A portuguese williomia cylindracea cypj1 and application thereof

Abstract

This invention discloses a strain of *Corynebacterium lucida* CYPJ1 and its applications. The *Corynebacterium lucida* CYPJ1 has the preservation number CGMCC No. 35719, and its saccharifying enzyme activity is 73.19 ± 0.09 U / mL. Using homogenized raw materials and fermentation with this strain results in increased soluble dietary fiber content and enhanced physical and functional properties. Foods or health products fermented with homogenized raw materials and this strain have lipid-lowering effects. This invention also found that using *Corynebacterium lucida* (…) after homogenization of raw materials… Clavispora lusitaniae CYPJ1 and Kluyveromyces macrocephala ( Kluyveromyces marxianus SXJ2 or Bacillus sicca ( Bacillus siamensis The DL-RXL01 combination fermentation yields even better results.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, specifically relating to a Portuguese yeast CYPJ1 and its applications. Background Technology

[0002] Dietary fiber mainly comes from plant-based foods and is a type of carbohydrate that cannot be broken down and absorbed by enzymes in the human digestive tract. Based on its water solubility, dietary fiber can be divided into two main categories: soluble dietary fiber and insoluble dietary fiber. The health benefits of dietary fiber are extensive and profound, mainly reflected in the following aspects: (1) Promoting digestive health: Dietary fiber can absorb water and swell, stimulate the secretion of mucus in the intestines, promote intestinal peristalsis, and effectively prevent and relieve constipation. At the same time, it acts as an intestinal "cleaner," helping to dilute and accelerate the excretion of potential carcinogens and reduce the risk of colon cancer. (2) Maintaining cardiovascular health: Dietary fiber is a "guardian" of the heart. It can form a gel-like substance in the intestines, delaying the absorption of sugar and helping to stabilize blood sugar; and it can combine with bile acids to promote cholesterol excretion, thereby effectively reducing the level of total cholesterol and low-density lipoprotein in the blood and reducing the risk of atherosclerosis and coronary heart disease. (3) Regulating weight and blood sugar: Dietary fiber foods have a high water-holding capacity and low energy density, which can enhance satiety, reduce overall calorie intake, and help with weight management. Its ability to delay sugar absorption is particularly important for diabetic patients and is a powerful tool for stabilizing postprandial blood sugar. (4) Nourishing intestinal flora: Soluble dietary fiber (such as resistant starch and inulin) is a high-quality prebiotic that can serve as "nutrients" for beneficial intestinal flora, promoting their growth and reproduction, maintaining the balance of intestinal microecology, enhancing immunity, and even having a positive impact on emotional health.

[0003] Soluble dietary fiber is more beneficial to the human body than insoluble dietary fiber. Generally, the dietary fiber content in plants is higher insoluble fiber than soluble fiber. Rice bran is a mixture of the pericarp, seed coat, outer endosperm, and embryo that are removed during the refining process of brown rice into white rice. It typically accounts for 5-8% of the total weight of rice, but it concentrates more than 60% of the rice's nutrients. Rice bran contains a relatively high amount of dietary fiber, with a higher content of insoluble fiber than soluble fiber. How to convert the insoluble dietary fiber in rice bran into soluble fiber and increase the content of soluble dietary fiber is a worthy research topic in this field. Summary of the Invention

[0004] The purpose of this invention is to provide a Portuguese yeast CYPJ1 and its applications.

[0005] A type of *C. pekinensis*, classified as *C. pekinensis* (…). Clavispora lusitaniae CYPJ1, with accession number CGMCC No.35719.

[0006] The Portuguese yeast ( Clavispora lusitaniae Application of CYPJ1 in glycan-producing enzymes.

[0007] The Portuguese yeast ( Clavispora lusitaniae Application of CYPJ1 in the preparation of dietary fiber.

[0008] A compound microbial agent, including *Corynebacterium lucida* ( Clavispora lusitaniae CYPJ1 and Kluyveromyces maculae ( Kluyveromyces marxianus SXJ2; or Portuguese yeast ( Clavispora Portuguese CYPJ1 and Bacillus sicca ( Siamese Bacillus DL-RXL01.

[0009] The Max Kluyveromyces ( Kluyveromyces marxianus The SXJ2 accession number is CGMCC No. 14274; the Bacillus sicca described is... Siamese Bacillus The accession number for DL-RXL01 is CGMCC No. 23532. All of the above strains were purchased from the China General Microbiological Culture Collection Center.

[0010] The application of the compound microbial agent in the preparation of dietary fiber.

[0011] The Portuguese yeast described in this invention ( Clavispora lusitaniae CYPJ1 was deposited on August 22, 2025 at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC No. 35719.

[0012] Beneficial effects of the present invention: The present invention discovers a strain of Portuguese yeast ( Clavispora lusitaniae CYPJ1, with a saccharifying enzyme activity of 73.19±0.09 U / mL. Using homogenized raw materials and fermentation with this strain resulted in increased soluble dietary fiber content and enhanced physical and functional properties. This invention also found that homogenized raw materials, followed by the use of *Corynebacterium tumefaciens* (C. tumefaciens), resulted in improved soluble dietary fiber content and enhanced physical and functional properties. Clavispora lusitaniae CYPJ1 and Kluyveromyces maculae ( Kluyveromyces marxianus SXJ2 or Bacillus sicca ( Siamese Bacillus The DL-RXL01 combination fermentation yields even better results. Attached Figure Description

[0013] Figure 1 Images of colonies and Gram staining of *Cercospora lucida* CYPJ1.

[0014] Figure 2Phylogenetic tree of Corynebacterium lucida CYPJ1.

[0015] Figure 3 The figures show the results of the measurement of changes in mouse body weight and the measurement of weight gain; in the figure, A represents the measurement result of changes in body weight, and B represents the measurement result of weight gain.

[0016] Figure 4 The results show the measurement of lipid-related indicators in mice; in the figure, A represents TC content, B represents TG content, C represents HDL-C content, and D represents LDL-C content.

[0017] Figure 5 The results show the measurement of liver lipid-related indicators in mice; in the figure, A represents TC content, B represents TG content, C represents HDL-C content, and D represents LDL-C content.

[0018] Figure 6 The results show the measurement of liver function-related indicators in mice; in the figure, A represents AST activity and B represents ALT activity. Detailed Implementation

[0019] To facilitate understanding of the present invention, a more comprehensive description will be given below. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0020] Example 1: Strain Screening and Identification

[0021] (1) Strain screening and isolation and purification

[0022] Experimental materials: Korean kimchi from Northeast China

[0023] LB medium: glucose 3 g / L, yeast extract 3 g / L, peptone 10 g / L, sodium chloride 5 g / L. Adjust the pH to 6.3 and autoclave at 121°C for 20 minutes. (Solid medium is liquid medium with 18 g / L agar added).

[0024] Serial dilution and plating: Add 10 mL of sterile physiological saline to the kimchi juice, mix well, and then add 1 mL of the mixture to 9 mL of sterile physiological saline to prepare bacterial solutions of different concentrations. Take the three concentrations of diluted bacterial solutions, place them in solid culture medium, and spread them evenly on plates in one direction using a plating stick. Place the plate in a constant temperature incubator and incubate at 25°C for 4 days.

[0025] Streak purification: Pick individual colonies formed on the culture medium and repeatedly streak them on solid culture medium for purification. The purified strains are cultured at 25°C for 4 days. The pure colonies are then transferred to slant culture medium. After the colonies grow, they are stored in a refrigerator at 4°C. The culture is repeated every 2 weeks for initial screening.

[0026] Strain screening: The screened strains were inoculated into their corresponding liquid culture media, and after shaking culture for 72 h, the saccharifying enzyme activity in the fermentation broth was measured to identify the enzyme-producing strains.

[0027] (2) Colony morphology and strain identification

[0028] The isolated and purified strains were streaked onto LB solid medium plates and incubated at 25°C for 4 days. The morphology, size, color, and edge elevation of the colonies were observed. Single colonies were picked and Gram-stained for observation of colony morphology under a microscope.

[0029] The morphological and physiological-biochemical identification of this strain was performed according to the *Handbook of Fungal Identification*. Morphological identification and biological characteristic studies showed that after culturing *C. pekinensis* in LB solid medium at 25°C for 4 days, the colonies were milky white, large and thick, with a smooth, moist, and viscous surface. The colonies were uniform in texture and color, and the cells appeared oval under a microscope. Colony and cell morphology are as follows. Figure 1 As shown.

[0030] 16S rDNA gene sequencing was selected, and the sequencing results were compared with sequences in the NCBI database using BLAST homology search. The results showed that the CLYB1 strain sequence was similar to the published sequence of *Corynebacterium tumefaciens*. Clavispora lusitaniae 99% homology, phylogenetic tree as follows Figure 2 As shown.

[0031] Example 2: Determination of enzyme production capacity of the strain

[0032] The selected strains were shaken in the corresponding culture medium to activate them into bacterial suspensions, and the saccharifying enzyme activity of the bacterial suspensions was measured.

[0033] Prepare a 2% soluble starch solution, pH=4.6, 0.1 mol / L acetate buffer, 0.1 mol / L iodine solution, 0.1 mol / L NaOH solution, 2 mol / L sulfuric acid solution, 0.1 mol / L sodium thiosulfate solution, and 0.5% starch indicator.

[0034] Pipette 5 mL of 2% soluble starch solution and 2.5 mL of acetate buffer into a stoppered test tube, mix well, and incubate at 40°C for 10 min. Add 0.5 mL of bacterial suspension (the blank group is a boil-inactivated bacterial suspension), react at 40°C for 10 min, and inactivate in a boiling water bath for 5 min after the reaction. Take 5 mL of the reaction solution into an Erlenmeyer flask, add 5 mL of 0.1 mol / L iodine solution and 5 mL of 0.1 mol / L NaOH solution, shake well, and let stand in the dark for 15 min. Add 2 mL of 2 mol / L sulfuric acid solution and 1 mL of 0.5% starch indicator, and titrate with 0.1 mol / L sodium thiosulfate solution, recording the consumption. Calculate the glucoamylase activity using the following formula:

[0035]

[0036] In the formula, X represents the saccharifying enzyme activity (U / mL), V1 represents the sodium thiosulfate solution consumed in the blank group (mL), V2 represents the sodium thiosulfate solution consumed in the test group (mL), n represents the sodium thiosulfate concentration, 180.1 represents the molar mass of glucose, 8 / 5 represents 5 mL of the 8 mL reaction system, and 2 represents the specified 1 mL, but 0.5 mL was actually used.

[0037] Experimental results: The saccharifying enzyme activity in the fermentation broth of *Corynebacterium tumefaciens* was 73.19 ± 0.09 U / mL.

[0038] Example 3: Changes in soluble dietary fiber (SDF) content in rice bran obtained through homogenization and fermentation

[0039] Portuguese yeast was prepared using conventional methods. Clavispora lusitaniae CYPJ1, Kluyveromyces maculipena ( Kluyveromyces marxianus SXJ2, Bacillus sicca ( Siamese Bacillus DL-RXL01 bacterial solution, adjust the bacterial content in the solution to 1 x 10⁻⁶. 8 cfu / mL.

[0040] Homogenization process conditions: water to rice bran liquid ratio 19:1 (mL:g), homogenization speed 18000 rpm, homogenization time 6 min. After processing the rice bran, centrifuge at 3000 rpm for 10 min, collect the supernatant, add 4 volumes of anhydrous ethanol preheated at 60℃, allow to stand for precipitation for 4 h, centrifuge at 3000 rpm for 10 min, discard the supernatant, dry the bottom sediment at 40℃, then dissolve in water, and finally freeze-dry to obtain the SDF sample. The SDF yield calculation formula is as follows:

[0041] ;

[0042] In the formula: m SDF mass, g;M The mass of rice bran is expressed in grams.

[0043] Portuguese yeast ( Clavispora lusitaniae Fermentation conditions for CYPJ1: rice bran 50 g / L, starch 30 g / L, beef extract 20 g / L, inoculum 2% of fermentation liquid volume, fermentation time 72 h, fermentation temperature 28℃, fermentation pH 5.5. After fermenting rice bran, centrifuge at 3000 rpm for 10 min, collect the supernatant, add 4 times the volume of anhydrous ethanol preheated at 60℃, let stand for precipitation for 4 h, centrifuge at 3000 rpm for 10 min, discard the supernatant, dry the bottom sediment at 40℃, then dissolve in water, and freeze-dry to obtain SDF.

[0044] Homogenization aid for Portuguese Corynebacterium ( Clavispora lusitaniae The CYPJ1 fermentation process conditions were as follows: water to rice bran liquid ratio 19:1 (mL:g), homogenization speed 18000 rpm, homogenization time 6 min, starch 30 g / L, beef extract 20 g / L, inoculum 2% of fermentation liquid volume, fermentation time 72 h, fermentation temperature 28℃, fermentation pH 5.5. After fermenting the rice bran, the mixture was centrifuged at 3000 rpm for 10 min, the supernatant was collected, 4 times the volume of anhydrous ethanol preheated at 60℃ was added, the mixture was allowed to stand for precipitation for 4 h, centrifuged at 3000 rpm for 10 min, the supernatant was discarded, the bottom sediment was dried at 40℃, then dissolved in water, and then freeze-dried to obtain SDF.

[0045] Homogenization-assisted Max Kluyveromycin ( Kluyveromyces marxianus The SXJ2 fermentation process conditions were as follows: water to rice bran liquid ratio 19:1 (mL:g), homogenization speed 18000 rpm, homogenization time 6 min, starch 30 g / L, beef extract 20 g / L, inoculum amount 2% of fermentation liquid volume, fermentation time 72 h, fermentation temperature 28℃, fermentation pH 5.5. After fermenting the rice bran, the mixture was centrifuged at 3000 rpm for 10 min, the supernatant was collected, 4 times the volume of anhydrous ethanol preheated at 60℃ was added, the mixture was allowed to stand for precipitation for 4 h, centrifuged at 3000 rpm for 10 min, the supernatant was discarded, the bottom sediment was dried at 40℃, then dissolved in water, and then freeze-dried to obtain SDF.

[0046] Homogenized Bacillus simonii ( Siamese BacillusThe fermentation process conditions for DL-RXL01 were as follows: water to rice bran liquid ratio 19:1 (mL:g), homogenization speed 18000 rpm, homogenization time 6 min, starch 30 g / L, beef extract 20 g / L, inoculum amount 2% of fermentation liquid volume, fermentation time 72 h, fermentation temperature 28℃, fermentation pH 5.5. After fermenting the rice bran, the mixture was centrifuged at 3000 rpm for 10 min, the supernatant was collected, 4 times the volume of anhydrous ethanol preheated at 60℃ was added, the mixture was allowed to stand for 4 h to precipitate, centrifuged at 3000 rpm for 10 min, the supernatant was discarded, the bottom sediment was dried at 40℃, then dissolved in water, and then freeze-dried to obtain SDF.

[0047] Homogenization aid for Portuguese Corynebacterium ( Clavispora lusitaniae CYPJ1 and Kluyveromyces maculae ( Kluyveromyces marxianus SXJ2 fermentation process conditions: water to rice bran liquid-to-material ratio 19:1 (mL:g), homogenization speed 18000 rpm, homogenization time 6 min, starch added 30 g / L, beef extract 20 g / L, and Portuguese yeast ( Clavispora lusitaniae The inoculum size for CYPJ1 is 1% of the fermentation broth volume, containing Kluyveromyces macrocephala (…). Kluyveromyces marxianus The inoculum size of SXJ2 was 1% of the fermentation liquid volume, the fermentation time was 72 h, the fermentation temperature was 28℃, and the fermentation pH was 5.5. After fermenting rice bran, the mixture was centrifuged at 3000 rpm for 10 min, the supernatant was collected, 4 times the volume of anhydrous ethanol preheated at 60℃ was added, and the mixture was allowed to stand for precipitation for 4 h. After centrifugation at 3000 rpm for 10 min, the supernatant was discarded, the bottom sediment was dried at 40℃, then dissolved in water, and then freeze-dried to obtain SDF.

[0048] Homogenization aid for Portuguese Corynebacterium ( Clavispora lusitaniae CYPJ1 and Bacillus sicca ( Siamese Bacillus DL-RXL01 fermentation process conditions: water to rice bran liquid-to-material ratio 19:1 (mL:g), homogenization speed 18000 rpm, homogenization time 6 min, starch added 30 g / L, beef extract 20 g / L, and Portuguese yeast ( Clavispora lusitaniae The inoculum size for CYPJ1 was 1% of the fermentation broth volume, containing Bacillus sicca ( Bacillus Siamese The inoculum size of DL-RXL01 was 1% of the fermentation liquid volume, the fermentation time was 72 h, the fermentation temperature was 28℃, and the fermentation pH was 5.5. After fermenting rice bran, the mixture was centrifuged at 3000 rpm for 10 min, the supernatant was collected, 4 times the volume of anhydrous ethanol preheated at 60℃ was added, and the mixture was allowed to stand for precipitation for 4 h. After centrifugation at 3000 rpm for 10 min, the supernatant was discarded, the bottom sediment was dried at 40℃, then dissolved in water, and then freeze-dried to obtain SDF.

[0049] Each group underwent an average of three measurements, and the average value was taken. Statistical analysis was performed using SPSS 24.0 software. Quantitative data were expressed as x̅±s (mean ± standard deviation). The Kolmogorov-Smirnov test was used to test the normality of the data. For normally distributed data, the t-test was used to compare the differences in means between two groups, with P < 0.05 considered statistically significant. Specific results are shown in Table 1.

[0050] Table 1. Soluble dietary fiber (SDF) content in rice bran obtained by homogenization and fermentation

[0051]

[0052] Note: * indicates P<0.05 compared with homogenized fermentation + *C. pekinensis* and *Kluyveromyces martensii*, # indicates P<0.05 compared with homogenized fermentation + *C. pekinensis* and *Bacillus sicca*.

[0053] Example 4: Determination of relevant performance indicators of the prepared rice bran soluble dietary fiber (SDF)

[0054] The soluble dietary fiber (SDF) from rice bran prepared in Example 3 was tested for the following indicators:

[0055] 1. Water-holding capacity measurement

[0056] Accurately weigh 1.000 g (m1) of rice bran dietary fiber into a 50 mL beaker, add 20 mL of distilled water, and stir magnetically at room temperature for 4 h. Transfer the contents of the beaker to a centrifuge tube, centrifuge at 4000 r for 10 min, discard the supernatant, and weigh the wet weight of the precipitate (m2).

[0057] .

[0058] 2. Oil holding capacity measurement

[0059] Accurately weigh 1.000 g (m1) of rice bran dietary fiber and place it in a 50 mL beaker. Add 20 mL of vegetable oil and stir magnetically for 4 h. Transfer the contents of the beaker to a centrifuge tube and centrifuge at 8000 r for 10 min. Discard the supernatant and weigh the wet weight of the precipitate (m2).

[0060] .

[0061] 3. Measurement of water absorption swelling force

[0062] Accurately weigh 3.000 g of rice bran dietary fiber and place it in a 100 mL graduated cylinder. Measure the initial volume and record it as V1. Add 20 mL of distilled water, shake well, and let it stand at room temperature for 12 h. Measure the volume of the expanded rice bran dietary fiber and record it as V2.

[0063] .

[0064] 4. Determination of nitrite ion adsorption capacity

[0065] Preparation of the standard curve: Pipette 0.00, 0.20, 0.40, 0.60, 0.80, 1.00, 1.50, and 2.00 mL of 5 μg / mL NaNO₂ solution into test tubes, add 2 mL of 0.4% p-aminobenzenesulfonic acid, vortex to mix, let stand for 10 min, add 1 mL of 0.2% naphthylethylenediamine hydrochloride solution, and bring the volume to 10 mL with distilled water. Vortex to mix. Plot the absorbance at 538 nm with distilled water as the control on the ordinate and the nitrite ion concentration on the abscissa to construct the standard curve.

[0066] Sample determination: Accurately weigh 0.500 g of rice bran dietary fiber and place it in a 50 mL beaker. Add 20 mL of 200 μg / mL NaNO2 solution and adjust the pH to 2.0 and 7.0 respectively (simulating gastric juice and intestinal environment). Incubate at 37℃ for 2 h. After centrifugation, collect the supernatant and take 1 mL of the supernatant to determine the NaNO2 content.

[0067] Nitrite ion adsorption capacity = (NO2 content before adsorption - NO2 content after adsorption) / sample mass.

[0068] 5. Determination of glucose adsorption capacity

[0069] Preparation of standard curve: Pipette 1 mg / mL glucose standard solution into 20 mL test tubes, add distilled water to make up to 3 mL, add 2 mL DNS reagent, boil in water for 3 min, remove and cool to room temperature under running water, add 5 mL distilled water, shake to mix, and plot the standard curve with the absorbance measured at 520 nm as the ordinate and the glucose concentration as the abscissa, using distilled water as the control.

[0070] Sample determination: Accurately weigh 0.500 g of rice bran dietary fiber and place it in a 50 mL beaker. Add 20 mL of 1 mg / mL glucose solution and adjust the pH to 2.0 and 7.0 respectively (simulating gastric juice and intestinal environment). Incubate in a 37℃ water bath with shaking for 2 h. After centrifugation, collect the supernatant and determine the glucose content.

[0071] Glucose adsorption capacity = (glucose content before adsorption - glucose content after adsorption) / sample mass.

[0072] 6. Cholesterol Adsorption Capacity

[0073] Preparation of the standard curve: Pipette 0.00, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, and 0.40 mL of 0.1 mg / mL cholesterol solution into test tubes, respectively. Add glacial acetic acid to each test tube to a final volume of 0.4 mL. Then, add 0.4 mL of distilled water, 0.2 mL of 1 mg / mL phthalaldehyde reagent, and 4 mL of mixed acid (90% acetic acid and an equal volume of sulfuric acid). Shake well and let stand for 20 min. Plot the standard curve with the absorbance of the solution at 550 nm as the ordinate and the cholesterol content as the abscissa.

[0074] Sample determination: Fresh egg yolks were mixed with distilled water at a ratio of 1:9, and the absorbance was measured at 550 nm. The cholesterol content was observed to see if it was on the standard curve. If not, further dilution was performed. 1.0 g of rice bran dietary fiber was accurately weighed and placed in a 50 mL beaker. 25 mL of the diluted egg yolk emulsion was added, and the pH was adjusted to 2.0 and 7.0 respectively. The mixture was magnetically stirred for 4 h, transferred to centrifuge tubes, centrifuged at 4500 r for 10 min, and the supernatant was collected to determine its cholesterol content.

[0075] Cholesterol adsorption capacity = (cholesterol content before adsorption - cholesterol content after adsorption) / sample mass.

[0076] Each group of experiments was measured an average of three times, and the average value was taken. Statistical analysis was performed using SPSS 24.0 software. Quantitative data were expressed as x̅±s (mean ± standard deviation). The Kolmogorov-Smirnov test was used to test the normality of the data. For normally distributed data, the t-test was used to compare the differences in means between two groups. P<0.05 was considered statistically significant, and P<0.01 was considered extremely statistically significant. The specific results of various indicators of the prepared rice bran dietary fiber are shown in Tables 2-3.

[0077] Table 2. Results of water-holding capacity, oil-holding capacity, water absorption and swelling capacity, and nitrite ion adsorption capacity of the prepared rice bran soluble dietary fiber (SDF).

[0078]

[0079] Note: * indicates P<0.05 compared with homogenized + CYPJ1 and SXJ2 fermentation, # indicates P<0.05 compared with homogenized + CYPJ1 and DL-RXL01 fermentation.

[0080] Table 3. Results of glucose and cholesterol adsorption capacity determination of prepared rice bran soluble dietary fiber (SDF).

[0081]

[0082] Note: * indicates P<0.05 compared with homogenized + CYPJ1 and SXJ2 fermentation, # indicates P<0.05 compared with homogenized + CYPJ1 and DL-RXL01 fermentation.

[0083] Example 5: Mice were administered rice bran soluble dietary fiber (SDF) obtained through homogenization and fermentation by gavage.

[0084] Take the rice bran soluble dietary fiber (SDF) prepared in Example 3, wherein the rice bran soluble dietary fiber (SDF) is homogenized and assisted by *Corynebacterium tumefaciens* (Portunus spp.). Clavispora lusitaniae ) CYPJ1 fermentation process preparation.

[0085] The experimental animals were four-week-old male C57 / BL6 mice with an initial weight of 12.21 ± 0.22 g. They were randomly divided into four groups. The mice were acclimatized to a basal diet for one week, followed by a control group on the basal diet and the other groups on a high-fat diet. After 12 weeks of feeding, an intervention experiment was conducted. Once the hyperlipidemia mouse model was successfully established, the hyperlipidemia model group was randomly divided into three subgroups: HM group (hyperlipidemia model group), HM + LSDF group (hyperlipidemia model + low-dose SDF group, gavage dose of 0.5 g / kg / d), and HM + HSDF group (hyperlipidemia model + high-dose SDF group, gavage dose of 1.0 g / kg / d). The experiment lasted for 10 weeks, and mouse weight changes were recorded weekly. After the experiment, six mice from each group were randomly selected for bioinformatics analysis to evaluate the effects of different treatments on relevant mouse indicators.

[0086] 1. Mouse body weight

[0087] After gavage administration of rice bran dietary fiber, the weight gain trend of mice in the HM + LSDF (low-dose group) and HM + HSDF (high-dose group) groups was significantly slowed. At the end of gavage, compared with the NC (blank control) group, the HM (high-fat model) group showed a significant weight increase of 27.87%, while the HM + HSDF and HM + LSDF groups only increased by 12.65% and 15.00%, respectively. This indicates that dietary fiber SDF can effectively inhibit weight gain induced by a high-fat diet in mice, and the effect of high-dose SDF is more significant. Figure 3 ).

[0088] 2. Improved blood lipids in mice

[0089] Compared with the HM group, the levels of TC, TG, and LDL-C in the HM + LSDF and HM + HSDF groups decreased by 18.82%, 27.16%, and 23.51%, and 29.67%, 38.39%, and 50.52%, respectively; while the HDL-C levels in the HM + LSDF and HM + HSDF groups increased by 35.76% and 47.59%, respectively, with the HM + HSDF group showing the most significant improvement (P < 0.01). Rice bran dietary fiber intervention effectively regulated serum lipid levels in hyperlipidemic mice, significantly reducing TC, TG, and LDL-C levels while increasing HDL-C levels, thereby improving lipid metabolism disorders. Figure 4 ).

[0090] 3. Improvement of liver lipids in mice

[0091] Compared with the HM group, the levels of TC, TG, and LDL-C in the HM + LSDF and HM + HSDF groups decreased by 18.89%, 23.15%, and 11.73%, and 34.76%, 42.87%, and 26.27%, respectively. The levels of HDL-C in the HM + LSDF and HM + HSDF groups increased by 27.25% and 49.49%, respectively, with the HM + HSDF group showing the most significant improvement (P < 0.01). Rice bran SDF intervention effectively regulated liver lipid levels in hyperlipidemic mice, significantly reducing TC, TG, and LDL-C levels while increasing HDL-C levels, thereby improving lipid metabolism disorders. Figure 5 ).

[0092] 4. Improved liver function in mice

[0093] Compared with the HM group, the activities of ALT and AST in both the HM + LSDF group and the HM + HSDF group decreased by 17.46%, 22.95%, and 29.70%, 36.71%, respectively. The effect was most significant in the HM + HSDF group (P < 0.01), indicating that gavage administration of SDF, especially high-dose SDF, can effectively protect the liver of hyperlipidemic mice and reduce cell damage. Figure 6 ).

[0094] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A type of Portuguese yeast, characterized in that, The classification name of the Portuguese cork yeast is Portuguese cork yeast ( Clavispora lusitaniae CYPJ1, with accession number CGMCC No.35719.

2. The Portuguese yeast strain described in claim 1 ( Clavispora lusitaniae Application of CYPJ1 in glycan-producing enzymes.

3. The Portuguese yeast strain described in claim 1 ( Clavispora lusitaniae Application of CYPJ1 in the preparation of dietary fiber.

4. A compound microbial agent, characterized in that, The compound microbial agent includes *Corydalis lucida* (… Clavispora lusitaniae CYPJ1 and Kluyveromyces maculae ( Kluyveromyces marxianus SXJ2; or Portuguese yeast ( Clavispora lusitaniae CYPJ1 and Bacillus sicca ( Bacillus siamensis DL-RXL01; The Portuguese yeast ( Clavispora lusitaniae The CYPJ1 accession number is CGMCC No. 35719; the *Kluyveromyces martensii* strain (…) Kluyveromyces marxianus The SXJ2 accession number is CGMCC No. 14274; the Bacillus sicca described is... Bacillus siamensis The accession number for DL-RXL01 is CGMCC No. 23532.

5. The application of the compound microbial agent according to claim 4 in the preparation of dietary fiber.

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