromatic substances. • The major volatile compounds of prune vinegar were identified. [ABSTRACT FROM AUTHOR]

Traditional grain vinegar is fermented using multiple acetic acid bacteria (AAB) at various temperatures. A single AAB showed high acid-producing ability at 30 °C with a 5% alcohol concentration and an initial pH adjusted to 4.0. Multiple AAB were similar to a single AAB; however, the optimal initial pH was 3.0. Acid production ability according to the type of AAB was higher in multiple AAB than in single AAB. That is, using multiple AAB helped increase the titratable acidity of traditional grain vinegar. In addition, increasing the titratable acidity and content of volatile flavor compounds was advantageous when two, rather than four, AAB types were mixed and used. The titratable acidity was high at medium temperatures (30 °C); however, volatile flavor compounds increased at low temperatures (20 °C) under multiple AAB. A 16S rDNA-based microbiome taxonomic profiling analysis identified differences in beta diversity due to multiple AAB and fermentation temperatures. In particular, beta diversity analysis revealed a specific pattern when a mixture of Acetobacter ascedens GV–8 and Acetobacter pasteurianus GV–22 was fermented at a low temperature (20 °C). Therefore, we propose the application of multiple AAB with acidic and flavor-producing properties in traditional grain vinegar.

2,3-Butanediol (2,3-BD) has been extensively used in chemical syntheses. This study aimed to explore acetic acid as a signaling molecule that activates a quorum sensing (QS) system to promote the production of 2,3-BD. The yield of 2,3-BD is proportional to the cell density. Saccharomyces cerevisiae W141 does not produce 2,3-BD when the cell density is lower than the threshold concentration (OD600 nm = 10 or cell density 4.4 × 108 CFU/mL). When 1.5 g/L acetic acid is added, the yield of 2,3-BD is 3.01 ± 0.04 g/L. Subsequently, S. cerevisiae W141 was cocultured with Acetobacter pasteurianus Huniang 1.01 under the optimal conditions, the acetic acid production was increased by 76.7% and 30.6% compared with the original strain and the strain cultivated with 1.5 g/L acetic acid, and the yield of 2,3-BD was increased by 81.9% and 3.3%, respectively. This difference is due to the activity of acetyl lactic acid synthase (ILV2) and 2,3-BD dehydrogenase (BDH1), as the relative expression of the ilv2 and bdh1 genes is increased. The results showed that the biosynthesis of 2,3-BD was regulated by acetic acid as a signaling molecule. S. cerevisiae is a promising host for producing 2,3-BD for industrial applications. [ABSTRACT FROM AUTHOR]

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Energy metabolism is important for cell growth and tolerance against environment stress. In acetic acid fermentation by Acetobacter pasteurianus, the correlation coefficients of acid production rate with energy charge and ATP content were 0.9981 and 0.9826, respectively. The main energy metabolism pathway, including glycolysis pathway, TCA cycle, ethanol oxidation, pentose phosphate pathway, and ATP production, was constructed by transcriptome analysis. The effects of fermentation conditions, including dissolved oxygen, initial acetic acid concentration, and total concentration, on acetic acid fermentation and energy metabolism of A. pasteurianus were analyzed by using the RT-PCR method. The results showed the high energy charge inhibited glucose catabolism, and associated with the high ethanol oxidation rate. Consequently, a virtuous circle of increased ethanol oxidation, increased energy generation, and acetic acid tolerance was important for improving acetic acid fermentation.

Exposure to ionizing radiation (IR) tends to cause serious health concerns. Thus, radioprotective agents are vital for the population exposed to radiation. As microorganisms have the advantages of fast reproduction and no geographical restrictions, direct microbe-based and environmental induction compounds are thriving radioprotectants resources. Oxidative system and oxidase in Acetobacter pasteurianus are unique and intriguing, the radioprotective effect of the cell-free extract from A. pasteurianus (APE) and 60Coγ-treated extract (IRE) were comparatively investigated in the present study. The survival rate of A. pasteurianus with IRE addition was 149.1% in H2O2 damage test, while that with APE was only 10.4%. The viability of 60Coγ-treated AML-12 cells was increased by 18.8% with IRE addition, yet APE showed no significant radioprotective effect. Moreover, in 60Coγ-treated mice, IRE could significantly protect the white blood cell, improve the liver index, and attenuate the injuries of immune organs in mice. Administration of IRE significantly raised the activities of superoxide dismutase (SOD) and reduced the products of lipid peroxidation. These results clarified that gavage with APE and IRE presented notable antioxidant and radioprotective efficacy. A. pasteurianus showed appealing potential to be novel radioprotective bioagents and 60Coγ treatment on microbe could be a new method for the development of better radioprotectant. Key points: • 60Coγ induction could improve the radioprotective effect of APE. • IRE protected white blood cell in mice under IR. • IRE products have broad application prospects in radioprotection based on microbes. [ABSTRACT FROM AUTHOR]

Abstract Excessive alcohol consumption induces acute intoxication and various hepatic diseases. In this study, we investigated the effect of the CureZyme-ACE (CA), Acetobacter Pasteurianus (AP)-derived product, in acute intoxication rats. The ethanol and acetaldehyde levels of serum were lower in rats treated with CA than those who only treated ethanol. The activities of alcohol dehydrogenase and acetaldehyde dehydrogenase also recovered faster in the CA group than only-ethanol group. The transaminase levels (AST, ALT) in the CA group were significantly lower than only-ethanol group. In addition, Hepatic histological analyses and stomach wall were demonstrated that the CA-treated group recovered faster than only-ethanol group. With regard to most characteristics, we found that CA had dose-dependent effects. At high concentrations of CA, there were no differences in the tested parameters compared to those of normal rats. These findings indicate that CA reduces the serum alcohol concentration and some of the hepatic damage caused by alcohol intoxication.

Copyright of China Brewing is the property of China Brewing Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

This study aims to develop a method for the preparation of natural table olives using locally selected microorganisms and without resorting to the usual techniques which employ lye treatment and acids. The effects of parameters, such as lye treatment, inoculation with yeasts, substitution of organic acids with vinegar and/or acetic acid bacteria, and finally alternating aeration have been assessed. Four different combinations were applied to the “Picholine marocaine” olive variety using indigenous strains, namely Lactobacillus plantarum S1, Saccharomyces cerevisiae LD01 and Acetobacter pasteurianus KU710511 (CV01) isolated respectively from olive brine, Bouslikhen dates and Cactus. Two control tests, referring to traditional and industrial processes, were used as references. Microbial and physicochemical tests showed that the L3V combination (inoculated with A. pasteurianus KU710511 and L. plantarum S1 under the optimal growth conditions of the Acetic Acid Bacteria (AAB) strain with 6% NaCl) was found to be favorable for the growth of the Lactic Acid Bacteria (LAB) strain which plays the key role in olive fermentation. This result was confirmed by sensory evaluation, placing L3V at the top of the evaluated samples, surpassing the industrial one where a chemical debittering treatment with lye was used. In addition, alternating aeration served to increase the microbial biomass of both AAB and LAB strains along with Saccharomyces cerevisiae LD01 strain, but also to use lower concentration of NaCl and to reduce the deterioration of olives compared to the anaerobic fermentation process. Finally, a mixed starter containing the three strains was prepared in a 10-L Lab-fermenter from the L3V sample in order to improve it in subsequent studies. The prepared starter mixture could be suitable for use as a parental strain to prepare table olives for artisan and industrial application in Morocco.

Acetic acid bacteria (AAB) are a group of Gram-negative and strictly aerobic microorganisms widely used in vinegar industry, especially the species belonging to the genera Acetobacter and Komagataeibacter. The environments inhabited by AAB during the vinegar fermentation, in particular those natural traditional bioprocesses, are complex and dynamically changed, usually accompanied by diverse microorganisms, bacteriophages, and the increasing acetic acid concentration. For this reason, how AAB survive to such harsh niches has always been an interesting research field. Previous omic analyses (e.g., genomics, proteomics, and transcriptomics) have provided abundant clues for the metabolic pathways and bioprocesses indispensable for the acid stress adaptation of AAB. Nevertheless, it is far from fully understanding what factors regulate these modular mechanisms overtly and covertly upon shifting environments. Bacterial toxin-antitoxin systems (TAS), usually consisting of a pair of genes encoding a stable toxin and an unstable antitoxin that is capable of counteracting the toxin, have been uncovered to have a variety of biological functions. Recent studies focusing on the role of TAS in Acetobacter pasteurianus suggest that TAS contribute substantially to the acid stress resistance. In this mini review, we discuss the biological functions of type II TAS in the context of AAB with regard to the acid stress resistance, persister formation and resuscitation, genome stability, and phage immunity. Key points: • Type II TAS act as regulators in the acid stress resistance of AAB. • Type II TAS are implicated in the formation of acid-tolerant persister cells in AAB. • Type II TAS are potential factors responsible for phage immunity and genome stability. [ABSTRACT FROM AUTHOR]

In the present study, we aimed to determine the cause of surface film formation in three rice vinegars fermented using the traditional static fermentation method. The pH and total acidity of vinegar were 3.0–3.3 and 3.0–8.7%, respectively, and acetic acid was the predominant organic acid present. Colonies showing a clear halo on GYC medium were isolated from the surface film of all vinegars. Via 16S rDNA sequencing, all of the isolates were identified as Acetobacter pasteurianus. Furthermore, field-emission scanning electron microscopy analysis showed that the bacterial cells had a rough surface, were rod-shaped, and were ∼1 × 2 µm in size. Interestingly, cells of the isolate from one of the vinegars were surrounded with an extremely fine threadlike structure. Thus, our results suggest that formation of the surface film in rice vinegar was attributable not to external contamination, to the production of bacterial cellulose by A. pasteurianus to withstand the high concentrations of acetic acid generated during fermentation. However, because of the formation of a surface film in vinegar is undesirable from an industrial perspective, further studies should focus on devising a modified fermentation process to prevent surface film formation and consequent quality degradation.

The production of vinegar on an industrial scale from different raw materials is subject to constraints, notably the low tolerance of acetic acid bacteria (AAB) to high temperatures and high ethanol concentrations. In this study, we used 25 samples of different fruits from seven Moroccan biotopes with arid and semi-arid environmental conditions as a basic substrate to isolate thermo- and ethanol-tolerant AAB strains. The isolation and morphological, biochemical and metabolic characterization of these bacteria allowed us to isolate a total number of 400 strains with characters similar to AAB, of which six strains (FAGD1, FAGD10, FAGD18 and GCM2, GCM4, GCM15) were found to be mobile and immobile Gram-negative bacteria with ellipsoidal rod-shaped colonies that clustered in pairs and in isolated chains. These strains are capable of producing acetic acid from ethanol, growing on peptone and oxidizing acetate to CO2 and H2O. Strains FAGD1, FAGD10 and FAGD18 show negative growth on YPG medium containing D-glucose > 30%, while strains GCM2, GCM4 and GCM15 show positive growth. These six strains stand out on CARR indicator medium as isolates of the genus Acetobacter ssp. Analysis of 16S rDNA gene sequencing allowed us to differentiate these strains as Acetobacter fabarum and Acetobacter pasteurianus. The study of the tolerance of these six isolates towards pH showed that most of the six strains are unable to grow at pH 3 and pH 9, with an ideal pH of 5. The behavior of the six strains at different concentrations of ethanol shows an optimal production of acetic acid after incubation at concentrations between 6% and 8% (v/v) of ethanol. All six strains tolerated an ethanol concentration of 16% (v/v). The resistance of the strains to acetic acid differs between the species of AAB. The optimum acetic acid production is obtained at a concentration of 1% (v/v) for the strains of FAGD1, FAGD10 and FAGD18, and 3% (v/v) for GCM2, GCM4 and GCM15. These strains are able to tolerate an acetic acid concentration of up to 6% (v/v). The production kinetics of the six strains show the highest levels of growth and acetic acid production at 30 °C. This rate of growth and acetic acid production is high at 35 °C, 37 °C and 40 °C. Above 40 °C, the production of acid is reduced. All six strains continue to produce acetic acid, even at high temperatures up to 48 °C. These strains can be used in the vinegar production industry to minimize the load on cooling systems, especially in countries with high summer temperatures.

Thermotolerant microorganisms are useful for high-temperature fermentation. Several thermally adapted strains were previously obtained from Acetobacter pasteurianus in a nutrient-rich culture medium, while these adapted strains could not grow well at high temperature in the nutrient-poor practical culture medium, "rice moromi." In this study, A. pasteurianus K-1034 originally capable of performing acetic acid fermentation in rice moromi was thermally adapted by experimental evolution using a "pseudo" rice moromi culture. The adapted strains thus obtained were confirmed to grow well in such the nutrient-poor media in flask or jar-fermentor culture up to 40 or 39 °C; the mutation sites of the strains were also determined. The high-temperature fermentation ability was also shown to be comparable with a low-nutrient adapted strain previously obtained. Using the practical fermentation system, "Acetofermenter," acetic acid production was compared in the moromi culture; the results showed that the adapted strains efficiently perform practical vinegar production under high-temperature conditions. [ABSTRACT FROM AUTHOR]

This study aims to develop a method for the preparation of natural table olives using locally selected microorganisms and without resorting to the usual techniques which employ lye treatment and acids. The effects of parameters, such as lye treatment, inoculation with yeasts, substitution of organic acids with vinegar and/or acetic acid bacteria, and finally alternating aeration have been assessed. Four different combinations were applied to the "Picholine marocaine" olive variety using indigenous strains, namely Lactobacillus plantarum S1, Saccharomyces cerevisiae LD01 and Acetobacter pasteurianus KU710511 (CV01) isolated respectively from olive brine, Bouslikhen dates and Cactus. Two control tests, referring to traditional and industrial processes, were used as references. Microbial and physicochemical tests showed that the L3V combination (inoculated with A. pasteurianus KU710511 and L. plantarum S1 under the optimal growth conditions of the Acetic Acid Bacteria (AAB) strain with 6% NaCl) was found to be favorable for the growth of the Lactic Acid Bacteria (LAB) strain which plays the key role in olive fermentation. This result was confirmed by sensory evaluation, placing L3V at the top of the evaluated samples, surpassing the industrial one where a chemical debittering treatment with lye was used. In addition, alternating aeration served to increase the microbial biomass of both AAB and LAB strains along with Saccharomyces cerevisiae LD01 strain, but also to use lower concentration of NaCl and to reduce the deterioration of olives compared to the anaerobic fermentation process. Finally, a mixed starter containing the three strains was prepared in a 10-L Lab-fermenter from the L3V sample in order to improve it in subsequent studies. The prepared starter mixture could be suitable for use as a parental strain to prepare table olives for artisan and industrial application in Morocco. [ABSTRACT FROM AUTHOR]

Acetobacter pasteurianus accumulates reactive oxygen species (ROS). ROS are produced by electron and oxygen coupling in the electron transport chain in the intracellular environment during the stationary and in the acetic acid over-oxidation phases in the presence of ethanol, thereby exposing cell to oxidative stress. In this study, to reveal the resistance mechanism to oxidative stress in A. pasteurianus , we focused on DNA-binding protein from starvation cells (Dps) and analyzed the function of Dps against oxidative stress. When Dps under the copresence of plasmid DNA was exposed to H 2 O 2 and divalent iron, plasmid DNA fragmentation was suppressed under the presence of Dps; however, DNA binding was not observed, revealing a defensive activity for oxidative damage. In addition, this finding revealed that Dps incorporates a divalent iron intracellularly, forming a ferroxidase center. Moreover, levels of hydroxyl radicals produced by Fenton reaction under the presence of H 2 O 2 and divalent iron were decreased by the addition of Dps, resulting in the suppression of the Fenton reaction. Through fluorescence microscopy using a divalent-iron-specific fluorescent probe, we found that, in dps gene disruptants, the accumulation of the divalent iron increased, and the dps gene disruptants showed higher sensitivity to H 2 O 2 than the wild-type. These result strongly suggested that Dps traps intracellular free-divalent iron and plays an important role in the oxidative stress resistance of A. pasteurianus NBRC 3283 after the acetic acid fermentation phase. [ABSTRACT FROM AUTHOR]

Vinegar is a liquid product produced from alcoholic and acetous fermentation. Vinegar contains acetic acid and bioactive compounds, which are brewed by liquid-state and solid-state fermentation techniques. This study reviews chemical fermentation process properties, bioactive compounds, and health benefits of fruit vinegars in Thailand. Chemical properties are the alcohol percentage and acetic acid percentage of vinegar. Bioactive compounds include antioxidant activity and total phenolic content, which have the role of antioxidative activity, blood pressure and glucose control, and anti-tumor. However, further studies are needed to find the new fruit raw materials to produce fruit vinegar which have more bioactive compound and more good taste. [ABSTRACT FROM AUTHOR]

Elucidation of the acetic acid resistance (AAR) mechanisms is of great significance to the development of industrial microbial species, specifically to the acetic acid bacteria (AAB) in vinegar industry. Currently, the role of population heterogeneity in the AAR of AAB is still unclear. In this study, we investigated the persister formation in AAB and the physiological role of HicAB in Acetobacter pasteurianus Ab3. We found that AAB were able to produce a high level of persister cells (10−2 to 100 in frequency) in the exponential-phase cultures. Initial addition of acetic acid and ethanol reduced the ratio of persister cells in A. pasteurianus by promoting the intracellular ATP level. Further, we demonstrated that HicAB was an important regulator of AAR in A. pasteurianus Ab3. Strains lacking hicAB showed a decreased survival under acetic acid exposure. Deletion of hicAB significantly diminished the acetic acid production, acetification rate, and persister formation in A. pasteurianus Ab3, underscoring the correlation between hicAB, persister formation, and acid stress resistance. By transcriptomic analysis (RNA-seq), we revealed that HicAB contributed to the survival of A. pasteurianus Ab3 under high acid stress by upregulating the expression of genes involved in the acetic acid over-oxidation and transport, 2-methylcitrate cycle, and oxidative phosphorylation. Collectively, the results of this study refresh our current understanding of the AAR mechanisms in A. pasteurianus, which may facilitate the development of novel ways for improving its industrial performance and direct the scaled-up vinegar production. Key points: • AAB strains form persister cells with different frequencies. • A. pasteurianus are able to form acid-tolerant persister cells. • HicAB contributes to the AAR and persister formation in A. pasteurianus Ab3. [ABSTRACT FROM AUTHOR]

Acetic acid accumulation is a universal limiting factor to the vinegar manufacture because of the toxic effect of acetic acid on the acid producing strain, such as Acetobacter pasteurianus. In this study, we aimed to investigate the genome-wide transcriptional response of A. pasteurianus Ab3 to high acid stress during vinegar production. By comparing the transcriptional landscape of cells harvested from a long-term cultivation with high acidity (70 ± 3 g/L) to that of low acidity (10 ± 2 g/L), we demonstrated that 1005 genes were differentially expressed. By functional enrichment analysis, we found that the expression of genes related to the two-component systems (TCS) and toxin-antitoxin systems (TAS) was significantly regulated under high acid stress. Cells increased the genome stability to withstand the intracellular toxicity caused by the acetic acid accumulation by repressing the expression of transposases and integrases. Moreover, high acid stress induced the expression of genes involved in the pathways of peptidoglycan, ceramide, and phosphatidylcholine biosynthesis as well as the Tol-Pal and TonB-ExbB systems. In addition, we observed that cells increased and diversified the ATP production to resist high acid stress. Transcriptional upregulation in the pathways of pyrroloquinoline quinone (PQQ) synthesis and thiamine metabolism suggested that cells may increase the production of prosthetic groups to ensure the enzyme activity upon high acid stress. Collectively, the results of this study increase our current understanding of the acetic acid resistance (AAR) mechanisms in A. pasteurianus and provide opportunities for strain improvement and scaled-up vinegar production. Key Points • TCS and TAS are responsive to the acid stress and constitute the regulating networks. • Adaptive expression changes of cell envelope elements help cell resist acid stress. • Cells promote genome stability and diversify ATP production to withstand acid stress. [ABSTRACT FROM AUTHOR]

Bacterial cellulose has unique structural, functional, physical and chemical properties. The mass production of bacterial cellulose for industrial application has recently become more popular and attractive. The aim of this study was to develop an efficient scale-up process of bacterial cellulose production. Bacterial cellulose productivity of a new isolate Acetobacter pasteurianus RSV-4 (MTCC 25117) was investigated in whey medium without addition of any additives. Produced bacterial cellulose was characterized for its structure, purity, thermostability and strength by Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, thermogravimetric analysis and Differential scanning calorimetry. β-galactosidase (1.5 IU/ml) was used to split whey lactose into its corresponding monomers glucose and galactose. Glucose was specifically consumed by Acetobacter pasteurianus for the production of BC. The highest concentration of 5.6 g cellulose/L of whey was obtained at 30 °C after 8 days of bacterial growth. Process was optimized for the production of bacterial cellulose on 120 L of whey medium under static conditions. Whey, a waste byproduct of milk processing industry was used in the process and therefore the developed process might be economical. [ABSTRACT FROM AUTHOR]

Acetobacter pasteurianus 386B is a candidate functional starter culture for the cocoa bean fermentation process. To allow in silico simulations of its related metabolism in response to different environmental conditions, a genome-scale metabolic model for A. pasteurianus 386B was reconstructed. This is the first genome-scale metabolic model reconstruction for a member of the genus Acetobacter. The metabolic network reconstruction process was based on extensive genome re-annotation and comparative genomics analyses. The information content related to the functional annotation of metabolic enzymes and transporters was placed in a metabolic context by exploring and curating a Pathway/Genome Database of A. pasteurianus 386B using the Pathway Tools software. Metabolic reactions and curated gene-protein-reaction associations were bundled into a genome-scale metabolic model of A. pasteurianus 386B, named iAp386B454, containing 454 genes, 322 reactions, and 296 metabolites embedded in two cellular compartments. The reconstructed model was validated by performing growth experiments in a defined medium, which revealed that lactic acid as the sole carbon source could sustain growth of this strain. Further, the reconstruction of the A. pasteurianus 386B genome-scale metabolic model revealed knowledge gaps concerning the metabolism of this strain, especially related to the biosynthesis of its cell envelope and the presence or absence of metabolite transporters.

Elucidation of the acetic acid resistance (AAR) mechanisms of Acetobacter pasteurianus is significant for vinegar production. In this study, cell membrane lipid profile of A. pasteurianus Ab3 was investigated by gas chromatography-mass spectrometer (GC–MS) and high performance liquid chromatography–electrospray ionization (HPLC-ESI) combined with high resolution accurate mass/mass spectrometry (HRAM/MS). We observed that cell remodeled the membrane physical state by decreasing the ratio of saturated fatty acids (SFAs)/unsaturated fatty acids (UFAs), and increasing the chain length of fatty acids (FAs) and the content of cyclopropane FAs in response to extreme acid stress. Noticeably, the content of octadecadienoic acid (C18:2) elevated remarkably. Moreover, a continuous reduction in cell membrane fluidity and a "V-type" variance in permeability were discovered. The content of glycerophospholipid and ceramide increased significantly in cells harvested from culture with acidity of 75 g/L and 95 g/L compared to that with acidity of 30 g/L. Among the identified lipid species, the content of phosphatidylcholine (e.g. PC 19:0/18:2 and 19:1/18:0), ceramide (e.g. Cer d18:0/16:1 and d18:0/16:1 + O), and dimethylphosphatidylethanolamine (e.g. dMePE 19:1/16:1) increased notably with increasing acidity. Collectively, these findings refresh our current understanding of the AAR mechanisms in A. pasteurianus Ab3, and should direct future strain breeding and vinegar fermentation. [ABSTRACT FROM AUTHOR]

• High initial acetic acid-high temperature acetification (HTA) process is explored. • Long adaptation cultivation cycles provide stable acetic acid productions. • CaCl 2 can improve strength of acetic acid bacteria (AAB) cells and activities. • Prototype scale of HTA is tested for solving the major lack in controlled process. • Implementing the HTA respect to the optimal AAB growth, reducing production costs. In this study Acetobacter pasteurianus strain UMCC 2951 was tested as a microbial starter to conduct acetification processes by repeatedly cultivation cycles under high temperature acetification at 40 ± 1 °C. Acid production and acetification rate increased with repeated cultures under high temperature acetification as adaptation period increased, but were still lower than acetification at 30 ± 1 °C. However, the addition of 0.15 % calcium chloride reduced the negative effects of 40 ± 1 °C on both acid production and acetification rate compared to 30 ± 1 °C. A strong decrease in fatty acids and phosphatidylethanolamine and increases in phosphatidylcholine and phosphatidylglycerol in cell membranes were found under high acid and high temperature acetification. In addition, transmission electron microscope images reveal a more compact cell wall when calcium chloride was added to the cultivation medium. The strategy used in this study confirmed that the use of acetic acid bacteria as microbial starters could be effective also at temperature above the optimal values, when acetification processes are managed through repeated semi-continuous cycles. [ABSTRACT FROM AUTHOR]

There is ongoing quest to look for alternate sustainable and renewable biopolymers which can address the existing environmental issues. Bacterial cellulose could be one such option. Several organisms have been reported to produce bacterial cellulose. Among this, acetic acid bacteria (AAB) are reported to be one of the major producers of bacterial cellulose. Recently, we have identified an Acetobacter pasteurianus RSV-4 and reported to produce high tensile strength bacterial cellulose. In order to globally understand its genetic structure, a draft genome sequence of Acetobacter pasteurianus RSV-4 was performed in the present study. The assembled genome had 101 contigs contributing to a total length of 3.8 Mbp. Predicted coding DNA sequences were 3311, of which approximately 70% were assigned the functions. Genome level phylogenetic analysis revealed that RSV-4 belongs to A. pasteurianus. Glycolysis was found to be incomplete in the genome analysis of RSV-4, while the genes/enzymes involved in pentose-phosphate pathway were present. The final draft genome sequence lacked bacterial cellulose synthase (bcs) operon. However, the presence of operon was evident in raw genomic sequences by Sanger sequencing. Therefore, presence of bcs operon in Acetobacter pasteurianus RSV-4 has documented its potential for bacterial cellulose production. • Draft genome of bacterial cellulose producing Acetobacter pasteurianus RSV-4 strain • Total genome size of 3.8 Mbpconsists of 101 contigs and 3311 coding DNA sequences (CDS) • Predicted an operon for bacterial cellulose synthase (bcs) in A. pasteurianus RSV-4 • Predicted bcs operon was further confirmed through Sanger sequencing. [ABSTRACT FROM AUTHOR]

Thermotolerant strains are critical for low-cost high temperature fermentation. In this study, we carried out the thermal adaptation of A. pasteurianus IFO 3283–32 under acetic acid fermentation conditions using an experimental evolution approach from 37ºC to 40ºC. The adapted strain exhibited an increased growth and acetic acid fermentation ability at high temperatures, however, with the trade-off response of the opposite phenotype at low temperatures. Genome analysis followed by PCR sequencing showed that the most adapted strain had 11 mutations, a single 64-kb large deletion, and a single plasmid loss. Comparative phenotypic analysis showed that at least the large deletion (containing many ribosomal RNAs and tRNAs genes) and a mutation of DNA polymerase (one of the 11 mutations) critically contributed to this thermotolerance. The relationship between the phenotypic changes and the gene mutations are discussed, comparing with another thermally adapted A. pasteurianus strains obtained previously. In vitro thermal adaption of mesophilic Acetic Acid Bacteria generates thermally adapted strains exhibiting both thermotolerance and trade-off response [ABSTRACT FROM AUTHOR]

Acetobacter pasteurianus 386B is a candidate functional starter culture for the cocoa bean fermentation process. To allow in silico simulations of its related metabolism in response to different environmental conditions, a genome-scale metabolic model for A. pasteurianus 386B was reconstructed. This is the first genome-scale metabolic model reconstruction for a member of the genus Acetobacter. The metabolic network reconstruction process was based on extensive genome re-annotation and comparative genomics analyses. The information content related to the functional annotation of metabolic enzymes and transporters was placed in a metabolic context by exploring and curating a Pathway/Genome Database of A. pasteurianus 386B using the Pathway Tools software. Metabolic reactions and curated gene-protein-reaction associations were bundled into a genome-scale metabolic model of A. pasteurianus 386B, named iAp386B454, containing 454 genes, 322 reactions, and 296 metabolites embedded in two cellular compartments. The reconstructed model was validated by performing growth experiments in a defined medium, which revealed that lactic acid as the sole carbon source could sustain growth of this strain. Further, the reconstruction of the A. pasteurianus 386B genome-scale metabolic model revealed knowledge gaps concerning the metabolism of this strain, especially related to the biosynthesis of its cell envelope and the presence or absence of metabolite transporters. [ABSTRACT FROM AUTHOR]