A weissella fusion and application thereof

Abstract

The application provides a kind of fusion weissella and its application, fusion weissella is preserved in China typical culture preservation center on January 29, 2024, and the preservation number is CCTCC NO:M2024244, and the classification designation is Weissella confusa R2210.The fusion weissella provided by the application can be used for preparing fermented chili, and the fusion weissella can increase the content of flavoring substances of fermented chili products, speed up the acid production speed of fermented chili, and obtain fresh red chili with uniform color, bright and lustrous, fermented aroma, coordinated smell, rich flavor, pure and proper spiciness, soft taste, salty and light suitable, delicious taste.The process of preparing fermented chili is simple and easy to operate, and is suitable for large-scale industrial production.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, and in particular to a fusion strain of Weissella and its applications. Background Technology

[0002] Pickled chili, also known as pickled pepper, is a specialty condiment characterized by its bright red color, rich aroma, and spicy flavor. It has a unique combination of sour, spicy, sweet, savory, and salty tastes, making it suitable for various cooking methods, such as making fried rice, shredded pork with garlic sauce, and pickled chili ribbonfish.

[0003] Currently, traditional methods of preparing pickled chili peppers involve adding salt, ginger, and garlic to fresh chili peppers for natural fermentation. However, when applied to industrial production, it has been found that the taste and flavor of the pickled chili peppers are unsatisfactory, resulting in low consumer acceptance, slow fermentation speed, and poor economic benefits. This invention isolates a lactic acid bacteria strain from naturally fermented pickled chili peppers. Using this strain to prepare fermented pickled chili peppers yields peppers with a uniform color, bright and glossy appearance, a fragrant and harmonious fermented aroma, rich flavor, pure spiciness, mild taste, and a suitable level of saltiness, resulting in a delicious flavor that meets consumer demands. Furthermore, the production process is simple and easy to operate, making it suitable for large-scale industrial production. Summary of the Invention

[0004] The purpose of this invention is to provide a fused Weissella strain and its application. The fused Weissella strain provided by this invention can improve the flavor of pickled chili peppers, accelerate the fermentation process, meet consumer demand, and enhance enterprise competitiveness.

[0005] The technical solution of this invention is: a fusion strain of *Westernella*, which was deposited at the China Center for Type Culture Collection on January 29, 2024, with accession number CCTCC NO: M 2024244, and classified as... Weissella confusa R2210.

[0006] The aforementioned fused Weiss bacteria were isolated from naturally fermented pickled chili peppers.

[0007] The aforementioned application of *Westernella fusionis* in fermented chili pepper products.

[0008] The fermentation strains used in the fermentation process of the aforementioned pickled chili products include the aforementioned Fusion Weissella.

[0009] The fermentation strain used in the fermentation process of the aforementioned pickled chili products is the aforementioned Fusion Weissella.

[0010] The aforementioned fusion of Weissella bacteria can increase the content of flavor compounds in fermented chili products and accelerate the acid production rate of fermented chili.

[0011] The preparation method of the aforementioned fermented chili pepper product is carried out according to the following steps: (1) Raw material preparation: Wash the fresh red chili peppers, remove the stems from the chili peppers, cut them into small pieces of 2-8mm, add 4-8% salt, 10-14% ginger, 10-14% garlic, 2-4% soybeans, and 1.5-3% white wine, mix well to obtain product A, and set aside. (2) Cleaning and disinfection of fermentation tank: Clean the fermentation tank, dry it at 30-60℃, sterilize and disinfect it with ultraviolet light, and set it aside for later use; (3) Preparation of protective agent: 4.0-6.0g of trehalose, 7.0-9.0g of skim milk powder and 3.0-5.0g of glycerin are added to 100mL of distilled water and mixed well to obtain the protective agent for later use; (4) Preparation of direct-inoculation starter culture: Take fusion Weissella ( Weissella confusa R2210) was added to a protectant and mixed thoroughly, then combined with Weissella foenum-graecum ( Weissella confusa The ratio of R2210 to protectant was 1g:4-6ml. The mixture was dispensed into plates and freeze-dried under vacuum to obtain a direct-inoculation fermentation agent. (5) Fermentation: Add 0.6-1.0% of direct-inoculation fermentation agent to product A, mix well, put it into a cleaned and disinfected fermentation tank, and ferment at 20-40℃ for 20-35 days. The fermentation environment is an oil-free environment, and the fermented chili product is obtained.

[0012] Specifically, the preparation method of the aforementioned fermented chili pepper product is carried out according to the following steps: (1) Raw material preparation: Wash the fresh red chili peppers, remove the stems from the chili peppers, cut them into small pieces of 3-7mm, add 5-7% salt, 11-13% ginger, 11-13% garlic, 2-3% soybeans, and 1.5-2.5% white wine, mix well to obtain product A, and set aside. (2) Cleaning and disinfection of fermentation tank: Clean the fermentation tank, dry it at 30-50℃, and sterilize and disinfect it by ultraviolet irradiation for 30-60 minutes. (3) Preparation of protective agent: 4.5-5.5g of trehalose, 7.5-8.5g of skim milk powder and 3.5-4.5g of glycerin are added to 100mL of distilled water and mixed well to obtain the protective agent for later use; (4) Preparation of direct-inoculation starter culture: Take fusion Weissella ( Weissella confusa R2210) was added to a protectant and mixed thoroughly, then combined with Weissella foenum-graecum ( Weissella confusa The ratio of R2210 to the protectant was 1g:4.5-5.5ml. The mixture was dispensed into plates and then freeze-dried under vacuum to obtain a direct-inoculation fermentation agent. (5) Fermentation: Add 0.7-85% direct-inoculation fermentation agent to product A, mix well, put it into a cleaned and disinfected fermentation tank, and ferment at 25-35℃ for 25-30 days. The fermentation environment is an oil-free environment, and the fermented chili product is obtained.

[0013] More specifically, the preparation method of the aforementioned fermented chili pepper product is carried out according to the following steps: (1) Raw material preparation: Wash the fresh red chili peppers, remove the stems from the chili peppers, cut them into small pieces of 4-6mm, add 6% salt, 12% ginger, 12% garlic, 2.5% soybeans and 2% white wine, mix well to obtain product A, and set aside. (2) Cleaning and disinfection of fermentation tank: Clean the fermentation tank, dry it at 40℃, and then sterilize and disinfect it with ultraviolet light for later use; (3) Preparation of protective agent: 5.0g of trehalose, 8.0g of skim milk powder and 4.0g of glycerin were added to 100mL of distilled water and mixed well to obtain the protective agent for later use; (4) Preparation of direct-inoculation starter culture: Take fusion Weissella ( Weissella confusa R2210) was added to a protectant and mixed thoroughly, then combined with Weissella foenum-graecum ( Weissella confusa R2210) was mixed with a protectant at a ratio of 1g:5ml, dispensed into plates, and then freeze-dried under vacuum to obtain a direct-inoculation fermentation agent. (5) Fermentation: Add 0.75% direct-inoculation fermentation agent to product A, mix well, put it into a cleaned and disinfected fermentation tank, ferment at 30℃ for 30 days, and the fermentation environment is an oil-free environment to obtain the fermented chili product.

[0014] The aforementioned *Weissella fusionis* has been deposited, and its complete deposit information is as follows: Preservation institution code: CCTCC Address: Wuhan University, Bayi Road, Wuchang District, Wuhan, Hubei Province, China Survival status: Yes Deposit date: January 29, 2024 Accession number: CCTCC NO: M2024244 Category Naming: Weissella confusa R2210 The beneficial effects of this invention are: 1. The Fusion Weissella bacillus provided by this invention ( Weissella confusa R2210 can be used to prepare fermented chili peppers. The resulting chili peppers are a uniform bright red color, bright and glossy, with a fragrant fermentation aroma, harmonious smell, rich flavor, pure spiciness, mild taste, appropriate saltiness, and delicious taste, combining sourness, spiciness, freshness, and fragrance.

[0015] 2. Fermenting pickled chili peppers with *Westernella* fusion accelerates the production of lactic acid and citric acid, thus shortening the fermentation time. It also increases the content of lactic acid, citric acid, and malic acid, achieving a lactic acid content of 10677.27 mg / kg (1.62 times that of naturally fermented peppers), a citric acid content of 1191.52 mg / kg (compared to less than 3.12 mg / kg in naturally fermented peppers), and a malic acid content of 24.70 mg / kg (1.88 times that of the control group). This results in a milder and more harmonious sour taste, contributing to improved acidity quality. 3. Compared to naturally fermented peppers, fermented pickled chili peppers inoculated with *Westernella* fusion exhibit higher levels and more diverse characteristic flavor compounds, resulting in a richer flavor. Naturally fermented pickled chili peppers showed 57 flavor compounds, while those prepared with *Westernella* fusion showed 71. (1) Among the alcohol flavor substances, the linalool content of naturally fermented linalool is 6.481 mg / kg, while the linalool content of the present invention is 56.439 mg / kg; the phenylethanol content of naturally fermented phenethyl alcohol is 0.393 mg / kg, while the phenylethanol content of the present invention is 35.196 mg / kg; the citronellol content of naturally fermented citronellol is 3.289 mg / kg, while the citronellol content of the present invention is 46.525 mg / kg; the geraniol content of naturally fermented geraniol is 29.690 mg / kg, while the geraniol content of the present invention is 121.850 mg / kg. This invention integrates Weissella to impart a rich floral and honey aroma to pickled chili peppers; (2) Among the ester flavor substances, (Z)-3,7-dimethyl-2,6-octadien-1-ol acetate, which has a citrus aroma, has a content of 27.259 mg / kg in natural fermentation, while the pickled chili peppers fermented with Weissella in this invention have a content of 138.100 mg / kg, which is 5.07 times that of natural fermentation. At the same time, 2-phenylethyl propionate, which has a honey and strawberry flavor, and ethyl nonanoate, a wine-scented ester compound with a fruity and rose-like aroma, were also detected. These were not detected in natural fermentation; (3) Among the olefin flavor substances, α-curcumene has a balsamic aroma, with a content of 10.680 mg / kg in natural fermentation, while the content of fermented chili peppers fermented with Weissella in this invention is 190.736 mg / kg; β- Sesquiclane has a fruity aroma. The content of naturally fermented sucralose is 11.300 mg / kg, while the content of sucralose fermented with Weissella in this invention is 236.464 mg / kg. (4) Among the aldehyde flavor substances, citral, which has a strong lemon aroma, has a content of 13.008 mg / kg of naturally fermented sucralose, while the content of sucralose fermented with Weissella in this invention is 82.103 mg / kg. Attached Figure Description

[0016] Figure 1 Changes in the pH value of the bacterial culture; Figure 2 Changes in the total acid content of the bacterial culture; Figure 3 Colony morphology and microscopic examination results of R10 (A: Colony morphology; B: Microscopic examination results); Figure 4 : Phylogenetic tree of R10; Figure 5 Changes in pH of pickled chili peppers treated with different inoculation amounts; Figure 6 Changes in sensory scores of pickled peppers treated with different inoculation amounts; Figure 7 Changes in pH of pickled chili peppers treated with different amounts of salt; Figure 8 Changes in pH of pickled chili peppers treated with different fermentation temperatures; Figure 9 Changes in sensory scores of pickled chili peppers treated with different fermentation temperatures; Figure 10 Changes in pH of pickled chili peppers treated with different fermentation times; Figure 11 Changes in sensory scores of pickled chili peppers treated with different fermentation times. Detailed Implementation

[0017] The present invention will be further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the invention.

[0018] Example 1: Preparation method of fermented chili pepper products (1) Raw material preparation: Wash the fresh red chili peppers, remove the stems from the chili peppers, cut them into small pieces of 4-6mm, add 6% salt, 12% ginger, 12% garlic, 2.5% soybeans and 2% white wine, mix well to obtain product A, and set aside. (2) Cleaning and disinfection of fermentation tank: Clean the fermentation tank, dry it at 40℃, and sterilize and disinfect it by ultraviolet irradiation for 30-60 minutes. (3) Preparation of protective agent: 5.0g of trehalose, 8.0g of skim milk powder and 4.0g of glycerin were added to 100mL of distilled water and mixed well to obtain the protective agent for later use; (4) Preparation of direct-inoculation starter culture: Take fusion Weissella ( Weissella confusa R2210) was added to a protectant and mixed thoroughly, then combined with Weissella foenum-graecum ( Weissella confusa R2210) was mixed with a protectant at a ratio of 1g:5ml, dispensed into plates, and then freeze-dried under vacuum to obtain a direct-inoculation fermentation agent. (5) Fermentation: Add 0.75% direct-inoculation fermentation agent to product A, mix well, put it into a cleaned and disinfected fermentation tank, ferment at 30℃, the fermentation environment is an oil-free environment, ferment for 30 days, and the fermented chili product is obtained.

[0019] Example 2: Preparation method of fermented chili pepper products (1) Raw material preparation: Wash the fresh red chili peppers, remove the stems from the chili peppers, cut them into small pieces of 2-8mm, add 4% salt, 10% ginger, 10% garlic, 2% soybeans and 1.5% white wine, mix well to obtain product A, and set aside. (2) Cleaning and disinfection of fermentation tank: Clean the fermentation tank, dry it at 30℃, and sterilize and disinfect it by ultraviolet irradiation for 30-60 minutes. (3) Preparation of protective agent: 4.0g of trehalose, 7.0g of skim milk powder and 3.0g of glycerin are added to 100mL of distilled water and mixed well to obtain a protective agent for later use; (4) Preparation of direct-inoculation starter culture: Take fusion Weissella ( Weissella confusa R2210) was added to a protectant and mixed thoroughly, then combined with Weissella foenum-graecum ( Weissella confusa The ratio of R2210 to the protectant was 1g:4ml. The mixture was dispensed into plates and freeze-dried under vacuum to obtain a direct-inoculation fermentation agent. (5) Fermentation: Add 0.6% direct-inoculation fermentation agent to product A, mix well, put it into a cleaned and disinfected fermentation tank, ferment at 40℃, the fermentation environment is an oil-free environment, ferment for 20 days, and the fermented chili product is obtained.

[0020] Example 3: Preparation method of fermented chili pepper products (1) Raw material preparation: Wash the fresh red chili peppers, remove the stems from the chili peppers, cut them into small pieces of 2-8mm, add 8% salt, 14% ginger, 14% garlic, 4% soybeans and 3% white wine, mix well to obtain product A, and set aside. (2) Cleaning and disinfection of fermentation tank: Clean the fermentation tank, dry it at 60℃, and sterilize and disinfect it by ultraviolet irradiation for 30-60 minutes. (3) Preparation of protective agent: 6.0g of trehalose, 9.0g of skim milk powder and 5.0g of glycerin are added to 100mL of distilled water and mixed well to obtain a protective agent for later use; (4) Preparation of direct-inoculation starter culture: Take fusion Weissella ( Weissella confusa R2210) was added to a protectant and mixed thoroughly, then combined with Weissella foenum-graecum ( Weissella confusa The ratio of R2210 to the protectant was 1g:6ml. The mixture was dispensed into plates and freeze-dried under vacuum to obtain a direct-inoculation fermentation agent. (5) Fermentation: Add 1.0% direct-inoculation fermentation agent to product A, mix well, put it into a cleaned and disinfected fermentation tank, ferment at 20℃, the fermentation environment is an oil-free environment, ferment for 35 days, and the fermented chili product is obtained.

[0021] Example 4: Preparation method of fermented chili pepper products (1) Raw material preparation: Wash the fresh red chili peppers, remove the stems from the peppers, cut them into small pieces of 2-8mm, add 5% salt, 13% ginger, 11% garlic, 2.5% soybeans and 2.5% white wine, mix well to obtain product A, and set aside. (2) Cleaning and disinfection of fermentation tank: Clean the fermentation tank, dry it at 50℃, and sterilize and disinfect it by ultraviolet irradiation for 30-60 minutes. (3) Preparation of protective agent: 4.5g of trehalose, 8.0g of skim milk powder and 5.0g of glycerin are added to 100mL of distilled water and mixed well to obtain a protective agent for later use; (4) Preparation of direct-inoculation starter culture: Take fusion Weissella ( Weissella confusa R2210) was added to a protectant and mixed thoroughly, then combined with Weissella foenum-graecum ( Weissella confusa The ratio of R2210 to the protectant was 1g:5ml. The mixture was dispensed into plates and freeze-dried under vacuum to obtain a direct-inoculation fermentation agent. (5) Fermentation: Add 0.85% direct-inoculation fermentation agent to product A, mix well, put it into a clean and disinfected fermentation tank, ferment at 30℃, the fermentation environment is an oil-free environment, ferment for 30 days, and the fermented chili product is obtained.

[0022] This invention also conducted extensive experimental studies to demonstrate the beneficial effects of fusing Weissella oryzae in the preparation of fermented chili pepper products. The results of these experimental studies are as follows: 1 Fusion Weissella ( Weissella confusa Isolation, purification and identification of strain R2210 1.1 Experimental Materials Fresh red chili peppers: sourced from the Guiyang Huaxi Shiban planting base; soybeans, ginger, garlic, and fermentation tanks were purchased from the farmers' market; salt, liquor, raw rapeseed oil, and sugar were purchased from the supermarket.

[0023] 1.1.1 Experimental Reagents

[0024] 1.1.2 Experimental Instruments and Equipment

[0025] 1.2 Isolation and purification of lactic acid bacteria 1.2.1 Isolation and purification process of lactic acid bacteria The isolation and purification of lactic acid bacteria were performed according to GB / T 4789.35-2016. 25g of naturally fermented pickled chili peppers were weighed and placed in 225mL of sterile physiological saline. The mixture was shaken well to prepare a series of dilutions. 0.1mL of each dilution was transferred and spread onto MRS solid agar plates, incubated at 28℃ for 48-72h. Single colonies of different morphologies were picked and streaked onto MRS plates for purification until strains with consistent color, morphology, and size were obtained. The purified lactic acid bacteria were then transferred to test tube slant agar and incubated for 48-72h before being stored at 4℃ for later use. For lactic acid bacteria requiring long-term storage, the purified single colonies were cultured in liquid form, and the resulting bacterial solution was added to 30% glycerol and stored at -80℃.

[0026] 1.2.2 Results of isolation and purification of lactic acid bacteria Ten strains of lactic acid bacteria were isolated from naturally fermented pickled chili peppers and numbered R1, R2, R3, R4, R5, R6, R7, R8, R9, and R10.

[0027] 1.3 Screening of Lactic Acid Bacteria 1.3.1 Screening methods for lactic acid bacteria The purified strain was inoculated into MRS liquid medium and cultured in a constant temperature incubator at 28°C. The pH and total acid of the bacterial solution were measured, and strains with faster acid production rate and larger acid production were selected for use as fermentation strains in the later stage.

[0028] The total acid content was determined according to GB / T 12456-2021 using the acid-base indicator titration method. Based on the principle of acid-base neutralization, the acid in the bacterial culture was titrated with alkali solution, and the titration endpoint was determined using phenolphthalein as an indicator. The total acid content in the sample was calculated based on the amount of alkali solution consumed, expressed as lactic acid.

[0029] 1.3.2 Screening results of strains The pH changes of the selected strains over 48 hours after liquid culture are as follows: Figure 1 The change in total acid content is as follows Figure 2 As shown. By Figure 1 It can be seen that within 48 hours, the pH value of all 10 strains showed a decreasing trend with increasing time. The rate of pH decrease was faster within 0-12 hours, and the decreasing trend gradually slowed down after 12 hours. The pH value changes of R2 and R5 were smaller. Within 24 hours, the pH values ​​of R1, R3, R4, R8, and R10 showed larger changes, and the rate of decrease gradually decreased and tended to level off as the culture time extended. Overall, the pH values ​​of R1, R8, and R10 decreased faster, especially R10, whose pH value decreased from 6.40 to 3.86 after 48 hours of culture, a decrease of 39.7% compared with the initial pH value, indicating that R10 produced acid at a faster rate.

[0030] Depend on Figure 2 It can be seen that the total acid content increases continuously with the increase of culture time within 48 hours. The total acid content of R2, R3, and R4 changes relatively little, while the total acid content of R1, R8, and R10 increases more rapidly. pH value and total acid content are important indicators reflecting the quality of fermented products. Generally, in the early stage of fermentation, the strains mainly follow a homofermentation mode, with lactic acid content gradually increasing and pH value gradually decreasing. In the later stage of fermentation, a heterofermentation mode is dominant, with lactic acid content gradually decreasing and acetic acid content increasing. Considering the changes in pH value and total acid content, R10 was selected as the subsequent fermentation strain.

[0031] 1.4 Identification of Lactic Acid Bacteria 1.4.1 Preliminary identification of lactic acid bacteria Strain R10 was inoculated into MRS solid medium, and the morphological characteristics of the colonies, such as shape, size, color, transparency, and edge condition, were observed and recorded.

[0032] Table 1. Colony morphology of R10

[0033] like Figure 3 As shown, the colony morphology of R10 ( Figure 3 A) is round, opaque, with neat edges, 1-3 mm in size, white in color, and glossy; Gram staining and microscopic examination ( Figure 3 B) is a Gram-positive coccus. Based on Bergey's Manual of Bacteriological Identification and the Classification, Identification and Laboratory Methods of Lactic Acid Bacteria, combined with colony morphology, Gram staining and other results, the screened strains are consistent with the characteristics of lactic acid bacteria.

[0034] 1.4.2 Molecular biological identification of lactic acid bacteria Strains R10 were cultured and DNA was extracted. This DNA was amplified by PCR, followed by 16S rDNA sequencing. The sequencing results were then modified and assembled using CExpress to obtain the sequence. The sequence was then BLAST-aligned against the NCBI database to construct a phylogenetic tree. Figure 4 As shown, the isolated strain R10 is on the same branch as the standard strain Weissella confusa, and the sequence similarity is greater than 99%. Therefore, R10 was identified as Weissella confusa.

[0035] Fusion Weissella ( Weissella confusa The 16S rDNA sequence number (R2210) is as follows: 2. Direct-inoculation fusion Weissella ( Weissella confusa Preparation process of R2210 fermentation agent 2.1 Selection of Protective Agent Freeze-drying protectants, also known as freeze protectants, are substances that prevent bioactive materials from being destroyed during freeze-drying. Common freeze-drying protectants include lactose, trehalose, glucose, xylan, and glycerol. This experiment used trehalose, skim milk powder, and glycerol as freeze-drying protectants.

[0036] 2.2 Preparation of direct-inoculation fermentation agent Selected fusion Weissella ( Weissella confusa The R2210 strain was cultured and centrifuged at 5000 rpm at 25°C for 15 min. The supernatant was discarded and the bacterial cells were collected. The bacterial cells and protectant were mixed at a ratio of 1:5 (w / v, where the protectant consisted of 5.0% trehalose, 8.0% skim milk powder, and 4.0% glycerol), dispensed into plates, and then freeze-dried under vacuum to obtain the direct-inoculation fermentation broth.

[0037] 2.3 Calculation of viable cell rate Method for calculating the viability of freeze-dried cells: Take 1.0 g of freeze-dried bacterial powder and add it to 10 mL of physiological saline. Rehydrate in a 37℃ water bath for 20 min. Dilute the rehydrated fermentation agent serially with sterile water and calculate the colony count according to GB / T 4789.1-2022.

[0038] Colony forming units (CFU) per mL of sample = average colony count of three replicates at the same dilution × dilution factor × 10.

[0039] Cell viability (%) after freeze-drying = number of viable bacteria per mL after resuspending freeze-dried bacterial powder in an equal volume of physiological saline / number of viable bacteria per mL of sample before freeze-drying × 100.

[0040] Table 2. Preparation and viability results of direct-inoculation fermentation agent.

[0041] 3. Confirmation of the fermentation process for pickled chili peppers 3.1 Optimization of Fermentation Process for Pickled Chili Peppers (1) Wash the fresh red chili peppers, remove the stems, and cut them into small pieces about 5 mm in size; (2) Add salt, 10-14% of cleaned and chopped ginger, 10-14% of cleaned and chopped garlic, 2-4% of soybeans, and 1.5-3% of white wine to the chili peppers and mix well. (3) Clean the fermentation tank, dry it at 40℃, and then sterilize and disinfect it by ultraviolet irradiation for 30-60 minutes; (4) Add the direct-inoculation fermentation agent to 150g of mixed raw and auxiliary materials, mix evenly, and then put it into the disinfected fermentation tank; (5) Ferment at 30℃ to obtain fermented pickled chili products; (6) The entire process is carried out in an oil-free environment.

[0042] 3.2 Optimization of Fermentation Process The fermentation process was optimized by using the inoculum size of the direct-inoculation starter, the amount of salt added, the fermentation temperature, and the fermentation time as single factors.

[0043] 3.3 Single-factor experiment (1) Determination of the inoculum amount of direct-inoculation fermentation agent: Direct-inoculation fermentation agent was added to fresh chopped peppers at inoculum amounts of 0.25%, 0.5%, 0.75%, 1.0%, and 1.25%, respectively, with salt added at 8%. The fermentation temperature was 25℃. After 30 days of fermentation, the indicators of each group of products were measured and the sensory evaluation was carried out. Three parallel experiments were set up for each group of experiments to screen the appropriate inoculum amount.

[0044] (2) Determination of the level range of salt addition: Add 0.75% of direct-inoculation fermentation agent to fresh chopped peppers. The salt addition amounts are 2%, 4%, 6%, 8% and 10%, respectively. The fermentation temperature is 25℃. After 30 days of fermentation, the indicators of each group of products are measured and sensory evaluation is performed. Three parallel experiments are set up for each group of experiments to screen the appropriate salt addition amount.

[0045] (3) Determination of fermentation temperature: Add 0.75% of direct-inoculation fermentation agent to fresh chopped peppers, and add 8% salt. The fermentation temperature is set at 15℃, 20℃, 25℃, 30℃ and 35℃ respectively. After fermentation for 30 days, the indicators of each group of products are measured and sensory evaluation is carried out. Three parallel experiments are set up for each group of experiments to screen the appropriate fermentation temperature range.

[0046] (4) Determination of fermentation time: Add 0.75% of direct-inoculation fermentation agent to fresh chopped peppers, add 8% salt, ferment at 25℃, and set fermentation time to 15d, 20d, 25d, 30d and 35d respectively. At the corresponding fermentation time, measure the index and sensory evaluation of each group of products. Set up three parallel experiments for each group to screen the appropriate fermentation time level.

[0047] 3.4 Orthogonal Experiment to Optimize Fermentation Process Based on the single-factor experiments, the optimal fermentation conditions were determined through orthogonal experiments. The factor level table and combination table are shown in Table 3.

[0048] Table 3 Factor Level Table

[0049] 3.5 Sensory Evaluation Criteria A sensory evaluation panel of 10 people with food science backgrounds was formed. The evaluation method was as follows: 20g of fermented, mature pickled chili peppers were weighed and placed in a white container. Their color and appearance were observed under natural light, their aroma was smelled, and after rinsing their mouths with warm water, they were tasted. After each sample was evaluated, a two-minute interval was required before moving on to the next sample. The score was the sum of all scores, with a maximum score of 100. A score below 48 points resulted in the product losing its commercial value. The sensory evaluation criteria are shown in Table 4.

[0050] Table 4 Sensory Evaluation Criteria for Pickled Chili Peppers

[0051] 3.6 Results of Fermentation Process Optimization 3.6.1 Results of Single-Factor Experiment to Explore Optimal Fermentation Factors (1) Effect of direct-inoculation fermentation inoculum amount on pH and sensory score of pickled chili peppers The effect of direct-inoculation starter on the fermentation of pickled chili peppers is as follows: Figure 5 Sensory ratings can be found Figure 6 .

[0052] from Figure 5 It can be seen that the pH initially decreases and then increases with increasing inoculum amount. The lowest pH is observed at 0.5%, followed by 0.75%, then 1.25%, with the highest at 1%. Lower pH values ​​result in higher acid production. The optimal values ​​are 0.5%, 0.75%, and 1.25%.

[0053] from Figure 6 It can be seen that the inoculation amount generally showed a trend of first increasing and then decreasing. After 0.75%, the sensory evaluation of the pickled chili peppers continued to decline. Among them, the pickled chili peppers with the highest sensory score and the best quality were those with 0.75%, followed by 0.5%, while the lowest score and the worst quality were those with 1.25%. The main reason is that excessive use of direct-inoculation fermenting agents weakens the role of other microorganisms, which is not conducive to the formation of multiple flavors, resulting in an overly singular flavor. In addition, excessive use of direct-inoculation fermenting agents increases costs.

[0054] Based on the above, 0.5%, 0.75%, and 1% were selected as the experimental factors for the next step.

[0055] (2) Effect of salt addition on pH and sensory score of pickled chili peppers The effect of salt addition on the fermentation of pickled chili peppers is as follows: Figure 7 .from Figure 7The pH level shows an upward trend, with the lowest pH at 2% salt addition, followed by 4%, then 6%, and the highest at 8%. The lower the pH, the greater the acid production. High salt concentrations in fermentation can cause an imbalance between the acidic and salty tastes. Sensory scores initially rise and then fall with increasing salt content, reaching a high of 6%, followed by 8%, and a low of 10%. High salt levels inhibit the growth of lactic acid bacteria in pickled chili peppers, thus slowing down the fermentation process and worsening the flavor. Furthermore, pickled chili peppers with a maximum salt content of 10% will be very salty, affecting the taste. Excessive salt also has a certain inhibitory effect on microbial growth and acid production.

[0056] In summary, 4%, 6%, and 8% were chosen as factors for subsequent experiments.

[0057] (3) Effects of fermentation temperature on pH and sensory scores of pickled chili peppers The effect of temperature on the fermentation of pickled chili peppers, such as Figure 8 Sensory ratings can be found Figure 9 .Depend on Figure 8 It can be observed that as temperature increases, pH first decreases and then increases, but reaches its lowest point at 25℃, possibly because 25℃ is the optimal temperature for the fermentation microorganisms in pickled chili peppers. The pH is lowest at 25℃ and highest at 15℃, possibly because the temperature is not yet at the optimal range for the fermentation microorganisms. The pH is higher at 35℃, indicating that the growth and reproduction of the fermentation microorganisms are inhibited at this temperature. Temperature inhibits the growth of the fermentation microorganisms, thus affecting acid production; therefore, temperature is a crucial factor influencing fermentation.

[0058] Depend on Figure 9 It can be seen that the sensory evaluation is best at 30℃, followed by 20℃, 25℃, and 30℃, with 35℃ being the worst. Therefore, 20℃, 25℃, and 30℃ were chosen as the factors for the next stage of the experiment.

[0059] (4) Effects of fermentation time on pH and sensory scores of pickled chili peppers The effects of different fermentation times on the fermentation of pickled chili peppers are shown in the results. Figure 10 Sensory ratings can be found Figure 11 .from Figure 10 It can be seen that the pH generally decreased over time. The lowest pH was observed at 35 days, followed by 25 days, 30 days, and 20 days, with the highest pH at 15 days. The rate of pH decrease slowed after 30 days, indicating that the fermentation of the pickled chili peppers was affected, and the growth of lactic acid bacteria was inhibited by acidity. Figure 11 It can be seen that the sensory scores gradually decreased after 20 days, with the best flavor of the pickled chili peppers at 25 days, followed by 20 days and 30 days. Although the pH was very low at 35 days of fermentation, the flavor of the pickled chili peppers was very poor at this time, and the color turned black, making them inedible. Therefore, considering both pH and sensory scores, 20 days, 25 days, and 30 days were selected as factors for subsequent fermentation experiments.

[0060] 3.6.2 Results of orthogonal experiment to optimize fermentation process Based on single-factor experiments on four factors—inoculum size (A), experimental addition amount (B), fermentation temperature (C), and fermentation time (D)—this experiment selected four factors at three levels to further explore the optimal fermentation conditions under the influence of each factor, using L9(3) 4 An orthogonal array was used to optimize the fermentation process of pickled chili peppers using sensory evaluation indicators. The level table is shown in Table 5, and the orthogonal results are shown in Table 6.

[0061] Table 5 Orthogonal Level Table

[0062] Table 6. Results of Orthogonal Experiments

[0063] Table 7. Analysis of Variance of Orthogonal Experiments

[0064] As shown in Tables 6 and 7, based on sensory evaluation, the order of influence of the four factors on fermentation quality is D > B > C > A. Fermentation time has the greatest impact, followed by salt addition and fermentation temperature, while inoculum size has the least impact. The optimal fermentation combination is A2B2C3D3, which represents a 0.75% inoculum size, 6% salt addition, a fermentation temperature of 30℃, and a fermentation period of 30 days as the optimal fermentation process.

[0065] Using pH as an indicator, the order of influence of the four factors on fermentation quality is A > D > C > B. The inoculum size has the greatest impact, followed by fermentation time and temperature, while the amount of salt added has the least impact. The optimal fermentation combination is A3B3C3D3, which represents a 1% inoculum size, 8% salt addition, a fermentation temperature of 30℃, and a fermentation time of 30 days.

[0066] Because the combination A3B3C3D3 had excessive salt content and too large an inoculum, leading to excessively rapid fermentation and affecting the flavor and taste of the pickled chili peppers, the optimal fermentation process, A2B2C3D3, was selected after comprehensive consideration. This involved an inoculum of 0.75%, a salt content of 6%, a fermentation temperature of 30℃, and a fermentation period of 30 days. A verification experiment was conducted, and the results are shown in Table 8.

[0067] Table 8 Verification Results

[0068] In the verification experiment, the pH of A2B2C3D3 was 3.47, and the sensory score was 91. Under these conditions, the color was a uniform bright red, bright and glossy, with a fragrant fermentation aroma, harmonious smell, rich flavor, pure spiciness, mild taste, and a suitable level of saltiness, resulting in a delicious flavor. Therefore, A2B2C3D3 was selected as the optimal fermentation process.

[0069] 4. Organic Acid Detection of Chili Peppers 4.1 Methods for the detection of organic acids Using Agilent C 18 Organic acids (acetic acid, lactic acid, malic acid, tartaric acid, citric acid, succinic acid, oxalic acid, maleic acid, and fumaric acid) were detected using an AQ (4.6 mm × 250 mm, 5 μm) column high-performance liquid chromatography (HPLC) combined with external standard method. 10 g of a well-mixed pickled chili pepper sample was weighed, 10 mL of ultrapure water was added, vortexed for 2 min, ultrasonically extracted for 30 min, centrifuged at 8000 r / min for 10 min, and the supernatant was collected. The supernatant was filtered through a 0.22 μm microporous membrane before analysis. Mobile phase: A: 0.1% phosphoric acid water; B: methanol; flow rate: 0.5 mL / min; column temperature: 35℃; injection volume: 10 μL; detector: UV detector, wavelength: 210 nm; mobile phase gradient is shown in Table 9. Table 9. Detection of mobile phase gradient for organic acids

[0070] The organic acid content is calculated using the following formula: In the formula: W—The content of the target substance in the sample, in mg / kg; C—Concentration of the target analyte in the sample solution, in mg / L; C0—Concentration of the target analyte in the blank control, in mg / L; V – Constant volume, in mL; N – Dilution factor; m — the amount of sample taken, in grams.

[0071] 4.2 Instruments and Equipment 1260 High Performance Liquid Chromatograph: Agilent Technologies, Inc., USA; TGL-16M Refrigerated Centrifuge: Hunan Xiangyi Laboratory Instrument Development Co., Ltd.; Sunny Hengping FA-1004 Electronic Balance: Shanghai Sunny Hengping Scientific Instrument Co., Ltd.; F-030 Ultrasonic Extractor: Shenzhen Fuyang Technology Group Co., Ltd.

[0072] 4.3 Reagents Dipotassium hydrogen phosphate: analytical grade, Sinopharm Chemical Reagent Co., Ltd.; Acetic acid, lactic acid, malic acid, tartaric acid, citric acid, succinic acid, oxalic acid, maleic acid, fumaric acid standard: Tanmo Quality Inspection.

[0073] 4.4 Sample Pretreatment Using naturally fermented chili peppers for 30 days as the control group, and chili peppers fermented for 30 days with 0.75% fused Weissella fusarium as the experimental group (fermented chili pepper product of Example 1), the organic acid content of the chili peppers was analyzed.

[0074] 4.5 Results and Analysis Table 10 Organic acid content of pickled chili peppers under different treatments

[0075] Note: x represents concentration, y represents peak area, R 2 The coefficient of determination.

[0076] Organic acids, as important metabolites in the fermentation process of pickled chili peppers, have a significant impact on the flavor and quality of the peppers, especially their sourness. Organic acids not only inhibit the growth of harmful microorganisms but also participate in esterification reactions to form flavor esters. As shown in Table 10, five organic acids were detected in the control group and six in the experimental group. Oxalic acid, maleic acid, and succinic acid were not detected. Tartaric acid has a slightly astringent taste and a relatively strong sourness; acetic acid is extremely acidic and has a pungent sourness; fumaric acid has a strong sourness, 1.5 times that of citric acid, and also has an astringent taste. Table 10 shows that the tartaric acid content in the control group was 31.07 mg / kg, while the experimental group had a lower content of 26.21 mg / kg; the acetic acid content in the control group was 2572.31 mg / kg, while the experimental group had 2088.14 mg / kg; the fumaric acid content in the control group was also higher. This indicates that inoculation with *Westernella fusionis* fermentation can make the sourness of pickled chili peppers milder. Lactic acid and citric acid are characteristic acidic substances in pickled chili peppers. Lactic acid has a mild and strong acidity, while citric acid has a rounded and refreshing acidity. Both lactic and citric acids contribute significantly to the acidity of pickled chili peppers, resulting in a palatable sour taste. After 30 days of fermentation, the lactic acid content in the experimental group was 1.62 times that of the control group, reaching 10677.27 mg / kg; the citric acid content in the experimental group was 1191.52 mg / kg, while the control group's content was less than 3.12 mg / kg. This indicates that inoculation with *Westernella fusionis* can accelerate the production of lactic and citric acid in pickled chili peppers, thereby shortening the fermentation time. Malic acid has 1.2 times the acidity of citric acid, possessing an apple aroma, a refreshing taste, and a long aftertaste. As shown in Table 10, the experimental group had a higher malic acid content, 1.88 times that of the control group. In conclusion, inoculation with *Westernella fusionis* not only accelerates the acid production rate of pickled chili peppers but also makes the acidity of the pickled chili peppers milder and more harmonious, contributing to the improvement of the acidity quality of pickled chili peppers.

[0077] 5. Determination of volatile flavor compounds 5.1 Instruments and Equipment 7890B-7000D Gas Chromatograph and Mass Spectrometer: Agilent Technologies, Inc., USA; Sunny Hengping FA-1004 Electronic Balance: Shanghai Sunny Hengping Scientific Instruments Co., Ltd.

[0078] 5.2 Reagents Sodium chloride: analytical grade, Sinopharm Chemical Reagent Co., Ltd.; 2-Octanol: 99% purity, Shanghai Yuanye Biotechnology Co., Ltd.

[0079] 5.3 Sample Pretreatment Using naturally fermented chili peppers for 30 days as the control group, and chili peppers fermented for 30 days with 0.75% fused Weissella fusarium as the experimental group (fermented chili pepper product of Example 1), the volatile flavor compounds of the chili peppers were analyzed.

[0080] 5.4 Determination of volatile flavor compounds Raw material pretreatment: Weigh an appropriate amount of sample into a 20mL headspace vial, add internal standard 2-octanol to make its concentration for use in the instrument 5mg / L, add saturated sodium chloride, heat at 80 degrees for 30 minutes, after 30 minutes, insert the headspace microextraction needle into the headspace vial and continue heating for 30 minutes, then desorb at 250 degrees for 5 minutes.

[0081] Chromatographic conditions: HP-5MS column (30m×0.25mm×0.25μm); column temperature: initial temperature 50℃, hold for 2 min, increase to 180℃ at 5℃ / min, hold for 5 min, then increase to 250℃ at 10℃ / min, hold for 5 min; injection port temperature: 250℃; transfer line temperature: 280℃; carrier gas flow rate: 1.0 mL / min; split ratio: splitless.

[0082] Mass spectrometry conditions: Ion source temperature: 230℃, quadrupole temperature: 150℃, mass spectrometer: EI source, mode: full scan 40-600 amu.

[0083] 5.5 Qualitative and quantitative analysis of volatile flavor compounds The experimental results were compared with the NIST database, and compounds with a matching degree greater than 80 were retained. Combined with literature data, the volatile flavor substances in the samples were checked and confirmed, and the relative content of each compound was calculated using the internal standard method.

[0084] 5.6 Results and Analysis Table 11 Volatile flavor compounds and their relative contents in two types of pickled chili peppers

[0085]

[0086]

[0087] Note: "---" indicates not detected.

[0088] As shown in Table 11, a total of 84 flavor compounds were detected in the two types of pickled chili peppers, with a significant difference between the two groups. The control group detected 57 compounds, including 12 alcohols, 8 alkenes, 5 alkanes, 12 esters, 4 acids, 3 ketones, 4 aldehydes, and 9 other compounds. The experimental group detected 71 compounds, including 12 alcohols, 9 alkenes, 6 alkanes, 18 esters, 5 acids, 5 ketones, 5 aldehydes, and 11 other compounds. The flavor of pickled chili peppers is mainly composed of alcohols, esters, alkenes, and aldehydes. These flavor compounds exhibit characteristic aromas, harmonizing with each other to create the unique sour, spicy, sweet, umami, and salty flavors of pickled chili peppers. Overall, the experimental group had a richer variety of flavor compounds and a relatively higher content.

[0089] 5.6.1 Alcohols Alcohols are an important component of the aroma of pickled chili peppers, mainly synthesized through various metabolic pathways, such as amino acid metabolism and lactose metabolism. Alcohols can also react with acids to form aromatic esters. As shown in Table 11, both types of pickled chili peppers contain eucalyptol, linalool, phenethyl alcohol, 4-terpene alcohol, alpha-terpineol, citronellol, geraniol, and trans-nerolidol. Among them, eucalyptol has a eucalyptus aroma, linalool has a lily-of-the-valley or lily-like aroma, phenethyl alcohol and citronellol have a sweet rose aroma, 4-terpene alcohol has a faint earthy aroma, alpha-terpineol has a lilac aroma, geraniol has a strong sweet rose aroma, and trans-nerolidol has a citrus, floral, sweet and fresh aroma. It is worth noting that the relative content of characteristic aroma alcohols in fermented chili peppers inoculated with *Westernella fusion* was significantly higher than that in naturally fermented chili peppers. For example, the relative content of linalool in the control group was 6.481 mg / kg, while that in the experimental group was 56.439 mg / kg; the relative content of phenylethanol in the control group was 0.393 mg / kg, while that in the experimental group was 35.196 mg / kg; the relative content of citronellol in the control group was 3.289 mg / kg, while that in the experimental group was 46.525 mg / kg; and the relative content of geraniol in the control group was 29.690 mg / kg, while that in the experimental group was 121.850 mg / kg. This indicates that inoculation with *Westernella fusion* can significantly increase the relative content of aroma alcohols, giving fermented chili peppers a rich floral and honey aroma.

[0090] 5.6.2 Ester compounds Ester compounds are formed during the fermentation of pickled chili peppers through esterification reactions. Most have a fruity aroma and generally have a low olfactory threshold, contributing significantly to the flavor of pickled chili peppers and being the main flavor compounds. Table 11 shows that borneol acetate, ethyl trans-4-decenoate, (Z)-3,7-dimethyl-2,6-octadien-1-ol acetate, ethyl laurate, ethyl tetradecanoate, ethyl pentadecanoate, methyl palmitate, ethyl (Z,Z)-9,12-octadecadienoate, ethyl linoleate, and ethyl octadecanoate are ester compounds common to two types of pickled chili peppers. Among them, borneol acetate has a sweet, balsamic aroma, while trans-4-decenoate has a fruity aroma. Ethyl 4-decenoate has a waxy and pear-like aroma; (Z)-3,7-dimethyl-2,6-octadien-1-ol acetate has a citrus aroma; ethyl laurate has a mild fruity and floral aroma with a slightly oily scent; ethyl tetradecanoate has a sweet beeswax-like flavor; ethyl pentadecanoate has a honey-sweet taste; methyl palmitate has an oily, waxy iris-like aroma; and ethyl (Z,Z)-9,12-octadecadienoate has a distinctive fruity aroma. The fermented chili peppers inoculated with *Westernia fusionis* contained a higher relative content of (Z)-3,7-dimethyl-2,6-octadien-1-ol acetate, which has a citrus aroma. The relative content in the control group was 27.259 mg / kg, while that in the experimental group was 138.100 mg / kg, which was 5.07 times higher than that in the control group. The experimental group also contained 2-phenylethyl propionate, which has honey and strawberry flavors, and ethyl nonanoate, a wine-like ester compound with fruity and rose-like aromas. These components enriched the aroma of the fermented chili peppers, resulting in a better flavor and quality.

[0091] 5.6.3 Alkenes and Alkane Compounds The alkene compounds in pickled chili peppers mainly originate from the cleavage of fatty acid alkoxy radicals. They have a high flavor threshold and contribute little to the overall flavor of the pickled chili peppers. α-curcumin, shared by both types of pickled chili peppers, has a balsamic aroma; the relative content in the control group was 10.680 mg / kg, while in the experimental group it was 190.736 mg / kg. β-sesquiphellandrene, also shared, has a fruity aroma; the relative content in the control group was 11.300 mg / kg, while in the experimental group it was 236.464 mg / kg; the relative content was higher in the experimental group. Most of the detected alkane compounds are colorless and odorless, and their flavor threshold is also high, contributing little to the flavor of the pickled chili peppers.

[0092] 5.6.4 Acids, ketones, and aldehydes Acids, ketones, and aldehydes were detected in small amounts in both types of pickled chili peppers, but due to their different flavor thresholds, they play an important role in the flavor of the pickled chili peppers. Acids are mainly produced by the metabolism of lactic acid bacteria. Pickled chili peppers inoculated with *Westernella fusionis* and fermented for the same time had a greater variety and relatively higher content of acids, giving the pickled chili peppers a more tangy and refreshing characteristic. Ketones are stable and have a long-lasting aroma, generally with a pleasant vanilla or fruity flavor, but their flavor threshold is relatively high, so they do not contribute much to the flavor. Aldehydes have a stronger aroma and a lower flavor threshold, with strong lemon or floral aromas. For example, citral, which has a strong lemon aroma, had a relative content of 13.008 mg / kg in the control group and 82.103 mg / kg in the experimental group, with the experimental group having a higher relative content.

[0093] 5.6.5 Other Categories Besides alcohols and esters, pickled chili peppers also contain sulfur compounds, benzene, phenols, and other substances that significantly affect their flavor. These include diallyl disulfide, which has a strong garlic flavor; D-camphor, synthetic dextrorotatory and levorotatory borneol, and p-phenylenediamine, which has a clove-like aroma. Diallyl trisulfide, also known as allicin, is a bioactive substance derived primarily from alliin in garlic through enzymatic catalysis and thermal decomposition, possessing antibacterial and anti-inflammatory properties.

Claims

1. A fusion strain of Weissella ( Weissella confusa) R2210, characterized in that: This bacterium was deposited at the China Center for Type Culture Collection on January 29, 2024, with accession number CCTCC NO: M 2024244.

2. A fusion strain of Weissella as described in claim 1 ( Weissella confusa) Application of R2210 in fermented chili products.

3. The application according to claim 2, characterized in that: The fermentation strain used in the fermentation process of the pickled chili product includes the Fusion Weissella bacterium described in claim 1.

4. The application according to claim 2, characterized in that: The fermentation strain used in the fermentation process of the pickled chili product is the Fusion Weissella bacterium described in claim 1.

5. The application according to claim 2, characterized in that: The fused Weissella bacteria can increase the content of flavor substances in fermented chili products and accelerate the acid production rate of fermented chili.

6. The application according to claim 2, characterized in that: The preparation method of the fermented chili product is carried out according to the following steps: (1) Raw material preparation: Wash the fresh red chili peppers, remove the stems from the chili peppers, cut them into small pieces of 2-8mm, add 4-8% salt, 10-14% ginger, 10-14% garlic, 2-4% soybeans, and 1.5-3% white wine, mix well to obtain product A, and set aside. (2) Cleaning and disinfection of fermentation tank: Clean the fermentation tank, dry it at 30-60℃, sterilize and disinfect it with ultraviolet light, and set it aside for later use; (3) Preparation of protective agent: 4.0-6.0g of trehalose, 7.0-9.0g of skim milk powder and 3.0-5.0g of glycerin are added to 100mL of distilled water and mixed well to obtain the protective agent for later use; (4) Preparation of direct-inoculation starter culture: Take fusion Weissella ( Weissella confusa R2210) was added to a protectant and mixed thoroughly, then combined with Weissella foenum-graecum ( Weissella confusa The ratio of R2210 to protectant was 1g:4-6ml. The mixture was dispensed into plates and freeze-dried under vacuum to obtain a direct-inoculation fermentation agent. (5) Fermentation: Add 0.6-1.0% of direct-inoculation fermentation agent to product A, mix well, put it into a cleaned and disinfected fermentation tank, and ferment at 20-40℃ for 20-35 days. The fermentation environment is an oil-free environment, and the fermented chili product is obtained.

7. The application according to claim 6, characterized in that: The preparation method of the fermented chili product is carried out according to the following steps: (1) Raw material preparation: Wash the fresh red chili peppers, remove the stems from the chili peppers, cut them into small pieces of 3-7mm, add 5-7% salt, 11-13% ginger, 11-13% garlic, 2-3% soybeans, and 1.5-2.5% white wine, mix well to obtain product A, and set aside. (2) Cleaning and disinfection of fermentation tank: Clean the fermentation tank, dry it at 30-50℃, and sterilize and disinfect it by ultraviolet irradiation for 30-60 minutes. (3) Preparation of protective agent: Take 4.5-5.5g of trehalose, 7.5-8.5g of skim milk powder and 3.5-4.5g of glycerin, add them to 100mL of distilled water, mix well and the protective agent is obtained for later use; (4) Preparation of direct-inoculation starter culture: Take fusion Weissella ( Weissella confusa R2210) was added to a protectant and mixed thoroughly, then combined with Weissella foenum-graecum ( Weissella confusa The ratio of R2210 to the protectant was 1g:4.5-5.5ml. The mixture was dispensed into plates and then freeze-dried under vacuum to obtain a direct-inoculation fermentation agent. (5) Fermentation: Add 0.7-0.85% of direct-inoculation fermentation agent to product A, mix well, put it into a cleaned and disinfected fermentation tank, and ferment at 25-35℃ for 25-30 days. The fermentation environment is an oil-free environment, and the fermented chili product is obtained.

8. The application according to claim 7, characterized in that: The preparation method of the fermented chili product is carried out according to the following steps: (1) Raw material preparation: Wash the fresh red chili peppers, remove the stems from the chili peppers, cut them into small pieces of 4-6mm, add 6% salt, 12% ginger, 12% garlic, 2.5% soybeans and 2% white wine, mix well to obtain product A, and set aside. (2) Cleaning and disinfection of fermentation tank: Clean the fermentation tank, dry it at 40℃, and sterilize and disinfect it by ultraviolet irradiation for 30-60 minutes. (3) Preparation of protective agent: Take 5.0g of trehalose, 8.0g of skim milk powder and 4.0g of glycerin, add them to 100mL of distilled water, mix well to obtain protective agent, and set aside; (4) Preparation of direct-inoculation starter culture: Take fusion Weissella ( Weissella confusa R2210) was added to a protectant and mixed thoroughly, then combined with Weissella foenum-graecum ( Weissella confusa R2210) was mixed with a protectant at a ratio of 1g:5ml, dispensed into plates, and then freeze-dried under vacuum to obtain a direct-inoculation fermentation agent. (5) Fermentation: Add 0.75% direct-inoculation fermentation agent to product A, mix well, put it into a cleaned and disinfected fermentation tank, ferment at 30℃ for 30 days, and the fermentation environment is an oil-free environment to obtain the fermented chili product.

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