Method for co-producing ergosterol and yeast sterol from yeast

Abstract

The invention relates to the field of yeast extraction, in particular to a method for co-producing ergosterol and yeast sterol from yeast. The invention provides a method for co-producing ergosterol and yeast sterol from yeast, 4-phenyl-3H-1, 2, 4-triazoline-3, 5-diketone can selectively react with ergosterol in a sterol mixed solution, and the yeast sterol does not participate in the reaction, so that the purposes of removing the ergosterol and purifying the yeast sterol are achieved. The co-production of the yeast sterol is realized from the yeast for producing the ergosterol, and the economic value is improved while the tailing discharge is reduced. Compared with an existing technology for extracting yeast sterol from yeast, the technology does not need multi-step chromatographic separation, the process is greatly simplified, the selectivity is high, and the purity of yeast sterol is high.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of yeast extraction, in particular to a method for preparing ergosterol and zymosterol by co-production from yeast. BACKGROUND

[0002] Ergosterol has been prepared by yeast extraction for a long time, and it is also the current mainstream commercial production scheme of ergosterol. Zymosterol is the core intermediate of the biosynthetic pathway of sterol substances, and can be used as a substrate to synthesize ergosterol, chalinosterol, cholesterol and other sterol substances through enzymatic reaction. Zymosterol also has certain drug development potential, and can be used for research on antifungal targets and anticancer mechanisms.

[0003] In the metabolic process of yeast synthesizing ergosterol, not all of the accumulated zymosterol is converted, only a part of it is converted into ergosterol and other metabolites. Therefore, there are mainly two sterol substances in the yeast producing ergosterol: ergosterol and zymosterol. When yeast produces ergosterol, zymosterol as an impurity is removed in the purification process and is enriched in the mother liquor, and is finally treated as waste.

[0004] There are many technical schemes for extracting ergosterol from yeast, but there are few schemes for extracting zymosterol, and there is no scheme for recovering zymosterol from the mother liquor after extracting ergosterol. In the article Sterol synthesis: a simple method for the isolation of zymosterol (5 alpha-cholesta-8, 24-dien-3 beta-ol) from yeast and spectral properties of zymosterol, the technical scheme adopted is as follows: the yeast is refluxed in 15% potassium hydroxide ethanol solution for 12h, the reaction solution is poured into water, the organic phase is taken after petroleum ether extraction and water washing, column chromatography, fraction collection of eluate and esterification, and then column chromatography to obtain zymosterol ester, and finally hydrolysis to obtain zymosterol with high purity.

[0005] Since the yeast contains both zymosterol and ergosterol, but the solubility difference between the two is large, zymosterol as an impurity can be easily removed when ergosterol is extracted from yeast. However, if you want to extract zymosterol alone, it is difficult to remove ergosterol as an impurity. Therefore, in the above prior art scheme, several column chromatographies are needed, and esterification and hydrolysis reactions are also needed to obtain zymosterol. The process route is long, and several column chromatographies make the operation cumbersome. In addition, sterols can be esterified and hydrolyzed, so the selectivity will not be high. SUMMARY

[0006] In view of the above, the present application provides a method for preparing ergosterol and zymosterol by co-production of yeast. The method uses 4-phenyl-3H-1,2,4-triazoline-3,5-dione to selectively react with ergosterol in a mixed solution of sterols, while zymosterol does not participate in the reaction, thereby removing ergosterol and purifying zymosterol.

[0007] To achieve the above-mentioned purposes, the present application provides the following technical solutions:

[0008] In a first aspect, the present application provides a method for preparing zymosterol, comprising the following steps:

[0009] Step 1: obtaining a crystallization mother liquor after extracting ergosterol from yeast, concentrating, and collecting the concentrate;

[0010] Step 2: dissolving the concentrate in ethyl acetate, washing with water, separating the liquid, and collecting the organic phase;

[0011] Step 3: reacting the organic phase with 4-phenyl-3H-1,2,4-triazoline-3,5-dione, concentrating, dissolving in ethanol, first cooling and crystallizing, collecting the filtrate by suction filtration, second cooling and crystallizing, filtering, and drying to obtain the zymosterol; the filtering is suction filtration;

[0012] The crystallization in step 3 includes first crystallization and second crystallization; the temperature of the first crystallization is 20-25℃; and the temperature of the second crystallization is 0-10℃;

[0013] The amount of 4-phenyl-3H-1,2,4-triazoline-3,5-dione used in step 3 is 1.1-1.5 equivalents;

[0014] The purity of the 4-phenyl-3H-1,2,4-triazoline-3,5-dione is not less than 94%.

[0015] In some embodiments of the present application, the mass ratio of ergosterol in the crystallization mother liquor to the 4-phenyl-3H-1,2,4-triazoline-3,5-dione is 2.4: (1.16-1.59).

[0016] In some embodiments of the present application, the mass ratio of ergosterol in the crystallization mother liquor to the 4-phenyl-3H-1,2,4-triazoline-3,5-dione is 2.4:1.16, 2.4:1.38, or 2.4:1.59.

[0017] In some embodiments of the present application, the first concentration in step 1 is 50-60℃ concentration under reduced pressure.

[0018] In some embodiments of the present application, the temperature of the reaction in step 3 is 25℃±5℃, and the time of the reaction is 2h.

[0019] In some embodiments of the present application, the concentration in step 3 is concentration under reduced pressure at 40~50℃.

[0020] In some embodiments of the present application, the temperature of the dissolution in step 3 is 70~80℃, preferably 78℃.

[0021] In some embodiments of the present application, the temperature of the first cooling crystallization in step 3 is 20~25℃, preferably 20℃.

[0022] In some embodiments of the present application, the temperature of the second cooling crystallization in step 3 is 0~10℃, preferably 0℃.

[0023] In some embodiments of the present application, the preparation method of the crystallization mother liquor after the extraction of ergosterol from the yeast in step 1 comprises:

[0024] Step 1-1: taking the yeast to perform saponification reaction in alkali substance and water, performing solid-liquid separation, and collecting the first solid phase;

[0025] Step 1-2: adding a first solvent to the first solid phase to extract ergosterol, performing solid-liquid separation to obtain a second liquid phase and a second solid phase, and drying the second solid phase to obtain defatted yeast;

[0026] Step 1-3: concentrating the second liquid phase to dryness under reduced pressure, dissolving in a second solvent, washing with water, separating, collecting the organic layer, concentrating under reduced pressure, adding ethanol to crystallize to obtain ergosterol, and collecting the crystallization mother liquor, thereby obtaining the crystallization mother liquor after the extraction of ergosterol from the yeast;

[0027] In step 1-2, the first solvent comprises any one or more of ethanol, n-propanol, isopropanol, n-butanol, acetone, acetonitrile;

[0028] In step 1-3, the second solvent comprises any one or more of n-hexane, toluene, ethyl acetate, petroleum ether, cyclohexane, methyl tert-butyl ether, dichloromethane, chloroform.

[0029] In step 1-1, the alkali substance comprises one or more of potassium hydroxide, sodium hydroxide, lithium hydroxide, sodium ethoxide, and sodium methoxide aqueous solution;

[0030] In step 1-1, the mass ratio of the yeast, the alkali substance, and water is 1:(0.1~0.5):(5~15).

[0031] In step 1-1, the saponification reaction is performed for 4-12 hours; and the saponification reaction is performed at a temperature of 60-100°C.

[0032] In some embodiments of the present application, in step 1-1, the saponification reaction is performed at a temperature of 60-80°C.

[0033] In some embodiments of the present application, in step 1-2, the mass-volume ratio of the yeast to the first solvent is 1: (10-20) in kg / L.

[0034] In step 1-2, the extraction is performed at a temperature of 56-120°C for 1-3 hours.

[0035] In some embodiments of the present application, in step 1-2, the ratio of the yeast, the alkali substance and water is 1: (0.2-0.3): (10-13) by mass.

[0036] In some embodiments of the present application, in step 1-3, the second solvent is used in an amount of twice the volume of the yeast.

[0037] In step 1-3, the concentration under reduced pressure is performed at a temperature of 40-80°C.

[0038] In step 1-3, the crystallization is performed using the ethanol in a mass ratio of (0.5-1.5) to the yeast; and the crystallization is performed at a temperature of 0-30°C.

[0039] In some embodiments of the present application, the yeast includes one or more of Saccharomyces cerevisiae, Candida, Torulopsis, Rhodotorula or Cryptococcus.

[0040] In a second aspect, the present application also provides a method for preparing ergosterol and zymosterol by co-production from yeast, which comprises the following steps:

[0041] extracting ergosterol from the yeast;

[0042] collecting the crystallization mother liquor after the yeast extracts the ergosterol, and obtaining zymosterol according to the method for preparing zymosterol described above.

[0043] In some embodiments of the present application, the method for extracting ergosterol from the yeast comprises:

[0044] Step 1-1: subjecting the yeast to a saponification reaction in an alkali substance and water, performing solid-liquid separation, and collecting a first solid phase;

[0045] Step 1-2: adding a first solvent to the first solid phase to extract ergosterol, performing solid-liquid separation to obtain a second liquid phase and a second solid phase, and drying the second solid phase to obtain defatted yeast;

[0046] Step 1-3: The second liquid phase is concentrated to dryness under reduced pressure, a second solvent is added to dissolve, washed with water, separated, the organic layer is collected and concentrated under reduced pressure, ethanol is added to crystallize to obtain ergosterol;

[0047] In step 1-2, the first solvent includes any one or more of ethanol, n-propanol, isopropanol, n-butanol, acetone, acetonitrile;

[0048] In step 1-3, the second solvent includes any one or more of n-hexane, toluene, ethyl acetate, petroleum ether, cyclohexane, methyl tert-butyl ether, dichloromethane, chloroform.

[0049] In some specific embodiments of the present application, in step 1-1, the base substance includes one or more of potassium hydroxide, sodium hydroxide, lithium hydroxide, sodium ethoxide, and aqueous sodium methoxide;

[0050] In step 1-1, the mass ratio of the yeast, the base substance, and water is 1: (0.1-0.5): (5-15);

[0051] In step 1-1, the saponification reaction time is 4-12 hours; the saponification reaction temperature is 60-100℃;

[0052] In step 1-1, the saponification reaction temperature is 60-80℃;

[0053] In step 1-2, the mass-volume ratio of the yeast to the first solvent is 1: (10-20) in kg / L;

[0054] In step 1-2, the extraction temperature is 56-120℃, and the reflux time is 1-3 hours;

[0055] In step 1-2, the mass ratio of the yeast, the base substance, and water is 1: (0.2-0.3): (10-13);

[0056] In step 1-3, the amount of the second solvent is twice the volume of the yeast;

[0057] In step 1-3, the reduced pressure concentration temperature is 40-80℃;

[0058] In step 1-3, the mass ratio of the ethanol to the yeast used for crystallization is (0.5-1.5): the crystallization temperature is 0-30℃.

[0059] In some specific embodiments of the present application, the yeast includes one or more of Saccharomyces cerevisiae, Candida, Torulopsis, Rhodotorula, or Cryptococcus.

[0060] The present application provides a method for preparing ergosterol and zymosterol by coproduction of yeast, which uses 4-phenyl-3H-1,2,4-triazole-3,5-dione to selectively react with ergosterol in a mixed solution of sterols, while zymosterol does not participate in the reaction, so as to remove ergosterol and purify zymosterol.

[0061] The beneficial effects of the present application include but are not limited to:

[0062] (1) The coproduction of zymosterol is realized from the yeast producing ergosterol, which reduces the discharge of tailings and improves the economic value.

[0063] (2) Compared with the existing process for extracting zymosterol from yeast, the present process does not need multi-step chromatographic separation, greatly simplifies the process, and has high selectivity and high purity of zymosterol. BRIEF DESCRIPTION OF DRAWINGS

[0064] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings needed to be used in the embodiments or the prior art description will be briefly introduced below.

[0065] Figure 1 The ratio of zymosterol and ergosterol in the yeast extract solution of the prior art is shown;

[0066] Figure 2 The ratio of ergosterol and zymosterol in the ergosterol crystallization mother liquor of the prior art is shown;

[0067] Figure 3 The process flow diagram of the present application is shown;

[0068] Figure 4 The principle diagram of the present application is shown;

[0069] Figure 5 The HPLC spectrum of zymosterol in Example 1 is shown;

[0070] Figure 6 The HPLC spectrum of zymosterol in Example 2 is shown;

[0071] Figure 7 The HPLC spectrum of zymosterol in Example 3 is shown;

[0072] Figure 8 The HPLC spectrum of zymosterol in Example 4 is shown;

[0073] Figure 9 The HPLC spectrum of zymosterol in Comparative Example 1 is shown;

[0074] Figure 10 The HPLC spectrum of zymosterol in Comparative Example 2 is shown;

[0075] Figure 11 The HPLC spectrum of zymosterol in Comparative Example 3 is shown;

[0076] Figure 12 The HPLC overlay chromatograms of each sample and yeast sterol standard are shown. Detailed Implementation

[0077] This invention discloses a method for the co-production of ergosterol and yeast sterol from yeast. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired result. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can obviously make modifications or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0078] Technical solution flow of this invention:

[0079] (1) Referring to our authorized patent CN115960156B "A method for extracting ergosterol using yeast", the ergosterol extraction method is as follows: after the product is separated by crystallization, a crystallization mother liquor is obtained. The mother liquor is concentrated and the solvent is recovered. The concentrated liquid is dissolved in ethyl acetate, washed with water, and separated to obtain an ethyl acetate solution containing yeast sterol and ergosterol.

[0080] (2) Add an ethyl acetate solution of 4-phenyl-3H-1,2,4-triazolline-3,5-dione to the solution. 4-phenyl-3H-1,2,4-triazolline-3,5-dione reacts with ergosterol, while yeast sterol does not participate in the reaction.

[0081] (3) Concentrate to remove ethyl acetate, dissolve the residue with ethanol at elevated temperature, and then cool to 20-25°C to precipitate the solid after the ergosterol reaction.

[0082] (4) Filter to remove solids, and further cool the solution to 0-10℃ to crystallize. After filtration and drying, yeast sterol is obtained.

[0083] This invention uses the amount of 4-phenyl-3H-1,2,4-triazoline-3,5-dione and the crystallization temperature of yeast sterols as variables. The former mainly affects the purity of yeast sterols, while the latter mainly affects the yield. The amount of 4-phenyl-3H-1,2,4-triazoline-3,5-dione is preferably 1.1-1.5 equivalents (purity above 94%). Too much will result in waste, while too little will lead to incomplete ergosterol reaction and reduce the purity of the yeast sterol sample. The crystallization temperature is preferably below 0-10℃, where the yield does not change significantly. Too low a temperature wastes energy, while too high a temperature will lead to a decrease in yield.

[0084] The raw materials and reagents used in the method for preparing ergosterol and zymosterol by co-production of yeast are commercially available.

[0085] The application will be further described below in combination with examples:

[0086] Example 1:

[0087] Take 2L ergosterol crystallization mother liquor (containing 2.4g of ergosterol and 6.2g of zymosterol, respectively), 50-60℃, reduced pressure concentration into solid, add 1L ethyl acetate to dissolve, then add 2L water to stir for 10min, static separation, take the organic phase, add 50g anhydrous magnesium sulfate to dry, drop 20ml of 1.16g (1.1eq) 4-phenyl-3H-1,2,4-triazoline-3,5-dione dissolved in ethyl acetate solution at room temperature under stirring, react for 2h, 40-50℃, reduced pressure concentration to oil, add 100ml ethanol to dissolve at reflux, reduce to 25℃ to crystallize, filter, further reduce the filtrate to 0℃ to crystallize, filter, dry to obtain 5.62g of zymosterol, purity 94.5%, as shown in Figure 5 .

[0088] Example 2:

[0089] Take 2L ergosterol crystallization mother liquor (containing 2.4g of ergosterol and 6.2g of zymosterol, respectively), 50-60℃, reduced pressure concentration into solid, add 1L ethyl acetate to dissolve, then add 2L water to stir for 10min, static separation, take the organic phase, add 50g anhydrous magnesium sulfate to dry, drop 20ml of 1.38g (1.3eq) 4-phenyl-3H-1,2,4-triazoline-3,5-dione dissolved in ethyl acetate solution at room temperature under stirring, react for 2h, 40-50℃, reduced pressure concentration to oil, add 100ml ethanol to dissolve at reflux, reduce to 20℃ to crystallize, filter, further reduce the filtrate to 0℃ to crystallize, filter, dry to obtain 5.59g of zymosterol, purity 94.8%, as shown in Figure 6 .

[0090] Example 3:

[0091] Take 2 L ergosterol crystallization mother liquor (containing ergosterol 2.4 g and yeast sterol 6.2 g, respectively), 50-60 ℃ reduced pressure concentration into solid, add 1 L ethyl acetate dissolution, add 2 L water stirring 10 min, static separation, take the organic phase, add 50 g anhydrous magnesium sulfate drying, drop 20 ml dissolved in 1.59 g (1.5 eq) 4-phenyl-3H-1,2,4-triazole-3,5-diketone ethyl acetate solution under stirring at room temperature, reaction 2 h, 40-50 ℃ reduced pressure concentration to oil, add 100 ml ethanol to reflux dissolution, drop to 23 ℃ crystallization, filtration, further drop to 5 ℃ crystallization, filtration, drying to get 5.53 g yeast sterol, purity 95.2%, as shown in Figure 7 .

[0092] Example 4:

[0093] Take 2 L ergosterol crystallization mother liquor (containing ergosterol 2.4 g and yeast sterol 6.2 g, respectively), 50-60 ℃ reduced pressure concentration into solid, add 1 L ethyl acetate dissolution, add 2 L water stirring 10 min, static separation, take the organic phase, add 50 g anhydrous magnesium sulfate drying, drop 20 ml dissolved in 1.38 g (1.3 eq) 4-phenyl-3H-1,2,4-triazole-3,5-diketone ethyl acetate solution under stirring at room temperature, reaction 2 h, 40-50 ℃ reduced pressure concentration to oil, add 100 ml ethanol to reflux dissolution, drop to 25 ℃ crystallization, filtration, further drop to 10 ℃ crystallization, filtration, drying to get 5.49 g yeast sterol, purity 95.0%, as shown in Figure 8 .

[0094] Comparative Example 1:

[0095] Take 2 L ergosterol crystallization mother liquor (containing ergosterol 2.4 g and yeast sterol 6.2 g, respectively), 50-60 ℃ reduced pressure concentration into solid, add 1 L ethyl acetate dissolution, add 2 L water stirring 10 min, static separation, take the organic phase, add 50 g anhydrous magnesium sulfate drying, drop 20 ml dissolved in 1.06 g (1.0 eq) 4-phenyl-3H-1,2,4-triazole-3,5-diketone ethyl acetate solution under stirring at room temperature, reaction 2 h, 40-50 ℃ reduced pressure concentration to oil, add 100 ml ethanol to reflux dissolution, drop to 20 ℃ crystallization, filtration, further drop to 5 ℃ crystallization, filtration, drying to get 5.73 g yeast sterol, purity 92.6%, as shown in Figure 9 .

[0096] Comparative Example 2:

[0097] Take 2L ergosterol crystallization mother liquor (containing ergosterol 2.4g and yeast sterol 6.2g respectively), 50-60℃ reduced pressure concentration into solid, add 1L ethyl acetate dissolution, add 2L water stirring 10min, static liquid, take the organic phase, add 50g anhydrous magnesium sulfate drying, drop 20ml dissolved in 1.69g (1.6eq) 4-phenyl-3H-1,2,4-triazole-3,5-diketone ethyl acetate solution under stirring at room temperature, reaction 2h, 40-50℃ reduced pressure concentration to oil, add 100ml ethanol to reflux dissolution, drop to 20℃ crystallization, filtration, further drop to 15℃ crystallization, filtration, drying to get 4.64g yeast sterol, purity 95.2%, as shown in Figure 10 .

[0098] Comparative example 3:

[0099] Take 2L ergosterol crystallization mother liquor (containing ergosterol 2.4g and yeast sterol 6.2g respectively), 50-60℃ reduced pressure concentration into solid, add 1L ethyl acetate dissolution, add 2L water stirring 10min, static liquid, take the organic phase, add 50g anhydrous magnesium sulfate drying, drop 20ml dissolved in 1.38g (1.3eq) 4-phenyl-3H-1,2,4-triazole-3,5-diketone ethyl acetate solution under stirring at room temperature, reaction 2h, 40-50℃ reduced pressure concentration to oil, add 100ml ethanol to reflux dissolution, drop to 20℃ crystallization, filtration, further drop to 15℃ crystallization, filtration, drying to get 4.73g yeast sterol, purity 94.8%, as shown in Figure 11 .

[0100] The above only is the preferred embodiment of the present application, should be noted that, for the ordinary skilled in the art, without departing from the principles of the present application, can also make a number of improvements and refinements, these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A process for the preparation of zymosterol, characterized in that, The method comprises the following steps: Step 1: obtaining a crystallization mother liquor after extracting ergosterol from yeast, concentrating, and collecting the concentrate; Step 2: dissolving the concentrate in ethyl acetate, washing with water, separating, and collecting the organic phase; Step 3: reacting the organic phase with 4-phenyl-3H-1, 2, 4-triazole-3, 5-dione, concentrating, dissolving in ethanol, first cooling crystallization, collecting the filtrate by suction filtration, second cooling crystallization, filtering, and drying to obtain the yeast sterol; The crystallization in step 3 comprises first crystallization and second crystallization; the temperature of the first crystallization is 20-25℃; and the temperature of the second crystallization is 0-10℃; The amount of 4-phenyl-3H-1, 2, 4-triazole-3, 5-dione used in step 3 is 1.1-1.5 equivalents; The purity of the 4-phenyl-3H-1, 2, 4-triazole-3, 5-dione is not less than 94%.

2. The production method according to claim 1, wherein The mass ratio of ergosterol in the crystallization mother liquor to 4-phenyl-3H-1, 2, 4-triazole-3, 5-dione is 2.4: (1.16-1.59).

3. The production method according to claim 1, wherein The mass ratio of ergosterol in the crystallization mother liquor to 4-phenyl-3H-1, 2, 4-triazole-3, 5-dione is 2.4:1.16, 2.4:1.38, or 2.4:1.

59.

4. The production method according to claim 1, wherein The first concentration in step 1 is concentration under reduced pressure at 50-60℃.

5. The production method according to claim 1, wherein The temperature of the reaction in step 3 is 25℃±5℃, and the reaction time is 2h.

6. The production method according to claim 1, wherein The concentration in step 3 is concentration under reduced pressure at 40-50℃.

7. The production method according to claim 1, wherein The temperature of the dissolving in step 3 is 70-80℃.

8. The production method according to claim 1, wherein The temperature of the first cooling crystallization in step 3 is 20-25℃.

9. The production method according to claim 1, wherein The temperature of the second cooling crystallization in step 3 is 0-10℃.

10. The production method according to claim 1, wherein The preparation method of the crystallization mother liquor after extracting ergosterol from yeast in step 1 comprises: Step 1-1: subjecting yeast to saponification reaction in alkali and water, performing solid-liquid separation, and collecting a first solid phase; Step 1-2: adding a first solvent to the first solid phase to extract ergosterol, performing solid-liquid separation to obtain a second liquid phase and a second solid phase, and drying the second solid phase to obtain defatted yeast; Step 1-3: concentrating the second liquid phase under reduced pressure to dryness, dissolving in a second solvent, washing with water, separating, collecting the organic layer, concentrating under reduced pressure, adding ethanol to crystallize ergosterol, and collecting the crystallization mother liquor to obtain the crystallization mother liquor after extracting ergosterol from yeast; In step 1-2, the first solvent comprises any one or more of ethanol, n-propanol, isopropanol, n-butanol, acetone, and acetonitrile; In step 1-3, the second solvent comprises any one or more of n-hexane, toluene, ethyl acetate, petroleum ether, cyclohexane, methyl tert-butyl ether, dichloromethane, and chloroform.

11. The production method according to claim 10, wherein In step 1-1, the alkali comprises one or more of potassium hydroxide, sodium hydroxide, lithium hydroxide, sodium ethoxide, and sodium methoxide aqueous solution; In step 1-1, the mass ratio of the yeast, the alkali, and water is 1: (0.1-0.5): (5-15); In step 1-1, the saponification reaction time is 4-12 hours, and the saponification reaction temperature is 60-100℃.

12. The production method according to claim 10, wherein In step 1-1, the temperature of the saponification reaction is 60-80°C.

13. The production method according to claim 10, wherein In step 1-2, the mass-volume ratio of the yeast to the first solvent is 1:(10-20) in kg / L. In step 1-2, the extraction temperature is 56-120°C, and the reflux time is 1-3 hours.

14. The production method according to claim 10, wherein In step 1-2, the mass ratio of the yeast, the alkali substance, and water is 1:(0.2-0.3):(10-13).

15. The production method according to claim 10, wherein In step 1-3, the amount of the second solvent is twice the volume of the yeast. In step 1-3, the temperature of the vacuum concentration is 40-80°C. In step 1-3, the mass ratio of the ethanol to the yeast for crystallization is (0.5-1.5), and the crystallization temperature is 0-30°C.

16. The production method according to claim 1, wherein The yeast includes one or more of Saccharomyces cerevisiae, Candida, Torulopsis, Rhodotorula, or Cryptococcus.

17. A process for the production of ergosterol and zymosterol by coproduction from yeast, characterized in that, The method comprises the following steps: extracting ergosterol from yeast; collecting the crystallization mother liquor after the yeast extracts ergosterol, and obtaining yeast sterol by the preparation method of any one of claims 1-16.

18. The method of claim 17, wherein, The method for extracting ergosterol from yeast comprises: Step 1-1: subjecting yeast to saponification reaction in an alkali substance and water, performing solid-liquid separation, and collecting a first solid phase; Step 1-2: adding a first solvent to the first solid phase to extract ergosterol, performing solid-liquid separation to obtain a second liquid phase and a second solid phase, and drying the second solid phase to obtain defatted yeast; Step 1-3: vacuum concentrating the second liquid phase to dryness, adding a second solvent to dissolve, performing water washing and liquid separation, collecting the organic layer, vacuum concentrating, adding ethanol to crystallize, and obtaining ergosterol; In step 1-2, the first solvent includes any one or more of ethanol, n-propanol, isopropanol, n-butanol, acetone, and acetonitrile. In step 1-3, the second solvent includes any one or more of n-hexane, toluene, ethyl acetate, petroleum ether, cyclohexane, methyl tert-butyl ether, dichloromethane, and chloroform.

19. The method of claim 17, wherein, In step 1-1, the alkali substance includes one or more of potassium hydroxide, sodium hydroxide, lithium hydroxide, sodium ethoxide, and sodium methoxide aqueous solution. In step 1-1, the mass ratio of the yeast, the alkali substance, and water is 1:(0.1-0.5):(5-15). In step 1-1, the saponification reaction time is 4-12 hours, and the saponification reaction temperature is 60-100°C. In step 1-1, the saponification reaction temperature is 60-80°C. In step 1-2, the mass-volume ratio of the yeast to the first solvent is 1:(10-20) in kg / L. In step 1-2, the extraction temperature is 56-120°C, and the reflux time is 1-3 hours. In step 1-2, the mass ratio of the yeast, the alkali substance, and water is 1:(0.2-0.3):(10-13). In step 1-3, the amount of the second solvent is twice the volume of the yeast. In step 1-3, the temperature of the vacuum concentration is 40-80°C. In step 1-3, the mass ratio of the ethanol to the yeast for crystallization is (0.5-1.5), and the crystallization temperature is 0-30°C.

20. The method of claim 17, wherein, The yeast includes one or more of Saccharomyces cerevisiae, Candida, Torulopsis, Rhodotorula, or Cryptococcus. The yeast includes one or more of Saccharomyces cerevisiae, Candida, Torulopsis, Rhodotorula, or Cryptococcus.

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