Ethanol-tolerant and acetic-tolerant acetobacter pasteurianus and application thereof in production of fruit vinegar

Abstract

The invention discloses an ethanol-tolerant and acetic-tolerant acetobacter pasteurianus strain and application thereof in production of fruit vinegar. The invention provides an acetobacter pasteurianus CL247, the classification name of the acetobacter pasteurianus is Acetobacter pasteurianus, the acetobacter pasteurianus is preserved in China General Microbiological Culture Collection Center (CGMCC) on March 21, 2025, the preservation number is CGMCC No.33924, and the preservation address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Beichen West Road, Chaoyang District, Beijing. The acetobacter pasteurianus CL247 has the characteristics that the acid yield in 72 hours is greater than or equal to 49 g / L, and the acetobacter pasteurianus CL247 can tolerate 7% acetic acid and 18% ethanol. An efficient, tolerant and stable strain solution is provided for fruit vinegar production, the industry is promoted to be upgraded from'low-efficiency discontinuous' to'efficient continuous', and remarkable technical innovation and economic value are achieved.

Description

Technical Field

[0001] The invention relates to an ethanol-resistant and acetate-resistant Acetobacter pasteurianus strain and application thereof in fruit vinegar production. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance some understanding of the overall background of the invention and should not be necessarily regarded as an admission or any form of suggestion that the information constitutes the prior art already known to those skilled in the art.

[0003] The microbial fermentation process of vinegar mainly includes two stages: alcohol fermentation and acetic acid fermentation. Among them, the acetic acid fermentation process mainly depends on Acetobacter pasteurianus ( Acetobacter pasteurianus Acetic acid bacteria, such as acetic acid bacteria, oxidize ethanol to acetic acid (also known as acetic acid). Therefore, during the vinegar production process, acetic acid bacteria are constantly exposed to environmental stresses such as high ethanol and acetic acid concentrations. During fermentation, high concentrations of acetic acid easily penetrate the cell membrane and enter the cytoplasm, adversely affecting bacterial growth and metabolism. Therefore, the environmental stress caused by ethanol and acetic acid is a key limiting factor in the growth and metabolism of acetic acid bacteria. Currently, acetic acid bacteria strains used in industry have significant technical challenges: First, they have low tolerance to ethanol and acetic acid. Most acetic acid bacteria are very sensitive to ethanol. Generally, high ethanol concentrations (>10%) inhibit their growth, resulting in delayed fermentation start-up. When acetic acid concentrations accumulate above 3%, the activity of acetic acid bacteria strains drops sharply, or even ceases to grow. Second, acid production efficiency is low, with acid production often below 45g / L in 72 hours. Third, industrial adaptability is poor. Existing acetic acid bacteria strains struggle to simultaneously withstand extreme conditions such as ethanol and acetic acid, resulting in low production efficiency, high costs, and difficulty meeting the requirements of industrial continuous production.

[0004] While some improved strains have achieved breakthroughs in individual properties, there is a lack of acetic acid bacteria strains that simultaneously exhibit high tolerance to both acetic acid and ethanol (synergistic tolerance to both ethanol and acetic acid). In recent years, improving strain performance through methods such as mutation breeding and gene editing has become a mainstream research direction. For example, Qi Zhengliang et al. selected a high-yielding acetic acid bacterium, Acetobacter pasteurianus CICIM B7003-02, through acid stimulation combined with ultraviolet mutagenesis. This strain can tolerate 6% (60 g / L) acetic acid, with a maximum acid production of 103 g / L. However, this strain's growth is inhibited when the initial ethanol concentration exceeds 48 g / L (Qi Zhengliang. Study on the mechanism of acid stimulation combined with ultraviolet mutagenesis to enhance the acid production capacity and acid tolerance of Acetobacter [D]. Jiangnan University, 2013). Xianle Health's patented strain SP021 can tolerate 14% ethanol and grow at pH 2.0, producing more acid than traditional strains, but its acetic acid tolerance and acid production efficiency are not specified (an ethanol-tolerant, acid-resistant, high-acid-producing Acetobacter pasteurii strain, patent number ZL 202311061245.0). The acetic acid bacteria strain screened by Sun Wenying has an ethanol tolerance of 11%, but its acid resistance is not disclosed, and its final acid production is 48.6 g / L (Sun Wenying, Chen Xiong, Wang Zhi, et al. Screening of high-temperature resistant and high-alcohol acetic acid bacteria and preliminary optimization of their fermentation process [J]. China Brewing, 2011, (01): 44-47). Summary of the Invention

[0005] In response to the shortcomings of the existing technology, the Acetobacter pasteurianus CL247 provided by the present invention is obtained by natural screening from the mash of Shanxi aged vinegar. It achieves balanced breakthroughs in ethanol resistance, acetic acid resistance and acid production efficiency. It is particularly suitable for the continuous production of fruit vinegar with high acetic acid concentration and high ethanol content, and has significant technical advantages and economic value.

[0006] The technical solution adopted in the present invention is as follows: In the first aspect of the present invention, a strain of Acetobacter pasteurianus CL247 is provided, which is classified as Acetobacter pasteurianus ( Acetobacter pasteurianus ), deposited in the General Microbiology Center of China Culture Collection of Microorganisms (CGMCC) on March 21, 2025, with the deposit number CGMCC No. 33924, and the deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0007] The Acetobacter pasteurianus CL247 of the present invention has the characteristics of producing acid in 72 hours of ≥49 g / L and being tolerant to 7% acetic acid and 18% ethanol.

[0008] In a second aspect of the present invention, a microbial mixture is provided, characterized by comprising at least the strain of the present invention. The microbial mixture may comprise other ethanol-resistant and acetic acid-resistant strains and microorganisms that coexist with the strain of the present invention.

[0009] In a third aspect of the present invention, a product comprising the strain of the present invention or the microbial mixture of the present invention is provided. The product may be a bacterial agent, a bacterial fertilizer or a composition.

[0010] Preferably, the product includes a carrier; the carrier may be a solid carrier or a liquid carrier. The solid carrier or liquid carrier is a conventional carrier material, wherein the solid carrier may be selected from clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica, corn flour, soybean flour, polyvinyl alcohol and / or polyglycol; the liquid carrier may be vegetable oil, mineral oil or water.

[0011] Preferably, the product is in powder form, solution form, suspension form, tablet form, granule form or other forms.

[0012] Further preferably, the powder form may be a freeze-dried powder form; and the tablet form may be a tablet form containing the freeze-dried powder and optional nutrients (such as vitamins, mineral salts, etc.).

[0013] The active ingredient of the product is Acetobacter pasteurianus, which can be present in the form of cultured living cells, fermentation broth of living cells, filtrate of cell culture or a mixture of cells and filtrate.

[0014] In a fourth aspect of the present invention, a fermentation broth containing Acetobacter pasteurianus is provided.

[0015] In a fifth aspect of the present invention, a fermentation method of Acetobacter pasteurianum is provided, which comprises inoculating the Acetobacter pasteurianum into a fermentation medium and performing fermentation culture.

[0016] In a sixth aspect of the present invention, there is provided a use of the strain of the present invention, the microbial mixture of the present invention, or the product of the present invention comprising the strain or the microbial mixture in the production of fruit vinegar.

[0017] In a seventh aspect of the present invention, a method for producing fruit vinegar is provided, comprising the step of fermenting the strain of the present invention, the microbial mixture of the present invention, or the product of the present invention comprising the strain or the microbial mixture.

[0018] In one or some embodiments of the present invention, the fermentation includes liquid submerged fermentation, and the acid production rate is ≥1.51 g / L / h, which is suitable for industrial continuous production. Compared with the related art known to the inventors, one of the technical solutions of the present invention has the following beneficial effects: The acetic acid bacteria strain CL247 of the present invention has achieved breakthroughs in comprehensive performance in the following aspects through natural selection and directed evolution: (1) Industrialized acid production efficiency: 49.2 g / L acid production in 72 hours, 1.51 g / L / h acid production rate in liquid deep fermentation, suitable for continuous fermentation needs.

[0019] (2) Dual tolerance synergy: It has 7% acetic acid tolerance and 18% ethanol tolerance, etc., supporting complex fermentation environments.

[0020] (3) Cost advantage: Reduce the frequency of feeding and the cost of raw material dilution, and improve equipment utilization.

[0021] This technical solution provides an efficient, tolerant and stable strain solution for fruit vinegar production, promoting the industry's upgrade from "inefficient and intermittent" to "efficient and continuous", and has significant technological innovation and economic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The accompanying drawings, which constitute a part of the specification of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0023] Figure 1 : Morphological characteristics of strains.

[0024] Figure 2 : Phylogenetic tree of CL247, Cp9dv1, and Cp9dv46 strains.

[0025] Figure 3 : Screening of high-acid-producing acetic acid bacteria strains and comparison of acetic acid production ability, 1: CP9dV46; 2: CL247; 3: blank; 4: CP9dV1.

[0026] Figure 4 : Diameters of transparent zones of different strain numbers.

[0027] Figure 5 : Comparison of acid production ability of different acetic acid bacteria. DETAILED DESCRIPTION

[0028] It should be noted that the following detailed descriptions are exemplary and intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs.

[0029] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments according to the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is intended to include the plural form. In addition, it should be understood that when the terms "comprise" and / or "include" are used in this specification, they indicate the presence of features, steps, operations and / or combinations thereof.

[0030] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0031] Example 1: Screening of highly ethanol-resistant and highly acid-producing acetic acid bacteria By consulting literature and relevant information, we selected the culture medium of commonly used acetic acid bacteria, screened a series of fungi and bacteria from Shanxi aged vinegar samples, and preliminarily identified the microorganisms through gene sequencing, BLAST comparison, and phylogenetic analysis. We obtained three acetic acid bacteria strains: Acetobacter pasteurianus CP9dV1 ( Acetobacter pasteurianus CP9dV1), Acetobacter pasteurianus CP9dV46 ( Acetobacter pasteurianus CP9dV46) and Acetobacter pasteurianus CL247 ( Acetobacter pasteurianus CL247). The formula of acetic acid bacteria growth medium is shown in Table 1.

[0032] (1) Isolation and culture of Acetobacter pasteurianus ① Dilution: Let the treated sample stand for 10 minutes, draw 1mL of sample supernatant and add it into a sterilized small test tube containing 9mL of distilled water. The concentration of the soil solution is 10 -2 According to this method, the soil concentration is diluted to 10 -3 , 10 -4 , 10 -5 .

[0033] ② Inoculation: Take several plates poured above, draw 100 μl of the sample supernatant into each plate with a pipette, and spread it with alcohol using a sterilized triangular spreader (the dilution concentration is 10 -3 , 10 -4 , 10 -5 , two plates were spread for each sample on acetic acid bacteria growth medium).

[0034] ③ Incubation: After inoculation, place the culture dish in a 30°C incubator for 3-5 days. Observe the acetic acid bacteria colonies regularly and record them. (The acetic acid produced by acetic acid bacteria decomposes calcium carbonate in the growth medium, resulting in a clearing zone.)

[0035] ④ Purification: Observe the morphology (size, shape and color) of the colonies on the separation culture medium at any time, pick the culture medium of corresponding concentration with an inoculation loop, dilute the sample on the plate by the plate streak method and number it in order to obtain single colonies.

[0036] ⑤Observation of morphological characteristics and preliminary identification of acetic acid bacteria strains: Use an inoculation loop to inoculate the purified strain onto the prepared acetic acid bacteria growth medium, place it in a 30℃ constant temperature incubator and culture it upside down for 3-5 days, observe its morphological characteristics under a microscope, and make a record, such as Figure 1 shown.

[0037] Table 1 Isolation and purification medium formula

[0038] Example 2: Molecular identification of Acetobacter pasteurianus Gene fragments were extracted and PCR amplified from the selected strains in a 50-ml reaction system: 1.0 μL template DNA, 2 μL upstream primer 27F, 2 μL lower primer 1492R, 10 μL 5x PCR Buffer (mg²+), 4 μL DNTP (2.5 μM each), 1 μL TagDNA Polymerase (5 U / μL), and ultrapure water was added to 50 μL. The 16S rDNA universal forward primer (SEQ ID NO: 1) was 5'-AGAGTTTGATCCTGGCTCA-3', and the 16S rDNA universal reverse primer (SEQ ID NO: 3) was 5'-GGTTACCTTGTTACGACTT-3'. The PCR reaction conditions were: 94°C pre-denaturation for 5 minutes, 94°C denaturation for 30 seconds, 50°C annealing for 30 seconds, and 72°C extension for 100 seconds. After 30 cycles, the reaction was extended at 72°C for 5 minutes and stored at 4°C. 1.0% agarose gel electrophoresis was used to check whether the desired fragment was amplified. The PCR products were then sent to the company for sequencing. BLAST alignment and phylogenetic analysis were also performed.

[0039] The gene sequence of Acetobacter pasteurianus Cp9dv1 is as follows, as shown in SEQ ID NO: 3: CCCCGGGCGGCTGCTTACCATGCAAGTCGCACGAAGGTTTCGGCCTTAGTGGCGGACGGGTGAGTAACGCGAAGGATCTATCCATGGGTGGGGGATAACACTGGGAAACTGGTGCTAATACCGCATGACACCTGAGGGTCAAAGGCGCAAAACGCCTGTGGAGGAGCCTGCGTTTGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGATGATCAATAGCTGGTTTGAGAGGATGATCAGCCACACTGGGACTGAGACACGGCCCACACTAATACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGGGCAACCCTGATCCAGCAATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTCGACGGGGACGATGATGACGGTACCCGTAGAAGAAGCCCCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGGGCTAGCGTTGCTCGGAATGACTGGGCGTAAAGGGCGTGTAGGCGGTTTGTACAGTCAGATGTGAAATCCCCGGGCTTAACCTGGGAGCTGCATTTGATACGTGCAGACTAGAGTGTGAGAGAGGGTTGTGGAATTCCCAGTGTAGAGGTGAAATTCGTAGATATTGGGAAGAACACCGGTGGCGAAGGCGGCAGCCTGGCTCATTACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAAGGATTAGATACTCTGGTAGTCCACGCTAGTCACGATGTATGCTAGATGTTGGGCGACTAATCATTCACTGC The gene sequence of Acetobacter pasteurianus Cp9dv46 is as follows, as shown in SEQ ID NO: 4: CGCTAGGCTGCTTACCATGCAAGTCGCACGAAGGTTTCGGCCTTAGTGGCGGACGGGTGAGTAACAGCGTAAGTATCTATCCATGGGTGGGGGATAACACTGGGAAACTGGTGCTAATACCGCATGACACCTGAGGGTCAAAGGCGCAAGACGCCTGTGGAGGAGCCTGCGTTTGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGATGATCAATAGCTGGTTTGAGAGGATGATCAGCCACACTGGGACTGAGACACGGCCCAGACTCATACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGGGCAACCCTGATCCAGCAATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTCGACGGGGACGATGATGACGGTACCCGTAGAAGAAGCCCCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGGGCTAGCGTTGCTCGGAATGACTGGGCGTAAAGGGCGTGTAGGCGGTTTGTACAGTCAGATGTGAAATCCCCGGGCTTAACCTGGGAGCTGCATTTGATACGTGCAGACTAGAGTGTGAGAGAGGGTTGTGGAATTCCCAGTGTAGAGGTGAAATTCGTAGATATTGGGAAGAACACCGGTGGCGAAGGCGGCAGCCTGGCTCATTACTGAGCTGAGGCGCGAAAGCGTGGGGAGCAAACAGATTAATACCCTGGTAGGTCACCTGTAACAATGTGTGCTACATGTTGGTGACTAATCATCAGG The gene sequence of Acetobacter pasteurianus CL247 is as follows, as shown in SEQ ID NO: 5: TGGGCGCATGCTTACACATGCAAGTCGCACGAAGGTTTCGGCCTTAGTGGCGGACGGGTGAGTAACGCGTAGGTATCTATCCATGGGTGGGGGATAACACTGGGAAACTGGTGCTAATACCGCATGACACCTGAGGGTCAAAGGCGCAAGTCGCCTGTGGAGGAGCCTGCGTTTGATTAGCTAGTTGGTGGGGTAAAGGCCTACCAAGGCGATGATCAATAGCTGGTTTGAGAGGATGATCAGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGGGCAACCCTGATCCAGCAATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTCGACGGGGACGATGATGACGGTACCCGTAGAAGAAGCCCCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGGGCTAGCGTTGCTCGGAATGACTGGGCGTAAAGGGCGTGTAGGCGGTTTGTACAGTCAGATGTGAAATCCCCGGGCTTAACCTGGGAGCTGCATTTGATACGTGCAGACTAGAGTGTGAGAGAGGGTTGTGGAATTCCCAGTGTAGAGGTGAAATTCGTAGATATTGGGAAGAACACCGGTGGCGAAGGCGGCAACCTGGCTCATTACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCTGTAAACGATGTGTGCTAGATGTTGGGTGACTTAGTCATTCAGTGTCGCAGTTAACGCGTTAAGCACACCGCCTGGGGAGTACGGCCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGCAGAACCTTACCAGGGCTTGAATGTAGAGGCTGCAAGCAGAGATGTTTGTTTCCCGCAAGGGACCTCTAACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTT。 Example 3: Comparison of Acid Production Capacity of Acetobacter pasteurianus The acetic acid bacteria strains of Example 2 were subjected to the Oxford cup test to preliminarily compare their acid production capabilities by the size of the transparent zone. The results are as follows: Figure 2 and Figure 3 As shown, the results showed that Acetobacterium pasteurianum CL247 had a strong ability to produce acid; This was further validated through an acid-base titration experiment: A loopful of each selected strain was inoculated into a 50ml Erlenmeyer flask containing 20ml of acid production test medium. After incubation in a constant temperature shaking incubator at 30°C and 120 rpm for 3 days, 1ml of the fermentation broth was added to a 50ml Erlenmeyer flask containing 9ml of deionized water. Three to five drops of phenolphthalein indicator were added, and the solution was titrated with 0.1mol / L NaOH solution until a light pink color was obtained. Three replicates were prepared for each treatment, and the average values ​​were compared. The volume of NaOH solution consumed by each strain was recorded. The strain that consumed the most NaOH solution was considered the highest acid producer.

[0040] Under the same fermentation conditions, the total acid content (in terms of acetic acid) was determined. Acetobacter pasteurianus CP9dV1 ( Acetobacter pasteurianus CP9dV1), Acetobacter pasteurianus CP9dV46 ( Acetobacter pasteurianus CP9dV46) and Acetobacter pasteurianus CL247 ( Acetobacter pasteurianus CL247) were 38.1 g / L, 33.8 g / L, and 49.2 g / L, respectively. The results further showed that Acetobacter pasteurianus CL247 had a strong acid production capacity. Figure 4 shown.

[0041] Example 4: Comparison of Acetate Tolerance of Acetobacter pasteurianus Acetobacter pasteurianus CL247 was inoculated at a 5% inoculum into a 100 mL Erlenmeyer flask containing 50 mL of basal liquid medium and incubated in a constant temperature shaking incubator at 30°C and 120 rpm for 2 days to obtain seed liquid. The seed liquid was then transferred at a 10% inoculum into acid production test medium containing 2%, 3%, 4%, 5%, 6%, and 7% acetic acid, all containing 3% anhydrous ethanol. Three replicates were prepared for each treatment and incubated in a constant temperature shaking incubator at 30°C and 150 rpm for 24 and 72 hours, respectively. The samples were diluted tenfold and the absorbance (strain growth) of the fermentation broth was measured at 600 nm in a UV-visible spectrophotometer. The average values ​​were compared to determine the strain's acetic acid tolerance.

[0042] Table 2 Acetic acid resistance test of Acetobacter pasteurianus CL247

[0043] Example 5: Comparison of Ethanol Tolerance of Acetobacter pasteurianus After culturing for a certain period of time with different concentration gradient ethanol, the OD 600 Changes. Acetobacter pasteurianus CL247 was inoculated into a 100mL Erlenmeyer flask containing 50mL of basal liquid culture medium and cultured in a constant temperature shaking incubator at 30°C and 150 rpm for 2 days to obtain seed liquid. The seed liquid was then transferred at a 10% inoculum into acid production test medium containing 3%, 5%, 7%, 9%, 11%, 14%, and 18% ethanol, respectively. Three parallel samples were set up for each treatment. After cultured in a constant temperature shaking incubator at 30°C and 120 rpm for 3 days, the samples were diluted tenfold and placed in a UV-visible spectrophotometer at a wavelength of 600nm to measure the absorbance of the fermentation liquid (strain growth).

[0044] Table 3 Ethanol resistance test of Acetobacter pasteurianus CL247

[0045] Example 6 Application of Apple Cider Vinegar Fermentation (1) Raw material processing After washing the apples, remove the cores, pulp the fruit and filter to obtain clarified juice.

[0046] (2) Alcohol fermentation Inoculate with brewer's yeast, with an initial sugar content of 16%, ferment at 28°C for 5 days, and the alcohol content reaches 8%-10%.

[0047] (3) Acetic acid fermentation After sterilization, the secondary seed liquid of Acetobacter pasteurianus CL247 (inoculation volume 8%) was added. The ventilation and stirring parameters of the liquid submerged fermentation in a 10L mechanical ventilation fermenter were as follows: the initial ventilation volume was 0.07m 3 / (m 3 min), mid-term 0.1-0.12m 3 / (m 3 min), 0.08m in the later period 3 / (m 3 ·min), stirring rate 150r / min, 30℃, culture and fermentation for 4 days, total acid 53.2g / L.

[0048] Through the above implementation methods, the strain CL247 of the present invention demonstrates the technical advantages of high efficiency, tolerance and stability in fruit vinegar production, promoting the industry to upgrade to efficient continuous production.

[0049] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. A strain of Acetobacter pasteurianus CL247, characterized in that: The strain was named Acetobacter pasteurianus ( Acetobacter pasteurianus ), deposited in the General Microbiology Center of China Culture Collection of Microorganisms (CGMCC) on March 21, 2025, with the deposit number CGMCC No. 33924, and the deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

2. A microbial mixture characterized by: The microbial mixture comprises at least the strain according to claim 1, and the microbial mixture comprises other ethanol-resistant and acetic acid-resistant strains and microorganisms that coexist with the strain according to claim 1.

3. A product comprising the strain according to claim 1 or the microbial mixture according to claim 2. A fermentation broth comprising the strain according to claim 1 .

5. A fermentation method of Acetobacter pasteurianus according to claim 1, characterized in that: The Acetobacter pasteurianus is inoculated into a fermentation medium for fermentation culture.

6. Use of the strain according to claim 1, the microbial mixture according to claim 2, or the product according to claim 3 in producing fruit vinegar.

7. A method for producing fruit vinegar, characterized in that: The method comprises the step of fermenting the strain according to claim 1, the microbial mixture according to claim 2 or the product according to claim 3.

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