Acetobacter breviscapus GJ1-4, microbial agent thereof and application of acetobacter breviscapus GJ1-4 in promotion of acidification of kitchen waste and biogas production

Abstract

The invention relates to acetobacter breviscapus GJ1-4, a microbial agent of the acetobacter breviscapus GJ1-4 and application of the acetobacter breviscapus GJ1-4 in promoting acidification of kitchen waste and biogas production, and belongs to the technical field of microorganisms The Acetobacter cereus GJ1-4 is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No.32450, the preservation date is October 31, 2024, and the address of the preservation institution is No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The invention further provides application of the acetobacter breviscapus GJ1-4 in promotion of acidification and biogas production of kitchen waste. The acetobacter breviscapus GJ1-4 provided by the invention can efficiently grow in a pH range of 4.0-9.0, has the characteristics of efficient acid production, high ammonia nitrogen resistance and good culture, and can effectively promote acidification of kitchen waste and production of biogas.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, specifically to a strain of Acetobacter oryzae GJ1-4 and its inoculant, and its application in promoting the acidification and biogas production of kitchen waste. Background Technology

[0002] With the acceleration of urbanization, the efficient treatment and resource utilization of food waste has become an urgent environmental problem. Anaerobic digestion technology is considered an ideal approach because it can produce clean energy, methane. However, food waste has a complex composition, and its anaerobic digestion process, especially the critical acid production stage, often faces the challenge of drastic fluctuations in environmental conditions. This results in slow system start-up, unstable operation, and unsatisfactory methane yield, severely restricting the widespread application of this technology.

[0003] The acidification stage, serving as the crucial link between hydrolysis and methanogenesis, is critically important for its efficiency and product orientation. An ideal acidification stage not only requires the rapid production of large amounts of volatile acids (VFAs) but also the targeted generation of high-quality substrates, such as acetic acid, readily available to methanogens, thereby efficiently driving subsequent processes. However, during the degradation of food waste, nitrogenous substances like proteins decompose, releasing high concentrations of ammonia nitrogen, naturally raising the system's pH. These inherent environmental pressures (high ammonia nitrogen, high pH) severely inhibit traditional acid-producing microbial communities, resulting in incomplete acidification or undesirable products (such as propionic acid accumulation), ultimately limiting the overall methanogenesis efficiency of the system.

[0004] Biofortification strategies have been extensively studied to optimize the acid production stage. However, commonly used fortification strains (such as certain lactic acid bacteria or traditional Bacillus) have significant drawbacks: they are generally poorly adapted to environmental stresses, especially the inherent high ammonia nitrogen and high pH conditions within the food waste digestion system. Under these harsh conditions, these strains are prone to inactivation, unable to stably colonize and function, resulting in insignificant and unreliable fortification effects. In other words, current technologies lack a "super acid-producing agent" that can maintain high activity throughout the anaerobic digestion environment of food waste.

[0005] Therefore, developing and applying a novel functional strain that combines high acid production capacity with excellent environmental tolerance has become the key to breaking through the current technological bottlenecks. Summary of the Invention

[0006] To address the technical problems of increased pH and incomplete acidification in fermentation broth caused by ammonia nitrogen inhibition during existing food waste treatment processes, this invention provides a strain of Acetobacter oryzae GJ1-4, its inoculum, and its application in promoting acidification and biogas production from food waste, thereby solving the aforementioned problems.

[0007] The technical solution of this invention is as follows: In a first aspect, the present invention provides a strain of Acetobacter oryzae GJ1-4, wherein the Acetobacter oryzae (Acetobacter oryzoeni GJ1-4 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 32450, on October 31, 2024. The address of the depository is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0008] Furthermore, the 16S rDNA sequence of the *Acetobacter oryzae* GJ1-4 is shown in SEQ ID NO.1.

[0009] Secondly, the present invention provides a bacterial agent containing Acetobacter oryzae GJ1-4, wherein the bacterial agent is a liquid bacterial agent.

[0010] Furthermore, the liquid bacterial agent contains ≥5 × 10⁻⁴ viable bacteria of *Acetobacter oryzae* GJ1-4. 9 CFU / mL.

[0011] A method for preparing the above-mentioned liquid bacterial agent includes the following steps: The LB culture medium of Acetobacter oryzae GJ1-4 was inoculated into the fermentation medium at a temperature of 37±1℃, a rotation speed of 200 rpm, and an aeration ratio of 1:1. Fermentation was stopped when the OD value no longer increased, and liquid inoculum was obtained.

[0012] Furthermore, the fermentation medium is composed of: 25 g / L liquid molasses, 10 g / L corn steep liquor powder, 0.3 g / L peptone, 2.5 g / L corn starch, 1 g / L dipotassium hydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 0.5 g / L calcium carbonate, and 0.12 g / L manganese sulfate monohydrate, diluted to a final volume of 850 L, sterilized at 121 °C for 20 min, and then diluted to a final volume of 1000 L.

[0013] Thirdly, the present invention provides an application of Acetobacter oryzae GJ1-4 in promoting acidification and biogas production from kitchen waste.

[0014] Furthermore, during the acidification stage of the food waste material, a microbial agent containing Acetobacter oryzae GJ1-4 is added; the inoculation amount of the microbial agent is 1%~2% (v / w) of the total weight of the food waste material.

[0015] Furthermore, a microbial agent containing Acetobacter oryzae GJ1-4 can be added at the start of fermentation, or when the pH rises above 7.5 during the fermentation of kitchen waste materials, a microbial agent containing Acetobacter oryzae GJ1-4 can be added.

[0016] The beneficial effects of this invention are as follows: This invention is the first to propose using *Acetobacter oryzae* GJ1-4 as an enhancer for the acid production stage of anaerobic digestion of food waste. Compared with existing technologies, the *Acetobacter oryzae* GJ1-4 provided by this invention specifically addresses the shortcoming of low environmental tolerance in existing enhancer strains. A performance comparison of *Acetobacter oryzae* GJ1-4 provided by this invention with traditional strains shows that the activity of existing strains is severely inhibited under high ammonia nitrogen (typically >1500 mg / L) and the resulting high pH (e.g., pH=8.0~8.5) conditions. However, *Acetobacter oryzae* GJ1-4 exhibits superior tolerance, maintaining vigorous metabolic activity and continuously and efficiently producing acid even under these conditions that inactivate traditional strains. This means that during the period when ammonia nitrogen inhibition is most likely to occur in food waste digestion, *Acetobacter oryzae* GJ1-4 not only survives but also functions effectively, ensuring the stability and continuity of the acid production process. Acetobacter oryzae GJ1-4 also possesses low-oxygen tolerance, enabling it to better adapt to the trace oxygen environment in the digestive system and quickly occupy its ecological niche, achieving rapid initiation. This operational robustness is not possessed by many strict anaerobic bacteria.

[0017] The *Acetobacter oryzae* GJ1-4 provided by this invention can withstand environmental pressure. As an acetic acid bacterium, the core function of *Acetobacter oryzae* GJ1-4 is to rapidly convert substrates into acetic acid, which provides the most direct substrate for the methanogenesis stage, fundamentally optimizing the metabolic pathway and avoiding the accumulation of inhibitory products such as propionic acid.

[0018] In summary, the *Acetobacter oryzae* GJ1-4 provided by this invention is a solution with significant performance advantages designed to address the practical pain points of anaerobic digestion of food waste. Its resistance to ammonia nitrogen, high pH, ​​and low oxygen levels is its core competitive advantage compared to existing technologies. This study aims to systematically verify that by adding *Acetobacter oryzae* GJ1-4, environmental fluctuations can be overcome, achieving highly efficient enhancement of the acid production stage, ultimately significantly improving the degradation of food waste, methane yield, and process stability, providing an innovative and reliable strategy for advancing anaerobic digestion technology. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 This is a colony diagram of Acetobacter oryzae GJ1-4 in Example 1 of the present invention.

[0021] Figure 2 This is a graph showing the pH resistance test results of Acetobacter oryzae GJ1-4 in Example 2 of the present invention.

[0022] Figure 3 This is a graph showing the ammonia nitrogen tolerance test results of *Acetobacter oryzae* GJ1-4 in Example 3 of this invention.

[0023] Figure 4 This is a graph showing the test results of the acid-producing capacity of Acetobacter oryzae GJ1-4 in Example 5 of the present invention.

[0024] Figure 5 This is a graph showing the gas production capacity test results of Acetobacter oryzae GJ1-4 in Example 6 of the present invention.

[0025] Figure 6 This is a graph showing the cumulative gas production capacity test results of *Acetobacter oryzae* GJ1-4 in Example 7 of this invention.

[0026] Figure 7 This is a graph showing the changing trends of COD and TS values ​​in the biogas slurry after continuous fermentation of *Acetobacter oryzae* GJ1-4 in Example 8 of this invention. Detailed Implementation

[0027] To enable those skilled in the art to better understand the technical solutions of this invention, the technical solutions of the embodiments of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of this invention.

[0028] Example 1 1. Sampling On August 24, 2023, anaerobic fermentation slurry was taken from the anaerobic fermentation tank of a kitchen waste treatment plant in Dezhou.

[0029] 2. Separation and screening (1) Preparation of enrichment culture medium: 10.0g glucose, 5.0g yeast extract, 5.0g peptone, 2.0g potassium dihydrogen phosphate, 0.5g magnesium sulfate, 0.1g calcium chloride, 19.1g ammonium chloride, 15.0g sodium bicarbonate, 50mL kitchen waste slurry, bring the volume up to 1000mL, pH=8.5.

[0030] (2) Preparation of screening medium: Based on the composition of enrichment medium, add 1% calcium carbonate.

[0031] (3) Enrichment culture: Take a 1L beaker, add 800mL of the enrichment culture medium prepared in step (1), and culture statically at 35℃.

[0032] (4) After multiple coatings in the screening medium, select strains with larger transparent zones to obtain single colonies, which are numbered GJ1-4.

[0033] 3. Identification Strain GJ1-4 appears as milky white, opaque, moist, smooth, irregularly shaped colonies on LB agar plates, such as... Figure 1 As shown. The sample was sent to Beijing Qingke Biotechnology Co., Ltd. for identification, and the result was *Acetobacter oryzae*. *Acetobacter oryzae* ( Acetobacter oryzoeni GJ1-4 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 32450, on October 31, 2024. The address of the depository is: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Example 2 pH resistance test of Acetobacter oryzae GJ1-4 in rice wine Seed culture medium: glucose 10g / L, yeast extract 10g / L, peptone 10g / L, pH natural. Basic fermentation medium: 1% yeast extract, 1% glucose, 6 mL anhydrous ethanol, and 94 mL distilled water.

[0034] Strain activation and seed culture preparation: The preserved Acetobacter oryzae GJ1-4 was inoculated into the seed culture medium and cultured at 30℃ and 150rpm for 24-48 hours with shaking. The culture was activated for two generations to ensure that the strain was in a vigorous growth period.

[0035] Experimental group design: Seven experimental groups were set up, with the pH adjusted to 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, and 10.0 using sodium hydroxide and hydrochloric acid. Each group had three replicates.

[0036] Fermentation Culture: Prepare the basic fermentation medium (excluding anhydrous ethanol) and dispense it into 250mL Erlenmeyer flasks (94mL of medium per flask). Autoclave (115℃, 20min). After cooling, under aseptic conditions in a laminar flow hood, add filtered sterilized anhydrous ethanol and acid / base solutions to achieve the preset pH gradient. Inoculate the activated seed culture into each experimental group's Erlenmeyer flask at a 5% inoculation rate. Incubate at 30℃ in a constant temperature shaking incubator at 150rpm for 24 hours. The growth of *Acetobacter oryzae* GJ1-4 at different pH values ​​is shown below. Figure 2 As shown.

[0037] Depend on Figure 2 It can be seen that *Acetobacter oryzae* GJ1-4 grows well in the pH range of 4.0 to 9.0, proving that *Acetobacter oryzae* GJ1-4 has significant alkali resistance.

[0038] Example 3 Ammonia nitrogen tolerance test of Acetobacter oryzae GJ1-4 Seed culture medium: glucose 10g / L, yeast extract 10g / L, peptone 10g / L, pH natural. Basic fermentation medium: anhydrous ethanol 20 g / L (sterilized separately, added before use), glucose 5 g / L, KH₂PO₄ 1 g / L, MgSO₄·7H₂O 0.5 g / L. No organic nitrogen source is added. Nitrogen source: ammonium sulfate is the sole nitrogen source.

[0039] Strain activation and seed culture preparation: The preserved Acetobacter oryzae GJ1-4 was inoculated into seed culture medium and cultured at 30°C and 150 rpm for 48 hours with shaking. The culture was continuously activated for two generations to ensure that the strain was in a vigorous growth period. Experimental group design: Seven experimental groups were set up, with ammonium chloride providing ammonia nitrogen concentration gradients of 500 mg / L, 3800 mg / L, 6800 mg / L, 9800 mg / L, 12800 mg / L, and 15800 mg / L. Each group had three replicates.

[0040] Fermentation Culture: Prepare the basic fermentation medium (excluding ethanol and nitrogen source) and dispense it into 250mL Erlenmeyer flasks (100mL medium per flask). Autoclave (115°C, 20min). After cooling, under aseptic conditions in a laminar flow hood, sequentially add filtered sterilized ethanol solution and ammonium sulfate solution of the corresponding concentration to achieve a final ethanol concentration of 20g / L and an ammonia nitrogen concentration reaching the preset gradient. Inoculate the activated seed culture into each experimental group's Erlenmeyer flask at a 5% inoculum rate. Incubate at 30°C in a constant temperature shaking incubator at 150rpm for 24 hours. The growth of the strain under different ammonia nitrogen concentrations is shown below. Figure 3 As shown.

[0041] Depend on Figure 3 It can be seen that *Acetobacter oryzae* GJ1-4 can grow well under conditions with ammonia nitrogen concentrations of 3500~9500 mg / L.

[0042] Example 4 A liquid bacterial agent containing Acetobacter oryzae GJ1-4 is prepared as follows: Five L of LB broth of *Acetobacter oryzae* GJ1-4 was inoculated into the fermentation medium. The temperature was 37±1℃, the rotation speed was 200 rpm, and the aeration ratio was 1:1. Fermentation was terminated when the OD value no longer increased, yielding a bacterial agent containing *Acetobacter oryzae* GJ1-4. Viable cell count analysis showed that the viable count of *Acetobacter oryzae* GJ1-4 was 5 × 10⁻⁶. 9 CFU / mL.

[0043] The fermentation medium consisted of: 25 g / L liquid molasses, 10 g / L corn steep liquor powder, 0.3 g / L peptone, 2.5 g / L corn starch, 1 g / L dipotassium hydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 0.5 g / L calcium carbonate, and 0.12 g / L manganese sulfate monohydrate, brought to a final volume of 850 L. The medium was then sterilized at 121 °C for 20 min, and the final volume was brought to 1000 L.

[0044] Example 5 Test of acid-producing capacity of Acetobacter oryzae GJ1-4 in rice wine Fermentation material: taken from the canteen of Anxingda (Shandong) Environmental Protection Technology Co., Ltd., after being crushed and homogenized, the total solids content was adjusted to 10% to obtain kitchen waste material.

[0045] Microbial agent: The liquid microbial agent containing Acetobacter oryzae GJ1-4 prepared in Example 4, with a viable count of 5 × 10⁻⁴ CFU / g. 9 CFU / mL.

[0046] Experimental group: Take 500g of kitchen waste material and inoculate it with 1.0% (v / w) of liquid bacterial agent containing Acetobacter oryzae GJ1-4.

[0047] Control group: 500g of kitchen waste was inoculated with the same amount of sterilized liquid microbial agent as the experimental group. The sterilized liquid microbial agent used in the control group was a liquid microbial agent containing Acetobacter oryzae GJ1-4 that had been inactivated before use.

[0048] Both groups of materials were placed in 1L anaerobic fermentation flasks and subjected to batch anaerobic fermentation in a 35±1℃ constant temperature water bath. Three parallel experimental groups were set up for each group. The curves showing the change in total volatile fatty acid concentration over time for the experimental and control groups are shown below. Figure 4 As shown.

[0049] Depend on Figure 4 It can be seen that the volatile acid content in the experimental group was 1080.805 mg / L, while that in the control group was 644.286 mg / L. Compared with the control group, the volatile acid content in the experimental group was 67.75% higher at 24 hours.

[0050] Example 6 Experiment on the enhancing effect of Acetobacter oryzae GJ1-4 in the fermentation of kitchen waste Fermentation material: taken from the canteen of Anxingda (Shandong) Environmental Protection Technology Co., Ltd., after being crushed and homogenized, the total solids content was adjusted to 10% to obtain kitchen waste material.

[0051] Microbial agent: The microbial agent containing Acetobacter oryzae GJ1-4 prepared in Example 3, with a viable count of 5 × 10⁻⁴ CFU / g. 9 CFU / mL.

[0052] Sludge: Biochemical sludge taken from the centrifuge dewatering machine of a kitchen waste treatment plant in Dezhou.

[0053] Experimental Group 1: Take 500g of kitchen waste material, inoculate with 0.5% (v / w) of a bacterial agent containing Acetobacter oryzae GJ1-4, and inoculate with 10% sludge.

[0054] Experimental Group 2: Take 500g of kitchen waste material, inoculate with 1.0% (v / w) of a bacterial agent containing Acetobacter oryzae GJ1-4, and inoculate with 10% sludge.

[0055] Control group: 500g of kitchen waste was inoculated with the same amount of sterilized liquid microbial agent as the experimental group, along with 10% sludge. The sterilized liquid microbial agent used in the control group was a liquid microbial agent containing Acetobacter oryzae GJ1-4 that had been inactivated before use.

[0056] Experimental method: The three groups of materials were placed in 1L anaerobic fermentation bottles and subjected to batch anaerobic fermentation in a constant temperature water bath at 35±1℃. Each group was set up in triplicate.

[0057] Monitoring and Results: Methane production was measured daily using the water displacement method (using a saturated sodium chloride acid solution) until production ceased. Experimental results are as follows: Figure 5 As shown.

[0058] Depend on Figure 5 The results show that the gas production of experimental groups 1 and 2 increased by 27.41% and 42.36% respectively compared with the control group. The gas production per unit of total gas (TS) was 507.1 mL / g and 566.61 mL / g for experimental groups 1 and 2 respectively, compared with 398 mL / g for the control group. The gas production per unit of total gas (VS) was 560.22 mL / g and 525.96 mL / g for experimental groups 1 and 2 respectively, compared with 439.7 mL / g for the control group.

[0059] The definitions of unit TS gas production and unit VS gas production are as follows: Gas production per unit TS: The volume of target gas produced after complete conversion of a unit mass of total solids (TS) fermentation substrate.

[0060] Unit VS gas production: The volume of target gas produced after complete conversion of a unit mass of volatile solids (VS) fermentation substrate.

[0061] Example 7 Experiment on the enhancing effect of Acetobacter oryzae GJ1-4 in continuous anaerobic digestion of kitchen waste Microbial agent: The microbial agent containing Acetobacter oryzae GJ1-4 prepared in Example 3, with a viable count of 5 × 10⁻⁴ CFU / g. 9 CFU / mL.

[0062] Food waste: taken from the residue after centrifugation at a food waste treatment plant in Dezhou.

[0063] Experimental Method: In a 100L vertical cone-bottom anaerobic fermentation system, the experimental temperature was 35℃, the effective volume was 80L, and 1.5kg of kitchen waste residue was fed daily. After stable operation, the ammonia nitrogen concentration in the tank was measured to be 7000mg / L, and the pH was approximately 8.3. After 18 days of stable operation, 1% of *Acetobacter oryzae* GJ1-4 inoculant was added. The system operated for 36 days, and relevant parameters in the anaerobic tank were monitored daily. Biogas production was recorded by a biogas flow meter. Daily biogas production monitoring results are as follows: Figure 6 As shown; the daily monitoring results for TS (Total Solids) and COD (Chemical Oxygen Demand) are as follows. Figure 7 As shown.

[0064] Depend on Figure 6 The results show that before adding the microbial agent, the total biogas production in the first 18 days was 2984.18 L. With the feed rate remaining constant, after adding the microbial agent, the total biogas production in the first 18 days was 4035.94 L, representing a 35.24% increase in total biogas production. Meanwhile, from... Figure 7 The results show that the COD and TS values ​​in the biogas slurry decreased slowly after the addition of the microbial inoculum. The COD value decreased from an average of 53281.67 mg / L before the addition of the microbial inoculum to 48336.67 mg / L after the addition of the microbial inoculum, a decrease of 9.28%. The TS value in the biogas slurry decreased from an average of 11.60% before the addition of the microbial inoculum to 10.81% after the addition of the microbial inoculum, a decrease of 6.87%. This effectively promoted the degradation of kitchen waste residue and improved the biogas production rate.

[0065] In summary, the above experiments demonstrate that the *Acetobacter oryzae* GJ1-4 of the present invention has the following advantages: (1) High pH tolerance: Acetobacter oryzae GJ1-4 can grow efficiently in the pH range of 4.0 to 9.0.

[0066] (2) High efficiency in acid production: In a culture medium with kitchen waste hydrolysate as substrate, Acetobacter glutenophilus GJ1-4 can rapidly metabolize and produce volatile fatty acids such as acetic acid. After 24 hours of culture, the volatile acid content of the experimental group was 67.75% higher than that of the control group.

[0067] (3) Tolerance to high ammonia nitrogen: Acetobacter oryzae GJ1-4 can grow under conditions of ammonia nitrogen concentration of 3500~9500 mg / L, showing excellent ammonia nitrogen tolerance.

[0068] (4) Good culture characteristics: Acetobacter oryzae GJ1-4 can be cultured aerobically, but it is not demanding in terms of dissolved oxygen. It can grow well and perform acid production under microaerobic to anaerobic conditions. This characteristic makes it easy to scale up its culture under conventional laboratory conditions, showing good potential for industrial application.

[0069] (5) Promote the degradation of kitchen waste: Acetobacter oryzae GJ1-4 can effectively promote the degradation of kitchen waste and increase biogas production.

[0070] Although the present invention has been described in detail with reference to the accompanying drawings and preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made to the embodiments of the present invention by those skilled in the art without departing from the spirit and essence of the invention, and such modifications or substitutions should all be within the scope of the present invention. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should also be covered within the protection scope of the present invention.

Claims

1. A strain of Acetobacter oryzae GJ1-4, characterized in that, The rice wine acetic acid bacteria ( Acetobacter oryzoeni GJ1-4 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 32450, on October 31, 2024. The address of the depository is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

2. The *Acetobacter glycinobacter* GJ1-4 as described in claim 1, characterized in that, The 16S rDNA sequence of the *Acetobacter oryzae* GJ1-4 is shown in SEQ ID NO.

1.

3. A bacterial agent containing *Acetobacter oryzae* GJ1-4 as described in claim 1, characterized in that, The bacterial agent is a liquid bacterial agent.

4. The microbial agent as described in claim 3, characterized in that, The liquid bacterial agent contains ≥5 × 10⁻⁶ viable bacteria of *Acetobacter oryzae* GJ1-4. 9 CFU / mL.

5. The microbial agent as described in claim 3, characterized in that, The preparation method of the liquid bacterial agent includes the following steps: The LB culture medium of Acetobacter oryzae GJ1-4 was inoculated into the fermentation medium at a temperature of 37±1℃, a rotation speed of 200 rpm, and an aeration ratio of 1:

1. Fermentation was stopped when the OD value no longer increased, and liquid inoculum was obtained.

6. The microbial agent as described in claim 5, characterized in that, The fermentation medium consisted of: 25 g / L liquid molasses, 10 g / L corn steep liquor powder, 0.3 g / L peptone, 2.5 g / L corn starch, 1 g / L dipotassium hydrogen phosphate, 0.5 g / L magnesium sulfate heptahydrate, 0.5 g / L calcium carbonate, and 0.12 g / L manganese sulfate monohydrate, brought to a final volume of 850 L. The medium was then sterilized at 121 °C for 20 min, and the final volume was brought to 1000 L.

7. The application of Acetobacter oryzae GJ1-4 as described in claim 1 in promoting acidification and biogas production from kitchen waste.

8. The application as described in claim 7, characterized in that, During the acidification stage of kitchen waste, add a microbial agent containing Acetobacter oryzae GJ1-4.

9. The application as described in claim 8, characterized in that, The inoculation amount of the microbial agent is 1% to 2% of the total weight of the kitchen waste.

10. The application as described in claim 8, characterized in that, Add a microbial agent containing Acetobacter oryzae GJ1-4 when fermentation starts, or add a microbial agent containing Acetobacter oryzae GJ1-4 when the pH rises above 7.5 during the fermentation of kitchen waste.

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