Lactobacillus fermentum and application thereof in degrading zanthoxyli perfractum

Abstract

This invention provides a strain of *Lactobacillus fermentatus* and its applications, belonging to the field of microbial technology. The *Lactobacillus fermentatus* strain is named *Lactobacillus fermentatus* JMS-ZY and is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 37637. This invention also provides the application of this strain in reducing xanthophoretic compounds in Sichuan pepper seed meal. Specifically, activated *Lactobacillus fermentatus* JMS-ZY seed culture is inoculated into raw Sichuan pepper seed meal, and the moisture content and inoculation amount are adjusted. Solid-state fermentation is carried out at a suitable temperature. After fermentation, the product is dried and pulverized to obtain Sichuan pepper seed meal with low xanthophoretic flavor. This invention utilizes a specific *Lactobacillus fermentatus* strain for solid-state fermentation of Sichuan pepper seed meal, which can effectively degrade xanthophoretic compounds, thereby improving the palatability of the Sichuan pepper seed meal and increasing its utilization rate as a feed ingredient. This method is simple, green, and safe, providing a new approach for the resource utilization of Sichuan pepper seed meal.

Description

Technical Field

[0001] This invention belongs to the field of microbial fermentation technology, specifically relating to a strain of fermenting lactobacillus and its application in the degradation of zanthoxylum bungeanum. Background Technology

[0002] Sichuan pepper contains abundant numbing substances, among which amide compounds are the characteristic components of its numbing flavor, and their content is one of the main indicators determining the quality of Sichuan pepper. These numbing substances are widely distributed in the stems, leaves, pericarp, and seeds, with the pericarp containing the highest concentration (Zhang et al., 2022). In recent years, researchers have increasingly confirmed that the key irritant element in Sichuan pepper is alkylamides, represented by sanshool, which are mostly chain-like unsaturated fatty amides and amides linked to aromatic rings. Currently, more than 20 amide substances have been reported in Sichuan pepper, mainly α-sanshool, β-sanshool, γ-sanshool, hydroxy-α-sanshool, hydroxy-β-sanshool, and hydroxy-γ-sanshool. Among them, hydroxy-α-sanshool and hydroxy-β-sanshool are considered to be the main numbing substances, and their content can be regarded as an important indicator for evaluating the quality of Sichuan pepper (Ma et al., 2025; Shi et al., 2025; Wang et al., 2024).

[0003] Because the numbing and aromatic compounds in Sichuan pepper are unstable, structurally similar, and diverse, they are difficult to detect. Currently, the main methods for detecting these compounds include high-performance liquid chromatography (HPLC), thin-layer chromatography (TLC), ultraviolet spectrophotometry (UV), gas chromatography-mass spectrometry (GC-MS), infrared spectroscopy (IR), and rapid detection methods based on formaldehyde titration (FT). Among these methods, HPLC has advantages such as stable detection results and low detection limits (Luo et al., 2022b). (Wang et al., 2024) used HPLC to determine the content of numbing and aromatic components in 16 varieties of Sichuan pepper from different origins. The results showed significant differences in the content of hydroxy-α-sanshool and hydroxy-β-sanshool among different types of Sichuan pepper. The content of hydroxy-α-sanshool ranged from 60.06±1.14 to 164.13±3.28 mg / g, and the content of hydroxy-β-sanshool ranged from 7.81±0.36 to 21.11±0.75 mg / g. The RSDs of hydroxy-α-sanshool ranged from 1.73% to 3.80%, and those of hydroxy-β-sanshool ranged from 2.03% to 4.73% (RSDs ≤ 5%), indicating that the measurements of hydroxy-α-sanshool and hydroxy-β-sanshool are reliable. (Xie Wangjun et al., 2014) Sichuan pepper is a traditional Chinese spice and medicinal plant. During its intensive processing, its byproducts contain abundant nutrients such as protein and oil, possessing high development and utilization value (Yuan Congjun et al., 2022). Sichuan pepper seed meal is a major byproduct of Sichuan pepper oil processing, rich in protein, fat, and various bioactive substances, showing high potential for feed development. However, due to its content of alkylamide-based schizocarpine, which is both highly irritating and toxic, its palatability is affected, limiting its application in animal feed and also causing environmental pollution.

[0004] To achieve the safe and high-value utilization of Sichuan pepper seed meal, a systematic study was conducted focusing on "functional bacteria screening, fermentation process optimization, nutritional value enhancement, and application effect evaluation." The study was validated by screening microorganisms that may degrade Sichuan pepper glycosides, and finally a strain of fermenting lactobacillus that can effectively degrade Sichuan pepper glycosides was obtained. Summary of the Invention

[0005] This invention screened a strain of *Lactobacillus fermentum* from the environment. The strain is named *Lactobacillus fermentum* JMS-ZY and was deposited on February 2, 2026, at the China General Microbiological Culture Collection Center (CGMCC). The accession number for this strain is CGMCC No. 37637. Latin name: Lactobacillus fermentum Address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0006] This strain possesses the physiological ability to specifically degrade zestin, and can stably colonize and function in a solid-state fermentation system with zest seed meal as the main substrate. Based on the above-mentioned strain, the present invention provides the application of this strain in reducing xanthophorin in pepper seed meal. Specifically, the fermenting lactobacillus JMS-ZY is used to carry out solid-state fermentation of raw pepper seed meal to reduce xanthophorin.

[0007] The specific fermentation steps are as follows: (1) Seed culture preparation: The preserved Lactobacillus fermentum JMS-ZY was activated by streaking on MRS agar medium, and a single colony was picked and inoculated into MRS liquid medium. The culture was carried out at 35~40℃ for 20~22 hours until the strain was in the logarithmic growth phase to obtain the seed culture.

[0008] (2) Solid-state fermentation: The seed liquid obtained in step (1) is inoculated into the raw material of Sichuan pepper seed meal, and sterile water is added at the same time to adjust the initial moisture content to 35%~45%, and the inoculation amount is controlled to be 4%~8% (v / w). After thorough mixing, it is placed under a constant temperature of 30~40℃ for solid-state fermentation for 48~96 hours. During the fermentation period, in order to ensure uniform fermentation of the material and ventilation and heat dissipation, the material is turned over once every 12 hours.

[0009] (3) Post-processing: After fermentation, the wet material is taken out and dried at a low temperature of 40~48℃. Finally, it is crushed and passed through a 20~100 mesh sieve to obtain fermented pepper seed meal product with significantly reduced pepper sesame content. Beneficial effects

[0010] First, this invention screened a strain of *Lactobacillus fermentum* JMS-ZY that can effectively degrade trichosanthes in various environments. From the perspective of strain growth characteristics, trichosanthes generally has a certain inhibitory effect on the growth of various strains. However, the *Lactobacillus fermentum* JMS-ZY strain preserved in this invention can still maintain stable growth under the presence of trichosanthes, indicating that it has good tolerance to trichosanthes. Its mechanism for degrading trichosanthes may include amide structure transformation, conjugated double bond breakage, and chemical degradation caused by pH changes during fermentation. Further research is needed to determine the specific mechanism.

[0011] Secondly, the fermenting lactobacillus JMS-ZY provided by this invention has a good degradation effect on zanthoxylin, especially hydroxy-α-santhiolin and hydroxy-β-santhiolin. After applying this strain to the fermentation treatment of Sichuan pepper seed meal, the zanthoxylin content in Sichuan pepper seed meal can be reduced to the range that meets the requirements for feed addition in a short time. Moreover, the treatment process is simple and the cost is low, making it suitable for large-scale promotion and application.

[0012] Furthermore, *Lactobacillus fermentum* JMS-ZY exhibits high degradation efficiency of trichosanthesin under mild fermentation conditions. The optimal fermentation conditions were screened using single-factor and orthogonal experiments. Under these optimal conditions, *Lactobacillus fermentum* JMS-ZY significantly reduces the trichosanthesin content in Sichuan pepper seed meal, effectively removing its irritant properties. This addresses the limitations of Sichuan pepper seed meal in feed applications, improves its utilization rate as a feed ingredient, and achieves high-value utilization of Sichuan pepper by-products while reducing environmental pollution from industry processing waste. This approach combines economic and environmental benefits, providing a new microbial resource and technological pathway for the high-value utilization of Sichuan pepper by-products. Validation experiments under the optimal combination conditions showed a trichosanthesin degradation rate of 42.21%, slightly higher than the 41.20% in the orthogonal experiment, indicating good repeatability and stability of the optimization results. Similar phenomena are common in fermentation process optimization studies, usually related to more stable fermentation environment control and reduced operational errors in validation experiments. The validation results are consistent with the predicted values, further demonstrating the reliability of the determined process parameters.

[0013] Finally, this study screened and obtained a fermenting lactobacillus strain with strong ability to degrade zestin, providing a strain basis for the biological detoxification and feed development of zest seed meal. Future research will focus on the application effect of this strain in zest seed meal fermentation and optimize fermentation conditions. Attached Figure Description

[0014] Figure 1 Colony and strain morphology of Lactobacillus fermentum JMS-ZY on MRS medium.

[0015] Figure 2 Figure: Homology analysis results of Lactobacillus fermentum JMS-ZY Blast.

[0016] Figure 3 Growth curve of Lactobacillus fermentum JMS-ZY.

[0017] Figure 4 Factor level table for single-factor experimental design.

[0018] Figure 5 Effect of fermentation temperature on the degradation rate of zanthoxylum bungeanum in zanthoxylum bungeanum seed meal.

[0019] Figure 6 Effect of fermentation time on the degradation rate of zanthoxylum bungeanum in zanthoxylum bungeanum seed meal.

[0020] Figure 7 Effect of inoculation amount on the degradation rate of zanthoxylum bungeanum in zanthoxylum bungeanum seed meal.

[0021] Figure 8 Effect of moisture content on the degradation rate of zanthoxylum bungeanum in zanthoxylum bungeanum seed meal.

[0022] Figure 9 Table of results and range analysis of orthogonal experiments on single-strain solid-state fermentation.

[0023] Figure 10 Results of optimal fermentation process for Sichuan pepper seed meal. Detailed Implementation

[0024] The present invention will now be described in detail with reference to the embodiments and accompanying drawings.

[0025] In this embodiment, the sesame extract was purchased from Hubei Langbowan Biomedical Co., Ltd. The pepper seed meal, a byproduct of pepper oil processing, was sourced from the Hebei Zunhua Experimental Base of the Chinese Academy of Agricultural Sciences. Before use, it was crushed and passed through a 40-mesh sieve, then sealed and stored for later use.

[0026] MRS medium (g / L): peptone 10, yeast extract 5, beef extract 10, glucose 20, dipotassium hydrogen phosphate 2, sodium acetate 5, triammonium citrate 2, magnesium sulfate 0.1, manganese sulfate monohydrate 0.05, Tween 80 1, sterilized at 121℃ for 20 min.

[0027] MRS agar medium (g / L): peptone 10, yeast extract 5, beef extract 10, glucose 20, dipotassium hydrogen phosphate 2, sodium acetate 5, triammonium citrate 2, magnesium sulfate 0.1, manganese sulfate monohydrate 0.05, Tween 80 1, agar 15, sterilized at 121℃ for 20 min.

[0028] MRS calcium carbonate agar medium (g / L): peptone 10, yeast extract 5, beef extract 10, glucose 20, dipotassium hydrogen phosphate 2, sodium acetate 5, triammonium citrate 2, magnesium sulfate 0.1, manganese sulfate monohydrate 0.05, Tween 80 1, calcium carbonate 10, sterilized at 121℃ for 20 min.

[0029] In this embodiment, the method for determining the content of zanthoxylin was high-performance liquid chromatography, and the specific operation is as follows: Preparation of standard solutions: Take 10 mg of hydroxy-α-sanshool reference standard (accurate to 0.1 mg), place it in a beaker, dissolve it in chromatographic grade methanol, and make up to 10 mL; dilute with chromatographic grade methanol to prepare working solutions of reference standard with concentrations of 5 μg / mL, 10 μg / mL, 20 μg / mL, 30 μg / mL, 40 μg / mL, and 50 μg / mL, for testing.

[0030] Chromatographic conditions: An Agilent XDB-C18 column (250 mm × 4.6 mm, 5 µm) was used. The mobile phase consisted of water and acetonitrile (55:45, V / V), with a gradient elution program. The detection wavelength was 270 nm, the column temperature was set at 40°C, the flow rate was 0.8 mL / min, and the injection volume was 10 µL.

[0031] Sample preparation: Weigh 1g (accurate to 0.001g) of Sichuan pepper seed meal into a 50mL centrifuge tube, add 20mL of analytical grade methanol and seal the tube. Extract the numbing and spicy substances by ultrasonication (45℃, 360W, 40kHz, 40min), centrifuge (2500r / min, 10min), transfer the supernatant to a 100mL volumetric flask, repeat the above operation twice, make up to volume with methanol, store at -20℃ for later use, filter through a 0.22μm organic microporous membrane, and use for liquid chromatography analysis. Example 1

[0032] A strain of *Lactobacillus fermentum* JMS-ZY, which was deposited on February 2, 2026, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 37637, has the Latin name: Lactobacillus fermentum Figure 2 Address: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0033] The method for isolating the Lactobacillus fermentum JMS-ZY is as follows: ① Using soil from citrus orchards and other orchards, humus layers with minimal human activity, rotten wood, macrofungi, various fermented foods, rotten haystacks, rumen fluid, and animal feces as bacterial sources, strain enrichment solutions were prepared for each sample using the dilution-mixing method. Accurately weigh 10 g or 10 mL of each sample and add it to an Erlenmeyer flask containing 90 mL of sterile physiological saline with glass beads. Shake at 200 r / min for 20 min to thoroughly mix the sample and disperse the bacteria. Use a pipette to transfer the supernatant suspension to a sterile centrifuge tube for later use.

[0034] ② The above suspension was serially diluted with sterile physiological saline. 1 ml of each solution was inoculated into MRS medium for enrichment and culture for 24 hours. The bacterial suspension was then spread onto MRS calcium carbonate agar and incubated at 37°C for 48 hours. Single colonies producing clear zones, but differing in size, morphology, and color, were selected for isolation and purification. All treatments were performed in triplicate. The purified strains were stored in 50% glycerol at -20°C.

[0035] ③ After activating the colonies obtained in step ②, they were inoculated into the primary screening medium for trichosanthesin, incubated at 37℃ for 48 h, centrifuged at 3000 rpm for 15 min, and the supernatant was taken to determine the trichosanthesin content. Strains that can degrade trichosanthesin were identified as candidate strains.

[0036] The initial screening medium for Sichuan peppercorn extract (g / L) consisted of: peptone 10, yeast extract 5, beef extract 10, glucose 20, dipotassium hydrogen phosphate 2, sodium acetate 5, triammonium citrate 2, magnesium sulfate 0.1, manganese sulfate monohydrate 0.05, Tween 80, with an appropriate amount of Sichuan peppercorn extract added, and sterilized at 121℃ for 20 min.

[0037] ④ The strains obtained from the initial screening were activated and inoculated into the secondary screening medium at an inoculum rate of 2%. The cultures were incubated at 37℃ for 48 h, centrifuged at 3000 r / min for 15 min, and the supernatant was collected to determine the content of xanthocyanin. Three replicates were performed for each strain. Strains capable of degrading xanthocyanin were obtained.

[0038] Molecular biological identification of the strain's 16S rDNA: DNA from the target strain was extracted using a bacterial genomic DNA extraction kit. Universal primers for 16S rDNA were used. (27F: 5'-AGAGTTTGATCCTGGCTCAG-3'; 1492R: 5'-CTACGGCTACCTTGTTACGA-3') 16S rDNA of the strain was amplified. After amplification, the purified PCR product was sent to Qingdao Qingke Biotechnology Co., Ltd. for sequencing, and the obtained sequence was submitted to the NCBI GenBank database. Homology comparison analysis was performed using the BLAST program, and the results are as follows: Limosilactobacillus fermentum As shown. The results showed that the 16S rRNA gene sequence of strain R1B5-1 was highly similar to several type strains of Limosilactobacillus fermentum, among which... Limosilactobacillus fermentum The sequence homology of strain CIP 102980 reached 100.00%, and was consistent with... Limosilactobacillus fermentum The similarity with strain NBRC 15885 was 99.91%, with an E-value of 0.0 and a coverage of 98%. Based on the results of 16S rRNA gene sequence homology analysis, combined with the colony morphology and Gram staining characteristics of the strain, strain JMS-ZY was preliminarily identified as *Lactobacillus fermentum*. Figure 3 ).

[0039] Observation of bacterial morphology: The target bacteria were inoculated onto MRS solid medium using the plate division method and incubated at 37°C for 24 hours. The morphology, color, edge uniformity, and surface texture of the bacteria were observed under a microscope. Subsequently, the bacteria were cultured in MRS liquid medium at 37°C for 24 hours, and the morphology of the target bacteria was observed under a microscope using Gram staining.

[0040] The strain JMS-ZY was inoculated onto MRS solid medium and incubated at 37 ℃ for 24 h. Colony morphology was observed under a magnifying glass, as shown in Figure 1: the colonies were milky white, approximately 1-2 mm in diameter, with a smooth, moist surface, regular and opaque edges, and were generally round or nearly round with no obvious central protrusion, consistent with the morphological characteristics of Lactobacillus colonies. After culturing the strain in MRS liquid medium with shaking for 24 h, Gram staining was performed according to the instructions. The results are shown in Figure 1: after staining, the strain appeared as purple, slender or short rods. Therefore, strain JMS-ZY was preliminarily identified as a Gram-positive Lactobacillus, with morphological characteristics consistent with Lactobacillus fermentum.

[0041] Determination of the growth curve of *Lactobacillus fermentum* JMS-ZY: The isolated target strain was streaked onto plates. Single colonies from the plates were picked and incubated in 10 ml of liquid MRS medium at 37°C for 24 h to prepare a seed culture. This seed culture was then inoculated at a rate of 1% into 150 ml of liquid MRS medium and incubated statically at 37°C for 48 h. Samples were taken every 3 h, and the optical density (OD) was measured at 600 nm using a UV-Vis spectrophotometer. An OD600 versus time curve was plotted. Results are as follows: Figure 3 As shown.

[0042] Depend on Figure 4 It can be seen that the growth curve of Lactobacillus fermentum is in the lag phase from 0 to 2 h, with low growth of the strain; it is in the logarithmic growth phase from 2 to 8 h, when the growth rate is the fastest; the growth slows down from 8 to 12 h, entering the stationary phase; the growth of the strain remains basically stable from 12 to 24 h, with the OD value of the strain reaching its maximum from 20 to 22 h. Therefore, in subsequent culture and fermentation experiments, activated bacterial solution cultured for 20 to 22 h was selected as the seed culture. Example 2

[0043] The strain in this embodiment is Lactobacillus fermentum JMS-ZY, which has the ability to degrade Zanthoxylum bungeanum, screened and preserved in Example 1. Before use, it was activated in MRS liquid medium at 37 °C for 20-22 h to bring it into the logarithmic growth phase as the fermentation seed liquid.

[0044] The method for preparing the fermentation seed culture is as follows: the preserved Lactobacillus fermentum JMS-ZY was streaked on MRS agar medium and cultured at 37℃ for 24 h. A single colony was picked from the plate and placed in 50 mL of MRS liquid medium and cultured at 37℃ for 22 h to prepare the Lactobacillus fermentum JMS-ZY seed culture.

[0045] Weigh 50 g of Sichuan pepper seed meal into a 250 mL beaker, and add the fermented Lactobacillus JMS-ZY seed culture according to... Figure 4 The initial moisture content was adjusted by adding sterile water to the inoculation ratio. After thorough mixing, the mixture was added to the fermentation substrate (Sichuan pepper seed meal raw material) according to the inoculation amount. After inoculation, the mixture was thoroughly stirred with a sterile glass rod to ensure uniform mixing of the bacterial solution and the substrate, and then sealed with a breathable membrane. The treated sample was placed under constant temperature for solid-state fermentation for 72 hours. During fermentation, the sample was turned over every 6-12 hours to ensure the homogeneity of the fermentation system. After fermentation, the sample was dried at 40-48℃, then pulverized and passed through a 20-100 mesh sieve, sealed and stored for the determination of Sichuan pepper sesame content.

[0046] The content of zanthoxylin was determined by high performance liquid chromatography (HPLC). Using the zanthoxylin content in unfermented zanthoxylin seed meal as the initial value, the degradation rate of zanthoxylin under different treatment conditions was calculated.

[0047] The degradation rate of Sichuan pepperin is calculated using the following formula: Degradation rate (%) = (Initial xanthocyanin content - Xanthocyanin content after fermentation) / Initial xanthocyanin content × 100% The *Lactobacillus fermentum* JMS-ZY culture broth was prepared into an inoculum of the required concentration and inoculated into the *Zanthoxylum bungeanum* seed meal fermentation substrate according to the set ratio. The substrate was sealed with a respiratory membrane and allowed to ferment statically at a constant temperature. Under the same basic conditions, the effects of fermentation time, fermentation temperature, inoculum size, and initial moisture content on the degradation of trichosanthesin were investigated. The fermentation time was set at 2, 3, and 4 days; the fermentation temperature at 30 ℃, 37 ℃, and 40 ℃; the inoculum size at 4%, 6%, and 8%; and the initial moisture content at 35%, 40%, and 45%. The degradation rate of trichosanthesin was used as the evaluation index. Each treatment was repeated in triplicate. Specific experimental groupings are detailed below. Figure 6 .

[0048] Figure 5 shows the effect of *Lactobacillus fermentatus* on the degradation rate of trichosanthes in *Zanthoxylum bungeanum* seed meal under different fermentation temperatures. As shown in the figure, the degradation rate of trichosanthes first increased and then decreased with increasing fermentation temperature. At a fermentation temperature of 37 ℃, the degradation rate of trichosanthes was significantly higher than that of the 30 ℃ and 40 ℃ treatment groups (P<0.05), reaching 35.41%. The difference between the 30 ℃ and 40 ℃ treatment groups was significant (P<0.05), with the 40 ℃ treatment group showing a higher degradation rate than the 30 ℃ treatment group. In summary, fermentation at 37 ℃ yielded the best results under all treatment conditions; therefore, 37 ℃ was determined to be the optimal fermentation temperature.

[0049] Figure 7 This study investigated the effect of different fermentation times using *Lactobacillus fermentum* JMS-ZY on the degradation rate of xanthocyanin in *Zanthoxylum bungeanum* seed meal. The results showed that the degradation rate of xanthocyanin generally increased and then decreased with prolonged fermentation time. At a fermentation time of 72 h, the degradation rate of xanthocyanin was significantly higher than other treatment groups (P<0.05), reaching a maximum of 34.81%. While there was no significant difference between the 48 h and 96 h treatment groups (P>0.05), both were significantly lower than the 72 h treatment group (P<0.05). Therefore, 72 h was determined to be the optimal fermentation time.

[0050] Figure 8The effect of different inoculum amounts of *Lactobacillus fermentum* JMS-ZY on the degradation rate of trichosanthes in *Zanthoxylum bungeanum* seed meal was shown in the figure. As the inoculum amount increased, the degradation rate of trichosanthes first increased and then decreased. At an inoculum amount of 6%, the degradation rate of trichosanthes was 29.96%, significantly higher than that of the 4% and 8% inoculum groups (P<0.05). However, there was no significant difference between the 4% and 8% inoculum groups (P>0.05), and both were lower than the 6% inoculum group (P<0.05). Therefore, 6% was determined to be the more suitable inoculum amount.

[0051] Figure 9 This study investigated the effect of *Lactobacillus fermentum* JMS-ZY on the degradation rate of xanthophyll in *Zanthoxylum bungeanum* seed meal under different initial moisture contents. The results showed that the degradation rate of xanthophyll gradually increased with increasing initial moisture content. At an initial moisture content of 45%, the degradation rate of xanthophyll was significantly higher than that of the 35% and 40% treatment groups (P<0.05), reaching a maximum of 22.14%; and the 40% treatment group was significantly higher than the 35% treatment group (P<0.05). Therefore, under the conditions of this experiment, 45% was determined to be the suitable initial moisture content.

[0052] Based on single-factor experiments, and using the degradation rate of xanthophyll in Sichuan pepper seed meal as an evaluation index, four factors—fermentation temperature, fermentation time, inoculum size, and initial moisture content—were selected. A four-factor, three-level orthogonal experiment was conducted to optimize the single-strain solid-state fermentation process conditions. The results of the orthogonal experiment are shown below. Figure 9 .

[0053] pass Figure 10 It can be seen that the degradation rate of trichosanthesin varies under different treatment combinations, with the highest degradation rate (41.20%) observed in group 5, indicating that changes in fermentation process parameters affect the degradation effect of trichosanthesin. Range analysis shows that the K values ​​of each factor differ at different levels. Based on the range (R) values, the order of influence of each factor on the degradation rate of trichosanthesin is: fermentation temperature > inoculum size > initial moisture content > fermentation time. Among these, fermentation temperature has the most significant impact on the degradation rate of trichosanthesin, followed by inoculum size and initial moisture content, while the effect of fermentation time is relatively weak. Further comparison of the K values ​​at different levels of each factor shows that the degradation effect of trichosanthesin is best when the fermentation temperature is 37℃, the inoculum size is 6%, the initial moisture content is 35%, and the fermentation time is 72h. The range analysis results indicate that the optimal process for single-strain fermentation of Sichuan pepper seed meal is: fermentation temperature 37℃, inoculum size 6%, initial moisture content 35%, and fermentation time 72h.

[0054] Range analysis showed that the influence of various factors on the degradation rate of trichosanthesin varied, with fermentation temperature having the greatest impact, followed by inoculum size, initial moisture content, and fermentation time. Based on K-value and R-value analysis, the optimal fermentation combination was determined to be a fermentation temperature of 37 ℃, an inoculum size of 6%, an initial moisture content of 35%, and a fermentation time of 72 h. Under this combination of conditions, the highest degradation rate of trichosanthesin in the orthogonal experiment was 41.20%. To verify the reliability of the optimization results, a repeat validation experiment was conducted under the above optimal conditions, achieving a degradation rate of 42.21%, as shown in the figure. ​ The results showed a trend consistent with orthogonal experimental results and a slight improvement, indicating that the selected fermentation process parameters were stable, feasible, and had good repeatability.

Claims

1. A strain of Lactobacillus fermentum JMS-ZY, characterized in that, Latin name: Lactobacillus fermentum It is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC No. 37637.

2. The use of the Lactobacillus fermentum according to claim 1, characterized in that: The fermenting lactobacillus JMS-ZY is used to ferment pepper seed meal and degrade the pepperin in the pepper seed meal.

3. A method for reducing xanthocyanin in Sichuan pepper seed meal, characterized in that, The method for reducing the trichosanthes in pepper seed meal specifically involves solid-state fermentation of raw pepper seed meal using Lactobacillus fermentum JMS-ZY. The Lactobacillus fermentum JMS-ZY is deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC No. 37637.

4. The method for reducing xanthocyanin in Sichuan pepper seed meal according to claim 3, characterized in that, The specific steps of the solid-state fermentation are as follows: (1) The fermenting Lactobacillus JMS-ZY was activated to prepare a seed culture; (2) The seed liquid is inoculated into the raw material of Sichuan pepper seed meal, and sterile water is added to adjust the initial moisture content to 35-45%. The inoculation amount is 4-8%, and solid fermentation is carried out at 30-40℃ for 48-96 hours. (3) After fermentation, the material is dried and crushed to obtain fermented pepper seed meal with reduced pepper sesame content.

5. The method for reducing xanthocyanin in Sichuan pepper seed meal according to claim 4, characterized in that, The method for preparing the seed culture in step (1) is as follows: streak the preserved Lactobacillus fermentum JMS-ZY on MRS agar medium, pick a single colony and inoculate it into MRS liquid medium, and culture it at 35~40℃ for 20~22h to obtain the seed culture.

6. The method for reducing xanthocyanin in Sichuan pepper seed meal according to claim 4, characterized in that, In step (2), the material is turned over once every 12 hours during the fermentation process.

7. The method for reducing xanthocyanin in Sichuan pepper seed meal according to claim 4, characterized in that, In step (3), the drying conditions are drying at 40~48℃; the pulverization is passing through a 20~100 mesh sieve.

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