A yeast phospholipid and a method for preparing the same

Abstract

The present application belongs to the technical field of yeast extract, and particularly relates to a yeast phospholipid and a preparation method thereof. The preparation method of the yeast phospholipid comprises the following steps: (1) dispersing yeast raw materials in an alcohol solution, then performing solid-liquid separation to obtain an extraction solution; (2) concentrating the extraction solution, then dispersing the extraction solution in an extraction solvent to perform extraction, separation, and concentrating a heavy phase solution to obtain the yeast phospholipid. In terms of the total mass of the yeast phospholipid, the crude fat contains 10% to 40% of phosphatidylcholine, 10% to 30% of phosphatidylethanolamine, and 1% to 10% of phosphatidylinositol. The preparation method of the present application extracts yeast raw materials by using a single solvent, is convenient for recovery, reduces the extraction cost, has higher extraction efficiency of the obtained phospholipid, has higher contents of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol, has higher content of unsaturated fatty acids, and is suitable for industrialized production.

Description

Technical Field

[0001] This invention belongs to the field of yeast extract technology, specifically relating to a yeast phospholipid and its preparation method. Background Technology

[0002] Phospholipids are lipid compounds containing phospholipid groups and are fundamental substances for life. Cell membranes are composed of approximately 40% protein and 50% lipids (mainly phospholipids). They are composed of lecithin, inositol phospholipids, and cephalin, among others. These phospholipids play corresponding functions in different parts and organs of the body. Phospholipids play a significant role in activating cells, maintaining metabolism, basal metabolism, and the balanced secretion of hormones, as well as enhancing the body's immunity and regenerative capacity. In addition, phospholipids also promote fat metabolism, prevent fatty liver, lower serum cholesterol, improve blood circulation, and prevent cardiovascular diseases.

[0003] Phospholipids are present in almost all body cells, and are abundant in important tissues of both plants and animals. Animal phospholipids mainly originate from egg yolks, milk, animal brain tissue, liver, kidneys, and muscle tissue. Plant phospholipids are mainly found in oilseeds, and most exist in the colloidal phase, bound to proteins, carbohydrates, fatty acids, phytosterols, vitamins, and other substances, making them an important byproduct of oil extraction. During oil extraction, phospholipids are extracted along with the oil. Crude soybean oil has the highest phospholipid content, making it the most important source of plant phospholipids. The soybean phospholipid extraction process is as follows: 1. Add 3% water to crude soybean oil and stir thoroughly at 60–80°C for 30 minutes. The phospholipids hydrate into a colloidal precipitate, which is then continuously centrifuged to obtain hydrated phospholipids. The precipitate is decolorized at 70°C with 3% hydrogen peroxide (1.5%). Then, it is dried under reduced pressure at 80–100°C and 2.67–8.00 kPa to obtain liquid phospholipids with a purity of 60%–70%. 2. Dissolve the oil and fatty acids in the phospholipids with 3 to 5 times the amount of acetone at 50°C, centrifuge to separate them, repeat the process twice, and finally dry under reduced pressure at 60°C to obtain phospholipid powder with a content of more than 95%.

[0004] Yeast is a single-celled microorganism belonging to the fungi class of higher microorganisms. It consists of a nucleus, cell membrane, cell wall, and mitochondria. It is rich in nutrients, widely used, and has a huge production capacity, with a global annual production of nearly 2 million tons. Its cell wall contains 16%–27% protein, 28%–30% mannan oligosaccharides, 25%–37% β-glucan, and 10%–20% fat. Mannan, β-glucan, and protein, which are the most abundant components of the cell wall, have all been extracted and utilized, but there are few reports on research related to the extraction of yeast phospholipids. Summary of the Invention

[0005] The technical problem solved by this invention is that there is limited research on yeast phospholipids and their extraction process in the prior art.

[0006] To address the above-mentioned problems, this invention provides a yeast phospholipid and its preparation method. The yeast phospholipid preparation method of this invention enriches the sources of phospholipids, and the yeast phospholipids obtained by this method have high contents of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol.

[0007] Specifically, the present invention provides the following technical solution:

[0008] In a first aspect, the present invention provides a yeast phospholipid, wherein the yeast phospholipid contains 20% to 100% crude fat based on the total mass of the yeast phospholipid; the crude fat is a lipid substance containing fatty acids;

[0009] The crude fat contains phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol; based on the total mass of yeast phospholipids, the crude fat contains 10%–40% phosphatidylcholine, 10%–30% phosphatidylethanolamine, and 1%–10% phosphatidylinositol.

[0010] In some embodiments, the yeast phospholipids contain 80% to 100% crude fat based on the total mass of the yeast phospholipids.

[0011] In some embodiments, the crude fat contains 20%–37% phosphatidylcholine, 17%–23% phosphatidylethanolamine, and 2%–5% phosphatidylinositol, based on the total mass of yeast phospholipids.

[0012] In some embodiments, the crude fat contains 30%–37% phosphatidylcholine, 20%–23% phosphatidylethanolamine, and 3%–5% phosphatidylinositol, based on the total mass of yeast phospholipids.

[0013] In some embodiments, the crude fat contains 35%–37% phosphatidylcholine, 20%–23% phosphatidylethanolamine, and 3%–5% phosphatidylinositol, based on the total mass of yeast phospholipids.

[0014] In some embodiments, the yeast phospholipids contain fatty acids with a carbon chain length of 16 carbon atoms, fatty acids with a carbon chain length of 17 carbon atoms, fatty acids with a carbon chain length of 18 carbon atoms, and fatty acids with a carbon chain length of 22 carbon atoms.

[0015] In some embodiments, the mass ratio of fatty acids with a carbon chain length of 16 carbon atoms, 17 carbon atoms, 18 carbon atoms, and 22 carbon atoms in the yeast phospholipid is (45-85):1:(50-80):(1-5).

[0016] In some implementations, the mass ratio of unsaturated fatty acids to saturated fatty acids is (1–5):1.

[0017] In some implementations, the mass ratio of unsaturated fatty acids to saturated fatty acids is (4-5):1.

[0018] Secondly, the present invention provides a method for preparing the yeast phospholipids, comprising the following steps:

[0019] (1) Disperse the yeast raw material in an alcohol solution, and then perform solid-liquid separation to obtain the extract;

[0020] (2) The extract obtained in step (1) is concentrated, then dispersed in an extraction solvent for extraction, separated, and the heavy phase solution is concentrated to obtain yeast phospholipids.

[0021] In some embodiments, the yeast feedstock is selected from yeast cells and / or yeast cell walls.

[0022] In some embodiments, the yeast includes one or more of Saccharomyces cerevisiae, Candida albicans, Gastrodia elata, Rhodotorula rubra, and Cryptococcus pluvialis.

[0023] In some embodiments, the yeast cell wall includes one or more of the following: Saccharomyces cerevisiae cell wall, Candida albicans cell wall, Gastrodia elata cell wall, Rhodotorula rubra cell wall, and Cryptococcus pluvialis cell wall.

[0024] In some implementations, in step (1), the mass of the added alcohol solution is 5 to 20 times the mass of the yeast raw material.

[0025] In some implementations, in step (1), the mass of the added alcohol solution is 9 to 15 times the mass of the yeast raw material.

[0026] In some embodiments, in step (1), the concentration of the alcohol solution is 50% to 100%.

[0027] In some embodiments, in step (1), the concentration of the alcohol solution is 60% to 90%.

[0028] In some embodiments, in step (1), the alcohol solution is selected from methanol solution, ethanol solution, n-propanol solution, isopropanol solution, n-butanol solution or n-pentanol solution.

[0029] In some implementations, the extraction temperature in step (1) is 25–105°C.

[0030] In some implementations, the extraction temperature in step (1) is 50–100°C.

[0031] In some implementations, the extraction temperature in step (1) is 50–75°C.

[0032] In some implementations, the extraction pressure in step (1) is 0.1 to 0.4 MPa.

[0033] In some implementations, the extraction time in step (1) is 2 to 15 hours.

[0034] In some implementations, the extraction time in step (1) is 10 to 15 hours.

[0035] In some implementations, in step (2), the volume of the extract is concentrated to 1 / 5 to 1 / 2 of the original extract volume.

[0036] In some embodiments, in step (2), the extraction solvent is selected from diethyl ether, petroleum ether, ethyl acetate, n-hexane, methyl tert-butyl ether, toluene, cyclohexane, or butyl acetate.

[0037] In some implementations, in step (2), the volume ratio of the extraction solvent to the concentrated extract is (0.5 to 1.5):1.

[0038] In some implementations, in step (2), the volume ratio of the extraction solvent to the concentrated extract is (0.8 to 1.2):1.

[0039] In some implementations, step (2) further includes decolorizing the heavy phase solution with activated carbon, filtering and separating it, and then concentrating the filtrate to obtain yeast phospholipids.

[0040] In some embodiments, step (2) further includes decolorizing the heavy phase solution with activated carbon, wherein the amount of activated carbon is 1 to 10 g; and / or, the decolorization temperature is 40-70°C; and / or, the decolorization time is 0.1 to 2 h.

[0041] In some embodiments, in step (2), the molar percentage of phosphorus in the heavy phase solution is 80% to 97%.

[0042] In some embodiments, when the yeast raw material is yeast, the yield of yeast phospholipids is 2% to 4%, and the content of acetone-insoluble matter is >60%.

[0043] In some embodiments, when the yeast raw material is yeast cell wall, the yield of yeast phospholipids is 8% to 12%, and the content of acetone-insoluble matter is >60%.

[0044] In some embodiments, when the yeast raw material is yeast, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol is 40% to 50% based on the total mass of yeast phospholipids.

[0045] In some embodiments, when the yeast raw material is a yeast cell wall, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol is 40% to 65% based on the total mass of yeast phospholipids.

[0046] In some embodiments, when the yeast feedstock is a yeast cell wall, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol is 60% to 65%.

[0047] In some embodiments, a yeast phospholipid is prepared by the method described above.

[0048] The beneficial effects of this invention are:

[0049] (1) The method for preparing yeast phospholipids in this invention is simple to operate and suitable for industrial production.

[0050] (2) In the process of extracting yeast phospholipids in this invention, no other components of yeast are damaged. The defatted yeast or cell wall can be reused, thus realizing the comprehensive utilization of resources.

[0051] (3) In the preparation process of yeast phospholipids in this invention, yeast cells or yeast cell walls are extracted using a single solvent, which simplifies the operation, facilitates recycling, and reduces extraction costs.

[0052] (4) In the preparation process of yeast phospholipids of the present invention, by using an extraction method, that is, concentrating the extract to a certain volume and then adding a specific extractant for extraction, the extraction efficiency of the obtained phospholipids is higher, the purity of the prepared phospholipids is higher, and the content of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol is higher.

[0053] (5) The yeast phospholipids prepared by this invention have a higher content of unsaturated fatty acids than saturated fatty acids, making them more valuable for application. Attached Figure Description

[0054] Figure 1 This is a flowchart of the yeast phospholipid extraction process. Detailed Implementation

[0055] As described above, the purpose of this invention is to provide a yeast phospholipid and a method for preparing the same.

[0056] In a first aspect, the present invention provides a yeast phospholipid, wherein, based on the total mass of the yeast phospholipid, the yeast phospholipid contains 20% to 100% crude fat, and the crude fat includes all lipid substances containing fatty acids, including phospholipids. Specifically, the crude fat contains phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol; based on the total mass of the yeast phospholipid, the crude fat contains 10% to 40% phosphatidylcholine, 10% to 30% phosphatidylethanolamine, and 1% to 10% phosphatidylinositol.

[0057] Preferably, the yeast phospholipids also contain phosphatidylserine and / or phosphatidylglycerol.

[0058] Preferably, the yeast phospholipid contains fatty acids with a carbon chain length of 16 carbon atoms, fatty acids with a carbon chain length of 17 carbon atoms, fatty acids with a carbon chain length of 18 carbon atoms, and fatty acids with a carbon chain length of 22 carbon atoms; the fatty acids are fatty acids in the yeast phospholipid structure or are free fatty acids.

[0059] Preferably, in the yeast phospholipids, the ratio of fatty acids with a carbon chain length of 16 carbon atoms, 17 carbon atoms, 18 carbon atoms, and 22 carbon atoms is (45-85):1:(50-80):(1-5).

[0060] Preferably, the ratio of unsaturated fatty acids to saturated fatty acids in the yeast phospholipids is (1-5):1.

[0061] Secondly, the present invention provides a method for preparing yeast phospholipids, comprising the following steps:

[0062] (1) Yeast raw material (yeast cells and / or yeast cell walls) is dispersed in a low alcohol solution for extraction. The extraction conditions are as follows: the extraction temperature is 25-105℃; and / or the extraction pressure is 0.1-0.4 MPa; and / or the extraction time is 2-15 h. The mass of the added alcohol solution is 9-15 times the mass of the yeast raw material, and the volume percentage of the alcohol solution is 50%-100%. Then, solid-liquid separation is performed. The filter cake is dried under reduced pressure at 80℃ and then recovered for other uses. The extract is kept for later use.

[0063] (2) The extract obtained in step (1) is concentrated to 1 / 5 to 1 / 2 of the original extract volume. Then it is dispersed in the extraction solvent for extraction. The volume ratio of the extraction solvent to the concentrated extract is (0.5 to 1.5):1. After separation, the heavy phase solution is decolorized with activated carbon, filtered and separated, and then the filtrate is concentrated to obtain yeast phospholipids.

[0064] The pressure extracted in this invention is the standard pressure, that is, a pressure value of 0.1 MPa refers to normal pressure.

[0065] In some embodiments, when the yeast raw material is yeast, the yield of yeast phospholipids is 2% to 4%, and the content of acetone-insoluble matter is >60%.

[0066] In some embodiments, when the yeast raw material is yeast cell wall, the yield of yeast phospholipids is 8% to 12%, and the content of acetone-insoluble matter is >60%.

[0067] In some embodiments, when the yeast raw material is yeast, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol is 40% to 50% based on the total mass of yeast phospholipids.

[0068] In some embodiments, when the yeast raw material is a yeast cell wall, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol is 40% to 65% based on the total mass of yeast phospholipids.

[0069] In some embodiments, when the yeast feedstock is a yeast cell wall, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol is 60% to 65%.

[0070] definition:

[0071] The yeast phospholipids described in this invention refer to products prepared from yeast or yeast cell walls, with phospholipids as the main component.

[0072] The fatty acids described in this invention mainly refer to the substituents on the phospholipid structure, and also contain a small amount of free fatty acids. Among them, fatty acids with a carbon chain length of 16 carbon atoms contain saturated fatty acids and unsaturated fatty acids; similarly, fatty acids with carbon chain lengths of 17, 18 and 22 carbon atoms respectively contain saturated fatty acids and unsaturated fatty acids.

[0073] Unless otherwise stated, all reagents / instruments used in the embodiments and comparative examples of this invention are conventional commercially available products / instruments. Information on the experimental materials and instruments used in this invention is shown in the table below:

[0074] Table 1 Experimental Materials / Instruments and Their Procurement Sources

[0075]

[0076]

[0077] To better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0078] Example 1

[0079] Add 5 kg of 80% ethanol to 1 kg of highly active dry yeast, heat to 100℃ with stirring, maintain the temperature at 0.2 MPa for 5 hours, cool to 40℃, filter, and dry the filter cake under reduced pressure at 80℃ for other uses. Concentrate 6 L of filtrate under reduced pressure to 3 L, add 3 L of petroleum ether, stir evenly, let stand for at least 30 minutes, and separate the layers. The light phase is 2.3 L, and the heavy phase is 3.7 L. Analyze the phosphorus content (calculated as P2O5) in both phases, as shown in Table 2. Add 3.7 g of activated carbon to the heavy phase, maintain the temperature at 50℃~60℃ with stirring for decolorization for 30 minutes, filter, and concentrate the filtrate under reduced pressure until no fraction flows out, yielding 25.4 g of paste-like yeast phospholipids. Based on the total mass of the highly active dry yeast raw material, the yield of yeast phospholipids is 2.54%.

[0080] Example 2

[0081] 10 kg of 90% methanol was added to 1 kg of highly active dry yeast. The mixture was heated to 70°C with stirring and kept at 0.1 MPa for 10 hours. The temperature was then lowered to 40°C, filtered, and the filter cake was dried under reduced pressure at 80°C for reuse. 12 L of the filtrate was concentrated under reduced pressure to 3 L, 3 L of n-hexane was added, and the mixture was stirred until homogeneous. After standing for at least 30 minutes to allow for complete separation, the liquid was separated into 2.3 L of light phase and 3.7 L of heavy phase. The phosphorus content (calculated as P2O5) in both phases was determined, as shown in Table 2. 3.7 g of activated carbon was added to the heavy phase, and the mixture was kept at 50°C–60°C with stirring for decolorization for 30 minutes. After filtration and concentration under reduced pressure, no distillate was effluent, yielding 32.1 g of paste-like yeast phospholipids. Based on the total mass of the highly active dry yeast raw material, the yield of yeast phospholipids was 3.21%.

[0082] Example 3

[0083] 8 kg of 70% isopropanol was added to 1 kg of Saccharomyces cerevisiae cell wall. The mixture was heated to 80°C with stirring and kept at 0.1 MPa for 8 hours. The temperature was then lowered to 40°C, filtered, and the filter cake was dried under reduced pressure at 80°C for reuse. 9 L of filtrate was concentrated under reduced pressure to 3 L, 3 L of ethyl acetate was added, and the mixture was stirred until homogeneous. After standing for at least 30 minutes to allow for complete separation, the light phase (2.4 L) and heavy phase (3.6 L) were separated. The phosphorus content (calculated as P2O5) in both phases was determined, as shown in Table 2. 3.6 g of activated carbon was added to the heavy phase, and the mixture was kept at 50°C–60°C with stirring for decolorization for 30 minutes. After filtration and concentration under reduced pressure, no distillate was effluent, yielding 85.6 g of paste-like yeast phospholipids. Based on the total mass of the Saccharomyces cerevisiae cell wall raw material, the yield of yeast phospholipids was 8.56%.

[0084] Example 4

[0085] 10 kg of 90% ethanol was added to 1 kg of Saccharomyces cerevisiae cell wall, and the mixture was heated to 60 °C with stirring. The mixture was then kept at 0.1 MPa and stirred for 15 hours, cooled to 40 °C, filtered, and the filter cake was dried under reduced pressure at 80 °C for reuse. 12 L of the filtrate was concentrated under reduced pressure to 3 L, and 3 L of methyl tert-butyl ether was added. The mixture was stirred evenly and allowed to stand for at least 30 minutes to allow for complete separation. The light phase was separated into 2.2 L and the heavy phase into 3.8 L. The phosphorus content (calculated as P2O5) in both phases was determined, as shown in Table 2. 3.8 g of activated carbon was added to the heavy phase, and the mixture was kept at 50 °C–60 °C with stirring for decolorization for 30 minutes. The mixture was filtered, and the concentration under reduced pressure yielded 98.3 g of paste-like yeast phospholipids. Based on the total mass of the Saccharomyces cerevisiae cell wall raw material, the yield of yeast phospholipids was 9.83%.

[0086] Example 5

[0087] 10 kg of 90% ethanol was added to 1 kg of Saccharomyces cerevisiae cell wall. The mixture was heated to 60 °C with stirring and kept at 0.1 MPa for 15 hours. The temperature was then lowered to 40 °C, filtered, and the filter cake was dried under reduced pressure at 80 °C for reuse. 12 L of the filtrate was concentrated to 3 L under reduced pressure, and 2 L of methyl tert-butyl ether was added. The mixture was stirred thoroughly and allowed to stand for at least 30 minutes to allow for complete separation. The light phase was separated into 1.2 L and the heavy phase into 3.8 L. The phosphorus content (calculated as P2O5) in both phases was determined, as shown in Table 2. 3.8 g of activated carbon was added to the heavy phase, and the mixture was kept at 50 °C–60 °C with stirring for decolorization for 30 minutes. After filtration and concentration under reduced pressure (no distillate flowed out), 102.8 g of yeast phospholipids were obtained. Based on the total mass of the Saccharomyces cerevisiae cell wall raw material, the yield of yeast phospholipids was 10.28%.

[0088] Example 6

[0089] 10 kg of 90% ethanol was added to 1 kg of Saccharomyces cerevisiae cell wall. The mixture was heated to 60 °C with stirring and kept at 0.1 MPa for 15 hours. The temperature was then lowered to 40 °C, filtered, and the filter cake was dried under reduced pressure at 80 °C for reuse. 12 L of the filtrate was concentrated to 3 L under reduced pressure, and 4.5 L of methyl tert-butyl ether was added. The mixture was stirred thoroughly and allowed to stand for at least 30 minutes to allow for complete separation. The light phase was separated into 4.2 L and the heavy phase into 3.3 L. The phosphorus content (calculated as P2O5) in both phases was determined, as shown in Table 2. 3.3 g of activated carbon was added to the heavy phase, and the mixture was kept at 50 °C–60 °C with stirring for decolorization for 30 minutes. After filtration and concentration under reduced pressure (no distillate flowed out), 87.2 g of yeast phospholipids were obtained. Based on the total mass of the Saccharomyces cerevisiae cell wall raw material, the yield of yeast phospholipids was 8.72%.

[0090] Comparative Example 1

[0091] 10 kg of 40% ethanol was added to 1 kg of Saccharomyces cerevisiae cell wall. The mixture was heated to 60 °C with stirring and kept at 0.1 MPa for 15 hours. The temperature was then lowered to 40 °C, filtered, and the filter cake was dried under reduced pressure at 80 °C for other uses. 12 L of filtrate was concentrated under reduced pressure to 3 L, 3 L of methyl tert-butyl ether was added, stirred evenly, and allowed to stand for at least 30 minutes to allow for complete separation. The light phase was separated into 3.2 L and the heavy phase into 2.8 L. The phosphorus content (calculated as P2O5) in the light and heavy phases was determined, as shown in Table 2. 2.8 g of activated carbon was added to the heavy phase, and the mixture was kept at 50 °C–60 °C with stirring for decolorization for 30 minutes. After filtration, the mixture was concentrated under reduced pressure until no distillate flowed out, yielding 32.3 g of yeast phospholipids. Based on the total mass of the Saccharomyces cerevisiae cell wall raw material, the yield of yeast phospholipids was 3.23%.

[0092] Comparative Example 2

[0093] Add 10 kg of 90% ethanol to the cell wall of 1 kg of Saccharomyces cerevisiae, heat to 60°C with stirring, maintain the temperature at 0.1 MPa for 15 hours with stirring, cool to 40°C, filter, and dry the filter cake under reduced pressure at 80°C for other uses. Take 2 L of the filtrate from 12 L of the filtrate, add 2 L of methyl tert-butyl ether, stir well, and let stand for more than 30 minutes until no separation occurs.

[0094] Comparative Example 3

[0095] 10 kg of 90% ethanol was added to 1 kg of Saccharomyces cerevisiae cell wall. The mixture was heated to 60 °C with stirring and kept at 0.1 MPa for 15 hours. The temperature was then lowered to 40 °C, filtered, and the filter cake was dried under reduced pressure at 80 °C for reuse. 12 L of the filtrate was concentrated to 1 L under reduced pressure, and 1 L of methyl tert-butyl ether was added. The mixture was stirred thoroughly and allowed to stand for at least 30 minutes to allow for complete separation. The light phase (0.8 L) and heavy phase (1.2 L) were separated, and the phosphorus content (calculated as P2O5) in both phases was determined, as shown in Table 2. 1.2 g of activated carbon was added to the heavy phase, and the mixture was kept at 50 °C–60 °C with stirring for decolorization for 30 minutes. After filtration and concentration under reduced pressure (no distillate flowed out), 72.8 g of yeast phospholipids were obtained. Based on the total mass of the Saccharomyces cerevisiae cell wall raw material, the yield of yeast phospholipids was 7.28%.

[0096] In the examples and comparative examples, the concentrated extract was added to the extractant and separated into light and heavy phases. The phosphorus content in the light and heavy phases was then measured. The instruments used for phosphorus content determination were an LC-08 digester (manufacturer: Shanghai Lichen Bangxi Instruments) and an SP-756PC UV-Vis spectrophotometer (manufacturer: Shanghai Spectrum Instruments Co., Ltd.). The phosphorus content detection method followed the People's Republic of China National Standard GB 5009.87-2016, National Food Safety Standard, Determination of Phosphorus in Food, Method I: Molybdenum Blue Spectrophotometric Method. The phosphorus content results obtained from the examples and comparative examples are shown in Table 2 below.

[0097] Table 2. Detection results of phosphorus (P2O5) in the light and heavy phases.

[0098]

[0099]

[0100] Fatty acids in the yeast phospholipid mixtures obtained in Examples 1-6 and Comparative Examples 1-3 were detected using a gas chromatography-mass spectrometry (GC-MS) system. The system consisted of an Agilent 6890 GC and an Agilent 5977 quadrupole mass spectrometer with a CP-Sil 88 (100m × 0.25mm × 0.25μm) column. GC analysis conditions were: injection volume 1 μL, split ratio 10:1, carrier gas high-purity helium, flow rate 1.0 mL / min; mass spectrometry conditions were: initial temperature 100℃ held for 5.0 min, temperature programmed to 240℃ at 5℃ / min, held for 10 min, injection port temperature 260℃, quadrupole temperature 150℃, single-channel scanning (SIM), scan range (m / z): 50–550. The proportions of several fatty acids with higher content in the yeast phospholipid mixtures, as well as the proportions of saturated and unsaturated fatty acids, are shown in Table 3.

[0101] Table 3. Results of the detection of major fatty acids in phospholipids

[0102]

[0103]

[0104] The contents of each phospholipid and other physicochemical indicators in the yeast phospholipid mixtures obtained in Examples 1-6 and Comparative Examples 1-3 were tested. The physicochemical indicators were tested in accordance with the People's Republic of China Grain Industry Standard LS / T3219-2017 Soybean Phospholipids. The test results of each phospholipid content and other physicochemical indicators are shown in Table 4.

[0105] Table 4. Results of Phospholipid Physicochemical Indicators

[0106]

[0107] While the specific embodiments of the present invention have been described above, they are not intended to limit the scope of protection of the present invention. Based on the technical solutions of the present invention, various modifications or variations that can be made by those skilled in the art without creative effort are still within the scope of protection of the present invention.

Claims

1. A method for preparing a yeast phospholipid, characterized by, The method comprises the following steps: (1) dispersing yeast and / or yeast cell wall in an alcohol solution to extract, and then performing solid-liquid separation to obtain an extract; The alcohol solution is selected from a methanol solution, an ethanol solution, a n-propanol solution, an isopropanol solution, a n-butanol solution or a n-pentanol solution; the volume percentage of the alcohol solution is 50% to 100%; the mass of the added alcohol solution is 5 to 20 times the mass of the yeast and / or yeast cell wall; (2) concentrating the extract obtained in step (1) to concentrate the volume of the extract to 1 / 5 to 1 / 2 of the original extract volume, and then dispersing in an extraction solvent to extract, separate, and take the heavy phase solution to concentrate to obtain yeast phospholipid; The extraction solvent is selected from diethyl ether, petroleum ether, ethyl acetate, n-hexane, methyl tert-butyl ether, toluene, cyclohexane or butyl acetate; the volume ratio of the extraction solvent to the concentrated extract is 1 to 1.5:1; The yeast phospholipid contains 20% to 100% crude fat in terms of the total mass of the yeast phospholipid; the crude fat is a lipid substance containing fatty acids; the phosphatidylcholine is 10% to 40%, the phosphatidylethanolamine is 10% to 30%, and the phosphatidylinositol is 1% to 10% in terms of the total mass of the yeast phospholipid.

2. The method for preparing a yeast phospholipid according to claim 1, wherein, The yeast includes one or more than two of Saccharomyces cerevisiae, Candida, Torulopsis, Rhodotorula and Cryptococcus; And / or, the yeast cell wall includes one or more than two of Saccharomyces cerevisiae cell wall, Candida cell wall, Torulopsis cell wall, Rhodotorula cell wall and Cryptococcus cell wall.

3. The method of preparing a yeast phospholipid according to claim 1, wherein, In step (1), the mass of the added alcohol solution is 9 to 15 times the mass of the yeast raw material.

4. The method of preparing a yeast phospholipid according to claim 2, wherein, In step (1), the mass of the added alcohol solution is 9 to 15 times the mass of the yeast raw material.

5. The method of preparing a yeast phospholipid according to claim 1, wherein, In step (1), the volume percentage of the alcohol solution is 60% to 90%.

6. The method of preparing a yeast phospholipid according to claim 2, wherein, In step (1), the volume percentage of the alcohol solution is 60% to 90%.

7. The method of preparing a yeast phospholipid according to claim 3, wherein, In step (1), the volume percentage of the alcohol solution is 60% to 90%.

8. The method of making a yeast phospholipid according to claim 1, wherein, In step (1), the volume percentage of the alcohol solution is 60% to 90%. In step (1), the temperature of the extraction is 25 to 105°C; And / or, the pressure of the extraction is 0.1 to 0.4 Mpa; 9. The method of making a yeast phospholipid according to claim 2, wherein, And / or, the time of the extraction is 2 to 15 h. In step (1), the temperature of the extraction is 25 to 105°C; And / or, the pressure of the extraction is 0.1 to 0.4 Mpa; 10. The method of making a yeast phospholipid according to claim 3, wherein, And / or, the time of the extraction is 2 to 15 h. In step (1), the temperature of the extraction is 25 to 105°C; And / or, the pressure of the extraction is 0.1 to 0.4 Mpa; 11. The method of making a yeast phospholipid according to claim 5, wherein, And / or, the time of the extraction is 2 to 15 h. In step (1), the temperature of the extraction is 25 to 105°C; And / or, the pressure of the extraction is 0.1 to 0.4 Mpa; 12. The method of making a yeast phospholipid according to claim 8, wherein, And / or, the time of the extraction is 2 to 15 h.

13. The method of making a yeast phospholipid according to claim 8, wherein, In step (1), the temperature of the extraction is 25 to 105°C; 14. The method of making a yeast phospholipid according to claim 8, wherein, And / or, the pressure of the extraction is 0.1 to 0.4 Mpa; 15. The method of preparing a yeast phospholipid according to any one of claims 1-14, wherein, And / or, the time of the extraction is 2 to 15 h. In step (1), the temperature of the extraction is 50 to 100°C. In step (1), the temperature of the extraction is 50 to 75°C. In step (1), the time of the extraction is 10 to 15 h. In step (2), the volume ratio of the extraction solvent to the concentrated extract is 1 to 1.2:

1.

16. The method of preparing a yeast phospholipid according to any one of claims 1-14, wherein, In step (2), the heavy phase solution is further subjected to decolorization with activated carbon, separated by filtration, and then the obtained yeast phospholipid is obtained by concentrating the filtrate.

17. The method of making a yeast phospholipid according to claim 15, wherein, In step (2), the heavy phase solution is further subjected to decolorization with activated carbon, separated by filtration, and then the obtained yeast phospholipid is obtained by concentrating the filtrate.

18. The method of making a yeast phospholipid according to any one of claims 1-14, wherein, When the yeast raw material is yeast cells, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol is 40% to 50% based on the total mass of the yeast phospholipid; When the yeast raw material is yeast cells, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol is 40% to 50% based on the total mass of the yeast phospholipid; 19. The method of making a yeast phospholipid according to claim 15, wherein, When the yeast raw material is yeast cells, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol is 40% to 50% based on the total mass of the yeast phospholipid; When the yeast raw material is yeast cells, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol is 40% to 50% based on the total mass of the yeast phospholipid; 20. The method of making a yeast phospholipid according to claim 16, wherein, When the yeast raw material is yeast cells, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol is 40% to 50% based on the total mass of the yeast phospholipid; When the yeast raw material is yeast cells, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol is 40% to 50% based on the total mass of the yeast phospholipid; 21. The method of making a yeast phospholipid according to claim 18, wherein, When the yeast raw material is yeast cells, the total mass percentage of phosphatidylcholine, phosphatidylethanolamine and phosphatidylinositol is 40% to 50% based on the total mass of the yeast phospholipid;

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