Preparation method of vitamin B yeast extract, composition containing vitamin B yeast extract and application of vitamin B yeast extract
By using high-pressure synergistic enzymatic hydrolysis and high-pressure homogenization technology to break down yeast cells, the problem of B vitamin damage in the yeast extraction process was solved, thereby increasing the content of B vitamins and protein in the yeast extract. This has the effect of enhancing osteoblast function and inhibiting osteoporosis.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-02-02
- Publication Date
- 2026-03-17
AI Technical Summary
Existing yeast extraction processes often damage heat- and shear-sensitive B vitamins when disrupting the yeast cell wall, resulting in insufficient impact on their content and activity. Therefore, there is a lack of effective extraction methods.
A yeast extract rich in B vitamins was prepared by using a high-pressure synergistic enzymatic method. This involved preparing an active yeast suspension, enzymatic hydrolysis with complex enzymes such as β-1,3-glucanase, chitinase, and cellulase, followed by high-pressure homogenization, centrifugation to collect the supernatant after cell wall disruption, and drying.
It significantly increased the content of B vitamins and protein yield in yeast extract, enhanced its regenerative protective effect on osteoblasts, and effectively inhibited the development of osteoporosis.
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Figure CN121667390A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of food technology, specifically the field of sports nutrition food technology, and relates to a method for preparing a yeast extract of B vitamins, a composition containing the extract, and its application. Background Technology
[0002] Yeast extract (YE) is a natural food ingredient or biological raw material made from edible yeast through processes such as enzymatic hydrolysis, thermal reaction, separation, and purification. This process breaks down macromolecules such as proteins, nucleic acids, and carbohydrates within yeast cells into smaller nutrient molecules, which are then concentrated and dried. Morphologically, yeast extract is commonly found as a pale yellow to brownish-yellow powder, paste, or liquid. It has a rich meaty, umami, or savory aroma, good solubility, and no noticeable off-odor, making it an important natural flavor enhancer and nutritional fortifier in the food industry.
[0003] Yeast extract has the effect of increasing bone density and inhibiting osteoporosis, mainly through multiple mechanisms such as regulating bone metabolism balance, promoting osteoblast activity, inhibiting osteoclast proliferation and differentiation, and improving calcium absorption. From a physiological perspective, the active components in yeast extract, such as β-glucan, amino acids, vitamin D precursors, and minerals like calcium, phosphorus, and zinc, can synergistically act on bone metabolism pathways: On the one hand, β-glucan can promote osteoblast proliferation and differentiation by activating the PI3K / Akt signaling pathway, upregulating the expression of osteogenic-related genes such as osteocalcin and alkaline phosphatase, and accelerating bone matrix synthesis and mineralization; on the other hand, its active peptides can inhibit the expression of nuclear factor κB receptor activator ligand (RANKL), reducing osteoclast formation and activity, and lowering the rate of bone resorption, thereby maintaining the dynamic balance between bone formation and bone resorption. Furthermore, the vitamin D precursors in yeast extract can be converted into active vitamin D, promoting intestinal calcium absorption and increasing blood calcium concentration to provide raw materials for bone mineralization; minerals such as zinc and phosphorus participate in the formation of the bone matrix, enhancing the mechanical strength of bone tissue. Related studies have shown that long-term supplementation with yeast extract can significantly increase femoral bone mineral density and reduce the level of bone turnover markers in ovariectomized osteoporosis model rats, providing a natural and safe potential intervention for the clinical prevention and improvement of osteoporosis.
[0004] In the extraction process of yeast extract, due to the relatively tough yeast cell wall, disrupting the yeast cell wall is crucial for releasing yeast proteins. Methods mainly include mechanical, physical, chemical, and enzymatic methods. Among these, mechanical disruption technology is simple to operate, low in cost, and scalable. While destructive methods can efficiently release the contents, they can also easily damage heat-sensitive and shear-sensitive bioactive substances, such as B vitamins. Therefore, research on yeast extraction processes is essential for improving its quality and functionality. However, current research mainly focuses on increasing protein yield, lacking studies on the impact on B vitamin content and activity.
[0005] To obtain more effective, safe, natural yeast extracts rich in protein and B vitamins, optimizing extraction methods and treating bioactivity retention rate as a core indicator of equal importance to extraction rate is a future research direction. Summary of the Invention
[0006] This invention relates to a method for preparing a yeast extract containing B vitamins, a composition comprising the extract, and its applications. Specifically, the technical solution of this invention is implemented as follows: In a first aspect, the present invention provides a method for preparing a yeast extract containing B vitamins, characterized by comprising the following steps: S1. Prepare an active yeast suspension; S2. The active yeast suspension is hydrolyzed with a compound enzyme to obtain an active yeast hydrolysate; S3. The active yeast hydrolysate from step S2 is homogenized under high pressure, centrifuged, and the supernatant is dried to obtain the final product.
[0007] Preferably, step S1 includes the following steps: S101. Weigh the yeast, dissolve it in 1~10% (m / v) glucose solution, place it at 30~40℃ and keep it for 10~120 min to obtain an active yeast solution. S102. Centrifuge the active yeast solution, discard the supernatant, resuspend the centrifuged precipitate, adjust the cell density, and obtain an active yeast suspension.
[0008] Preferably, step S2 includes the following steps: S201, Prepare the compound enzyme; S202. Add the compound enzyme to the active yeast suspension prepared in step S1, place it at 40~60℃, and hydrolyze for 24~48h to obtain active yeast hydrolysate.
[0009] Preferably, step S3 includes the following steps: S301. Take the active yeast hydrolysate prepared in step S2, adjust the pH, and then place it in a high-pressure homogenizer for high-pressure homogenization. S302. After high-pressure homogenization, centrifuge, take the supernatant, dry it into powder, and you will get the product.
[0010] More preferably, in step S101, the yeast is selected from one or more of Candida utilis, Saccharomyces cerevisiae, Hansenula polymorpha, and Pichia pastoris.
[0011] More preferably, in step S102, the cell density is 1*102 6 Cells / mL ~ 1*10 9 Cells / mL.
[0012] More preferably, in step S201, the complex enzyme is selected from two or more of β-1,3-glucanase, chitinase, cellulase, pectinase, amylase, protease, and mannanase.
[0013] More preferably, in step S201, the complex enzyme is β-1,3-glucanase, chitinase, and cellulase.
[0014] Most preferably, in step S201, the complex enzyme is composed of β-1,3-glucanase, chitinase, and cellulase mixed in a mass ratio of (8~13):1:(1~3).
[0015] More preferably, in step S301, the conditions for high-pressure homogenization are: homogenization pressure 50~80MPa, homogenization times 2~4 times, and homogenization time per time 1~3min.
[0016] More preferably, in step S301, the pH is adjusted to 7-8.
[0017] More preferably, in step S302, the drying method is selected from air drying, radiation drying, spray drying, and freeze drying.
[0018] In a second aspect, the present invention provides a composition containing B vitamins and yeast extract.
[0019] The composition has a positive effect on the regeneration or protection of osteoblasts.
[0020] The composition has the effect of increasing bone density.
[0021] The composition inhibits the development of osteoporosis.
[0022] The composition is used to treat or improve osteoporosis.
[0023] The preparation method of the B vitamin yeast extract is as follows: S1. Prepare an active yeast suspension; S2. The active yeast suspension is hydrolyzed with a compound enzyme to obtain an active yeast hydrolysate; S3. The active yeast hydrolysate from step S2 is homogenized under high pressure, centrifuged, and the supernatant is dried to obtain the final product.
[0024] Preferably, step S1 includes the following steps: S101. Weigh the yeast, dissolve it in 1~10% (m / v) glucose solution, place it at 30~40℃ and keep it for 10~120 min to obtain an active yeast solution. S102. Centrifuge the active yeast solution, discard the supernatant, resuspend the centrifuged precipitate, adjust the cell density, and obtain an active yeast suspension.
[0025] Preferably, step S2 includes the following steps: S201, Prepare the compound enzyme; S202. Add the compound enzyme to the active yeast suspension prepared in step S1, place it at 40~60℃, and hydrolyze for 24~48h to obtain active yeast hydrolysate.
[0026] Preferably, step S3 includes the following steps: S301. Take the active yeast hydrolysate prepared in step S2, adjust the pH, and then place it in a high-pressure homogenizer for high-pressure homogenization. S302. After high-pressure homogenization, centrifuge, take the supernatant, dry it into powder, and you will get the product.
[0027] More preferably, in step S101, the yeast is selected from one or more of Candida utilis, Saccharomyces cerevisiae, Hansenula polymorpha, and Pichia pastoris.
[0028] More preferably, in step S102, the cell density is 1*102 6 Cells / mL ~ 1*10 9 Cells / mL.
[0029] More preferably, in step S201, the complex enzyme is selected from two or more of β-1,3-glucanase, chitinase, cellulase, pectinase, amylase, protease, and mannanase.
[0030] More preferably, in step S201, the complex enzyme is β-1,3-glucanase, chitinase, and cellulase.
[0031] Most preferably, in step S201, the complex enzyme is composed of β-1,3-glucanase, chitinase, and cellulase mixed in a mass ratio of (8~13):1:(1~3).
[0032] More preferably, in step S301, the conditions for high-pressure homogenization are: homogenization pressure 50~80MPa, homogenization times 2~4 times, and homogenization time per time 1~3min.
[0033] More preferably, in step S301, the pH is adjusted to 7-8.
[0034] More preferably, in step S302, the drying method is selected from air drying, radiation drying, spray drying, and freeze drying.
[0035] Thirdly, the present invention provides a method for preparing B vitamin yeast extract and its application in the preparation of food or pharmaceuticals.
[0036] This invention provides the use of a composition containing B vitamins and yeast extract in the preparation of food or pharmaceuticals.
[0037] Preferably, the food is selected from health food and sports nutrition food.
[0038] The food has a positive effect on the regeneration or protection of osteoblasts.
[0039] The food has the effect of increasing bone density.
[0040] The food can inhibit the development of osteoporosis.
[0041] The food product can be used to improve osteoporosis.
[0042] The beneficial effects of this invention are as follows: High-pressure synergistic enzymatic method is used to break down yeast cells, yielding a yeast extract rich in protein and B vitamins. Analysis of zebrafish calcification matrix shows that the yeast extract prepared by this invention significantly increases bone density in zebrafish and has a positive effect on osteoblast regeneration and protection. When applied to sports nutrition foods, it effectively inhibits the development of osteoporosis. Attached Figure Description
[0043] Figure 1 This is a morphological image of a zebrafish.
[0044] Figure 2 The image shows the fluorescence detection results of the zebrafish calcified matrix. Detailed Implementation
[0045] The present invention will be further described below with reference to specific embodiments, but the present invention is not limited to these embodiments. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.
[0046] The Candida utilis used in this invention is ATCC 22023.
[0047] β-1,3-glucanase, CAS 9044-93-3, purchased from MERCK.
[0048] Chitinase, CAS 9001-06-3, purchased from Hubei Langbowan Biomedical Co., Ltd.
[0049] Cellulase, CAS 9012-54-8, purchased from Beijing Bailingwei Technology Co., Ltd.
[0050] Yeast lyase (Zymolyase, Arthrobacter luteus) was purchased from MCE HY-E70306.
[0051] Dexamethasone, purchased from Sangon Biotech A601187-0005.
[0052] Calcein, purchased from Sangon Biotech, A602198-0025.
[0053] Example 1 A method for preparing a yeast extract containing B vitamins includes the following steps: S1. Preparation of active yeast suspension: S101. Weigh 8g of Candida utilis and dissolve it in 1L of 5% (m / v) glucose solution. Place it at 35℃ and keep it for 60min to obtain an active yeast solution. S102. Centrifuge the active yeast solution at 1000 rpm for 3 min, discard the supernatant, and resuspend the centrifuged precipitate in acetic acid solution at pH 5.5, adjusting the density to 1*10⁻⁶. 8 Cells / mL, to obtain an active yeast suspension; S2. The active yeast suspension is hydrolyzed using a compound enzyme to obtain an active yeast hydrolysate: S201. Preparation of compound enzyme: Mix β-1,3-glucanase, chitinase and cellulase in a mass ratio of 10:1:3 to obtain compound enzyme; S202. Add the compound enzyme to the active yeast suspension prepared in step S1 at a dosage of 0.8% (m / v), place it at 50℃, and enzymatically hydrolyze for 36 hours to obtain active yeast hydrolysate. S3. Homogenize the active yeast hydrolysate from step S2 under high pressure, centrifuge to collect the supernatant, and dry it. S301. Take the active yeast hydrolysate prepared in step S2, adjust the pH to 7 with 1 mol / L NaOH solution, and then place it in a high-pressure homogenizer for high-pressure homogenization. The homogenization pressure is 70 MPa, the homogenization is performed 3 times, and the homogenization time is 2 min per homogenization. S302. After high-pressure homogenization, centrifuge at 2000 rpm for 5 min, take the supernatant, freeze-dry to obtain powder, and you will get the product.
[0054] Example 2
[0055] A method for preparing a yeast extract containing B vitamins includes the following steps: S1. Preparation of active yeast suspension: S101. Weigh 10g of Candida utilis, dissolve it in 1L of 4% (m / v) glucose solution, place it at 37℃ for 30min to obtain an active yeast solution. S102. Centrifuge the active yeast solution at 1000 rpm for 3 min, discard the supernatant, and resuspend the centrifuged precipitate in acetic acid solution at pH 5.5, adjusting the density to 1*10⁻⁶. 8 Cells / mL, to obtain an active yeast suspension; S2. The active yeast suspension is hydrolyzed using a compound enzyme to obtain an active yeast hydrolysate: S201. Preparation of compound enzyme: Mix β-1,3-glucanase, chitinase and cellulase in a mass ratio of 12:1:2 to obtain compound enzyme; S202. Add the compound enzyme at a dosage of 1% (m / v) to the active yeast suspension prepared in step S1, place it at 45℃, and enzymatically hydrolyze for 40 h to obtain active yeast hydrolysate. S3. Homogenize the active yeast hydrolysate from step S2 under high pressure, centrifuge to collect the supernatant, and dry it. S301. Take the active yeast hydrolysate prepared in step S2, adjust the pH to 7 with 1 mol / L NaOH solution, and then place it in a high-pressure homogenizer for high-pressure homogenization. The homogenization pressure is 70 MPa, the homogenization is performed 3 times, and the homogenization time is 2 min per homogenization. S302. After high-pressure homogenization, centrifuge at 2000 rpm for 5 min, take the supernatant, freeze-dry to obtain powder, and you will get the product.
[0056] Example 3 A method for preparing a yeast extract containing B vitamins includes the following steps: S1. Preparation of active yeast suspension: S101. Weigh 12g of Candida utilis, dissolve it in 1L of 4% (m / v) glucose solution, place it at 33℃ and keep it for 40min to obtain an active yeast solution. S102. Centrifuge the active yeast solution at 1000 rpm for 3 min, discard the supernatant, and resuspend the centrifuged precipitate in acetic acid solution at pH 5.5, adjusting the density to 1*10⁻⁶. 8 Cells / mL, to obtain an active yeast suspension; S2. The active yeast suspension is hydrolyzed using a compound enzyme to obtain an active yeast hydrolysate: S201. Preparation of compound enzyme: Mix β-1,3-glucanase, chitinase and cellulase in a mass ratio of 9:1:2 to obtain compound enzyme; S202. Add the compound enzyme at a dosage of 1% (m / v) to the active yeast suspension prepared in step S1, place it at 55℃, and enzymatically hydrolyze for 30 h to obtain active yeast hydrolysate. S3. Homogenize the active yeast hydrolysate from step S2 under high pressure, centrifuge to collect the supernatant, and dry it. S301. Take the active yeast hydrolysate prepared in step S2, adjust the pH to 7 with 1 mol / L NaOH solution, and then place it in a high-pressure homogenizer for high-pressure homogenization. The homogenization pressure is 70 MPa, the homogenization is performed 3 times, and the homogenization time is 2 min per homogenization. S302. After high-pressure homogenization, centrifuge at 2000 rpm for 5 min, take the supernatant, freeze-dry to obtain powder, and you will get the product.
[0057] Comparative Example 1 A method for preparing a yeast extract containing B vitamins includes the following steps: S1. Preparation of active yeast suspension: S101. Weigh 8g of Candida utilis and dissolve it in 1L of 5% (m / v) glucose solution. Place it at 35℃ and keep it for 60min to obtain an active yeast solution. S102. Centrifuge the active yeast solution at 1000 rpm for 3 min, discard the supernatant, and resuspend the centrifuged precipitate in PBS buffer, adjusting the density to 1*10⁻⁶. 8 Cells / mL, to obtain an active yeast suspension; S2. The active yeast suspension from step S1 is homogenized under high pressure, centrifuged, and the supernatant is collected and dried. S201. Take the active yeast suspension prepared in step S1 and place it in a high-pressure homogenizer for high-pressure homogenization. The homogenization pressure is 140MPa, the number of homogenizations is 3, and the homogenization time is 3min per homogenization. S202, after high-pressure homogenization, centrifuge at 2000 rpm for 5 min, take the supernatant, freeze-dry to obtain powder, and thus obtain.
[0058] Comparative Example 2 A method for preparing a yeast extract containing B vitamins includes the following steps: S1. Preparation of active yeast suspension: S101. Weigh 8g of Candida utilis and dissolve it in 1L of 5% (m / v) glucose solution. Place it at 35℃ and keep it for 60min to obtain an active yeast solution. S102. Centrifuge the active yeast solution at 1000 rpm for 3 min, discard the supernatant, and resuspend the centrifuged precipitate in acetic acid solution at pH 5.5, adjusting the density to 1*10⁻⁶. 8 Cells / mL, to obtain an active yeast suspension; S2. The active yeast suspension is hydrolyzed using a compound enzyme to obtain an active yeast hydrolysate: S201. Preparation of compound enzyme: Mix β-1,3-glucanase, chitinase and cellulase in a mass ratio of 10:1:3 to obtain compound enzyme; S202. Add the compound enzyme to the active yeast suspension prepared in step S1 at a dosage of 0.8% (m / v), place it at 50℃, and enzymatically hydrolyze for 36 hours to obtain active yeast hydrolysate. S203, after enzymatic hydrolysis, adjust the pH to 7 with 1 mol / L NaOH solution, then centrifuge at 2000 rpm for 5 min, take the supernatant, freeze-dry to obtain powder, and thus obtain the product.
[0059] Comparative Example 3 A method for preparing a yeast extract containing B vitamins includes the following steps: S1. Preparation of active yeast suspension: S101. Weigh 10g of Candida utilis, dissolve it in 1L of 4% (m / v) glucose solution, place it at 37℃ for 30min to obtain an active yeast solution. S102. Centrifuge the active yeast solution at 1000 rpm for 3 min, discard the supernatant, and resuspend the centrifuged precipitate in acetic acid solution at pH 5.5, adjusting the density to 1*10⁻⁶. 8 Cells / mL, to obtain an active yeast suspension; S2. The active yeast suspension is hydrolyzed using a compound enzyme to obtain an active yeast hydrolysate: S201. Preparation of compound enzyme: Mix β-1,3-glucanase, chitinase and cellulase in a mass ratio of 3:2:2 to obtain compound enzyme; S202. Add the compound enzyme at a dosage of 1% (m / v) to the active yeast suspension prepared in step S1, place it at 45℃, and enzymatically hydrolyze for 40 h to obtain active yeast hydrolysate. S3. Homogenize the active yeast hydrolysate from step S2 under high pressure, centrifuge to collect the supernatant, and dry it. S301. Take the active yeast hydrolysate prepared in step S2, adjust the pH to 7 with 1 mol / L NaOH solution, and then place it in a high-pressure homogenizer for high-pressure homogenization. The homogenization pressure is 70 MPa, the homogenization is performed 3 times, and the homogenization time is 2 min per homogenization. S302. After high-pressure homogenization, centrifuge at 2000 rpm for 5 min, take the supernatant, freeze-dry to obtain powder, and you will get the product.
[0060] Comparative Example 4 A method for preparing a yeast extract containing B vitamins includes the following steps: S1. Preparation of active yeast suspension: S101. Weigh 10g of Candida utilis, dissolve it in 1L of 4% (m / v) glucose solution, place it at 37℃ for 30min to obtain an active yeast solution. S102. Centrifuge the active yeast solution at 1000 rpm for 3 min, discard the supernatant, and resuspend the centrifuged precipitate in acetic acid solution at pH 5.5, adjusting the density to 1*10⁻⁶. 8 Cells / mL, to obtain an active yeast suspension; S2. The active yeast suspension was hydrolyzed using β-1,3-glucanase to obtain the active yeast hydrolysate: S201. Add β-1,3-glucanase at a dosage of 1% (m / v) to the active yeast suspension prepared in step S1, place it at 45℃, and hydrolyze for 40 h to obtain active yeast hydrolysate. S3. Homogenize the active yeast hydrolysate from step S2 under high pressure, centrifuge to collect the supernatant, and dry it. S301. Take the active yeast hydrolysate prepared in step S2, adjust the pH to 7 with 1 mol / L NaOH solution, and then place it in a high-pressure homogenizer for high-pressure homogenization. The homogenization pressure is 70 MPa, the homogenization is performed 3 times, and the homogenization time is 2 min per homogenization. S302. After high-pressure homogenization, centrifuge at 2000 rpm for 5 min, take the supernatant, freeze-dry to obtain powder, and you will get the product.
[0061] Comparative Example 5 A method for preparing a yeast extract containing B vitamins includes the following steps: S1. Preparation of active yeast suspension: S101. Weigh 10g of Candida utilis, dissolve it in 1L of 4% (m / v) glucose solution, place it at 37℃ for 30min to obtain an active yeast solution. S102. Centrifuge the active yeast solution at 1000 rpm for 3 min, discard the supernatant, and resuspend the centrifuged precipitate in PBS buffer, adjusting the density to 1*10⁻⁶. 8 Cells / mL, to obtain an active yeast suspension; S2. The active yeast suspension was hydrolyzed with Zymolyase to obtain the active yeast hydrolysate: Zymolyase was added to the active yeast suspension prepared in step S1 at a concentration of 0.1 mg / mL, and the mixture was placed at 40°C for 40 h to obtain the active yeast hydrolysate. S3. Homogenize the active yeast hydrolysate from step S2 under high pressure, then centrifuge to collect the supernatant and dry it. S301. Take the active yeast hydrolysate prepared in step S2 and place it in a high-pressure homogenizer for high-pressure homogenization. The homogenization pressure is 70 MPa, the homogenization is performed 3 times, and the homogenization time is 2 min per time. S302. After high-pressure homogenization, centrifuge at 2000 rpm for 5 min, take the supernatant, freeze-dry to obtain powder, and you will get the product.
[0062] Test Example 1: Determination of B Vitamin Content in Yeast Extract Yeast extract powders prepared in Examples 1-3 and Comparative Examples 1-5 were used to determine the content of various B vitamins using the test methods in Table 1. The test sample was the only variable. The results are shown in Table 2.
[0063] Table 1. Methods for determining the content of B vitamins Vitamins Test methods B1 Thiamine GB 5009.84-2016 National Food Safety Standard: Determination of Vitamin B1 in Food B2 Riboflavin GB 5009.85-2016 National Food Safety Standard: Determination of Vitamin B2 in Food B3 Niacin / Niacinamide GB 5009.89-2023 National Food Safety Standard: Determination of Niacin and Nicotinamide in Food B5 pantothenic acid GB 5009.210-2023 National Food Safety Standard: Determination of Pantothenic Acid in Food B6 pyridoxine GB 5009.154-2023 National Food Safety Standard: Determination of Vitamin B6 in Food B9 Folic Acid GB 5009.211-2022 National Food Safety Standard: Determination of Folic Acid in Food
[0064] Table 2 Results of B vitamin content determination (unit: mg / 100g) B1 B2 B3 B5 B6 B9 Example 1 6.57 4.63 59.88 10.12 20.56 2.94 Example 2 6.64 4.35 56.75 10.16 22.68 3.15 Example 3 6.13 4.40 57.62 9.46 23.49 3.38 Comparative Example 1 1.72 1.94 41.79 6.67 15.33 1.40 Comparative Example 2 3.01 2.26 42.08 8.14 16.80 1.88 Comparative Example 3 3.58 3.15 45.48 8.60 16.41 1.97 Comparative Example 4 4.27 3.43 41.65 7.97 17.03 2.16 Comparative Example 5 4.69 3.20 44.40 8.15 17.89 2.43
[0065] The results showed that the preparation methods of Examples 1-3 of this invention preserved the B vitamins in yeast to the greatest extent, with excellent extraction effects, showing substantial differences compared to Comparative Examples 1-5. Comparative Example 1 used high-pressure homogenization to disrupt the cell wall, but even with increased homogenization pressure, it could not effectively extract B vitamins from yeast, indicating that the extraction efficiency of high-pressure homogenization alone is low. Comparative Example 2 used enzymatic hydrolysis to disrupt the cell wall, with slightly better results than high-pressure homogenization, but the extraction efficiency of B vitamins was still very limited, again demonstrating the low extraction efficiency of enzymatic hydrolysis alone. Comparative Examples 3, 4, and 5 adjusted the proportion and type of the compound enzymes, confirming that the type and amount of compound enzymes in the high-pressure synergistic enzymatic method of this invention have significant advantages in exerting core performance.
[0066] Test Example 2: Determination of Nitrogen Content in Yeast Extract
[0067] Yeast extract powders prepared in Examples 1-3 and Comparative Examples 1-5 were tested using the methods for total nitrogen (6.4) and amino acid nitrogen (6.5) in GB / T 23530-2009 "Yeast Extracts". The test sample was the only variable. The results are shown in Table 3.
[0068] Table 3 Results of amino acid nitrogen content determination Total nitrogen content (%) Amino acid nitrogen content (%) Example 1 10.7% 4.7% Example 2 11.0% 4.7% Example 3 11.2% 4.8% Comparative Example 1 8.7% 3.2% Comparative Example 2 9.3% 3.4% Comparative Example 3 9.6% 4.0% Comparative Example 4 9.5% 3.6% Comparative Example 5 9.8% 4.1%
[0069] The results showed that the total nitrogen content and amino acid nitrogen content of the yeast extracts in Examples 1-3 of this invention were significantly better than those in Comparative Examples 1-5. The high-pressure synergistic enzymatic method of this invention increases the content of B vitamins while maintaining a high protein yield.
[0070] Test Example 3: Efficacy Test of Yeast Extract Using zebrafish as a model organism for functional testing, the effect of the yeast extract powder prepared in Example 1 of this invention on improving osteoporosis was evaluated by using the green fluorescently labeled calcein-calcium ion fluorescent complex as a test index for improving osteoporosis.
[0071] Test methods 1. Wild-type male and female zebrafish of strain AB were cultured separately in water at 28℃. Culture conditions included a 14h / 10h light / dark cycle, with dissolved oxygen maintained at 6-8 mg / L. Live food was provided daily. Zebrafish were placed in a breeding tank at a female-to-male ratio of 2:1, separated by a partition. The partition was removed before the next day's light exposure to allow free fertilization. Fertilized eggs were collected and placed in clean E3 medium in a 28.5℃ incubator. Experiments began three days after fertilization.
[0072] 2. Set up a control group, a model group, and a sample group. The control group was cultured in culture water. The model group and the sample group were established by adding dexamethasone to the culture water. The sample group was given a certain amount of yeast extract powder prepared in Example 1 of this invention at the same time as adding dexamethasone.
[0073] 3. Calcein staining was performed on day 9 (9dpf), and images were acquired using a stereofluorescence microscope.
[0074] Test results: Morphological examination of zebrafish showed that, compared with the control group, no significant changes in zebrafish morphology were observed at concentrations of 1000 μg / mL, 2000 μg / mL, and 3000 μg / mL of the yeast extract powder of this invention. Teratogenicity and mortality were within the normal range. Figure 1 Finally, the median value of 2000 μg / mL was selected as the optimal test concentration.
[0075] Analysis of the fluorescence detection results of zebrafish calcified matrix ( Figure 2 The study revealed differences between the sample group and the model group. In the model group, zebrafish treated with dexamethasone showed significant impairment of osteoblast function, increased osteoclast function, and decreased bone density. Compared to the model group, the sample group exhibited significantly increased vertebral bone mineralization. In terms of vertebral bone fluorescence intensity, the sample group significantly increased bone density in zebrafish, demonstrating a positive effect on osteoblast regeneration and protection, and inhibiting the development of osteoporosis.
[0076] The basic principles, main features, and advantages of this invention have been described above. Those skilled in the art should understand that this invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely principles of the invention. Various changes and modifications can be made without departing from the spirit and scope of the invention, and all such changes and modifications fall within the scope of the invention as claimed. The scope of protection claimed by this invention is defined by the appended claims and their equivalents.
Claims
1. A method for preparing a B-vitamin yeast extract, characterized in that, It comprises the following steps: S1, preparing a live yeast suspension; S2, enzymatically hydrolyzing the live yeast suspension with a complex enzyme to obtain a live yeast hydrolysate; S3, homogenizing the live yeast hydrolysate of step S2 under high pressure, centrifuging to obtain the supernatant, and drying, to obtain the product.
2. The production method according to claim 1, wherein The step S1 comprises the following steps: S101, weighing the yeast, dissolving it in a 1-10% (m / v) glucose solution, and placing it at 30-40℃ for 10-120 min to obtain a live yeast solution; S102, centrifuging the live yeast solution, discarding the supernatant, resuspending the centrifugal precipitate, and adjusting the cell density to obtain a live yeast suspension.
3. The production method according to claim 1, wherein The step S2 comprises the following steps: S201, preparing a complex enzyme; S202, adding the complex enzyme to the live yeast suspension prepared in step S1, and placing it at 40-60℃ for 24-48 h to obtain a live yeast hydrolysate.
4. The production method according to claim 1, wherein The step S3 comprises the following steps: S301, taking the live yeast hydrolysate prepared in step S2, adjusting the pH, and then placing it in a high-pressure homogenizer for high-pressure homogenization; S302, centrifuging after high-pressure homogenization, taking the supernatant, and drying it into a powder, to obtain the product.
5. The production method according to claim 2, wherein In step S101, the yeast is selected from one or more of Candida utilis, Saccharomyces cerevisiae, Hansenula, and Pichia.
6. The production method according to claim 3, wherein In step S201, the complex enzyme is selected from two or more of β-1,3-glucanase, chitinase, cellulase, pectinase, amylase, protease, and mannanase.
7. The production method according to claim 4, wherein In step S301, the high-pressure homogenization is performed under the following conditions: homogenization pressure of 50-80 MPa, homogenization times of 2-4, and single homogenization time of 1-3 min.
8. A composition comprising a B-vitamin yeast extract, characterized in that, The B-vitamin-containing yeast extract is prepared by the method of claim 1.
9. The composition of claim 8, wherein The composition has a positive effect on the regeneration or protection of osteoblasts; the composition has the effect of increasing bone density; the composition inhibits the development of osteoporosis; and the composition is used for treating or improving osteoporosis.
10. Use of the B-vitamin-containing yeast extract composition of claim 8 in the preparation of food.