CN118256575A - Preparation method of composite yeast polysaccharide nucleotide containing multiple organic acids - Google Patents
Preparation method of composite yeast polysaccharide nucleotide containing multiple organic acids Download PDFInfo
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- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 91
- 239000002773 nucleotide Substances 0.000 title claims abstract description 47
- 150000007524 organic acids Chemical class 0.000 title claims abstract description 36
- 235000005985 organic acids Nutrition 0.000 title claims abstract description 36
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 31
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 31
- 239000002131 composite material Substances 0.000 title claims abstract description 29
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims abstract description 54
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims abstract description 54
- 229920000392 Zymosan Polymers 0.000 claims abstract description 48
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 claims abstract description 39
- 229920002498 Beta-glucan Polymers 0.000 claims abstract description 39
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 29
- XJMWHXZUIGHOBA-UHFFFAOYSA-N azane;propanoic acid Chemical compound N.CCC(O)=O XJMWHXZUIGHOBA-UHFFFAOYSA-N 0.000 claims abstract description 24
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 22
- 239000008213 purified water Substances 0.000 claims abstract description 22
- 229910017053 inorganic salt Inorganic materials 0.000 claims abstract description 19
- 235000006408 oxalic acid Nutrition 0.000 claims abstract description 18
- 239000002994 raw material Substances 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 17
- WLJVXDMOQOGPHL-UHFFFAOYSA-N phenylacetic acid Chemical compound OC(=O)CC1=CC=CC=C1 WLJVXDMOQOGPHL-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000000463 material Substances 0.000 claims abstract description 14
- 150000001720 carbohydrates Chemical class 0.000 claims abstract description 12
- WLJVXDMOQOGPHL-PPJXEINESA-N 2-phenylacetic acid Chemical compound O[14C](=O)CC1=CC=CC=C1 WLJVXDMOQOGPHL-PPJXEINESA-N 0.000 claims abstract description 10
- 239000003279 phenylacetic acid Substances 0.000 claims abstract description 8
- 229960003424 phenylacetic acid Drugs 0.000 claims abstract description 8
- 238000000746 purification Methods 0.000 claims abstract description 6
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 88
- 238000002156 mixing Methods 0.000 claims description 44
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 42
- 238000003756 stirring Methods 0.000 claims description 42
- 238000000855 fermentation Methods 0.000 claims description 36
- 230000004151 fermentation Effects 0.000 claims description 36
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 28
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 28
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 28
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 21
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 21
- 238000001816 cooling Methods 0.000 claims description 21
- 238000010438 heat treatment Methods 0.000 claims description 21
- 239000002244 precipitate Substances 0.000 claims description 19
- 230000000694 effects Effects 0.000 claims description 15
- 239000000047 product Substances 0.000 claims description 15
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical group [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 14
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 14
- 239000001110 calcium chloride Substances 0.000 claims description 14
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 14
- 239000011780 sodium chloride Substances 0.000 claims description 14
- 150000001875 compounds Chemical class 0.000 claims description 10
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 7
- 229930006000 Sucrose Natural products 0.000 claims description 7
- 239000008367 deionised water Substances 0.000 claims description 7
- 229910021641 deionized water Inorganic materials 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 7
- 238000004108 freeze drying Methods 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 238000003672 processing method Methods 0.000 claims description 7
- 239000002904 solvent Substances 0.000 claims description 7
- 239000005720 sucrose Substances 0.000 claims description 7
- 238000002137 ultrasound extraction Methods 0.000 claims description 7
- 238000005406 washing Methods 0.000 claims description 7
- 150000004676 glycans Chemical class 0.000 claims description 6
- 238000000926 separation method Methods 0.000 claims description 4
- 229920002261 Corn starch Polymers 0.000 claims description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 2
- 239000008120 corn starch Substances 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 238000001556 precipitation Methods 0.000 claims description 2
- 238000001179 sorption measurement Methods 0.000 claims description 2
- 239000012535 impurity Substances 0.000 abstract description 6
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 238000003889 chemical engineering Methods 0.000 abstract description 2
- 239000012847 fine chemical Substances 0.000 abstract description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 9
- 150000003839 salts Chemical class 0.000 description 5
- 125000000185 sucrose group Chemical group 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000002777 nucleoside Substances 0.000 description 3
- 150000003833 nucleoside derivatives Chemical class 0.000 description 3
- -1 phosphite triester Chemical class 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000012454 non-polar solvent Substances 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000002798 polar solvent Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
Abstract
The invention belongs to the technical field of fine chemical engineering, in particular to a preparation method of composite yeast polysaccharide nucleotide containing various organic acids, which aims at the problems of low production efficiency, low purity and large amount of impurities mixed in the existing method, and provides the following scheme that the method comprises the following steps: s1: the material consists of the following raw materials in parts by weight: 15-20 parts of citric acid, 10-15 parts of oxalic acid, 5-10 parts of ammonium propionate, 2-7 parts of phenylacetic acid, 25-30 parts of zymosan, 10-15 parts of yeast beta-glucan extract, 10-15 parts of saccharides and 5-10 parts of inorganic salt; s2: the citric acid, oxalic acid and phenylacetic acid are dissolved in purified water at 25-35 ℃ to obtain a solution, more composite zymosan nucleotides containing various organic acids can be produced by the same amount of materials, and the impurity content in the composite zymosan nucleotides containing various organic acids is reduced after purification treatment.
Description
Technical Field
The invention relates to the technical field of fine chemical engineering, in particular to a preparation method of composite yeast polysaccharide nucleotide containing various organic acids.
Background
The Chinese patent document with the application number 201680074787.3 discloses a preparation method of an oligonucleotide and a nucleoside, a nucleotide or an oligonucleotide, which comprises the following steps: (1) A step of condensing a nucleoside, nucleotide or oligonucleotide (a) or the like in which the 5 '-hydroxyl group is not protected with a nucleoside, nucleotide or oligonucleotide (b) in which the 5' -hydroxyl group is protected in a nonpolar solvent to obtain a reaction solution containing a phosphite triester product (c); (3) A step of oxidizing or sulfiding the phosphite triester product (c) to obtain a reaction solution containing the oligonucleotide (d) in which the 5' -hydroxyl group is protected; (4) A step of deprotecting the oligonucleotide (d) to obtain a reaction solution containing the oligonucleotide (e) in which the 5' -hydroxyl group is not protected; and (6) a step of adding a polar solvent to the reaction liquid containing the oligonucleotide (e) and purifying the oligonucleotide (e) by solid-liquid separation or extraction.
However, the above technical solution also has some problems, for example, the amount of the produced compound yeast polysaccharide nucleotide is small, the proportion of the raw material converted into the compound yeast polysaccharide nucleotide is low, which affects the production efficiency of the compound yeast polysaccharide nucleotide, and the purity of the compound yeast polysaccharide nucleotide is low, so that a large amount of impurities are mixed.
Disclosure of Invention
Based on the problems of low production efficiency, low purity and large amount of impurities in the prior art, the invention provides a preparation method of a composite yeast polysaccharide nucleotide containing various organic acids.
The invention provides a preparation method of a composite yeast polysaccharide nucleotide containing various organic acids, which comprises the following steps:
S1: the material consists of the following raw materials in parts by weight: 15-20 parts of citric acid, 10-15 parts of oxalic acid, 5-10 parts of ammonium propionate, 2-7 parts of phenylacetic acid, 25-30 parts of zymosan, 10-15 parts of yeast beta-glucan extract, 10-15 parts of saccharides and 5-10 parts of inorganic salt;
S2: dissolving citric acid, oxalic acid and phenylacetic acid in purified water at 25-35 ℃ to obtain a solution, adding ammonium propionate under the negative pressure condition, and uniformly mixing to obtain a mixture;
s3: adding zymosan and yeast beta-glucan extract at 20-35 ℃, uniformly mixing, adding sugar and inorganic salt, continuously mixing, and mixing to obtain a fermentation product;
S4: the ferment is poured into a fermenter for fermentation, and after the fermentation is completed, the composite zymosan nucleotide containing various organic acids is obtained by separation and purification, the composite zymosan nucleotide containing various organic acids is produced by raw materials such as citric acid, oxalic acid, ammonium propionate, phenylacetic acid, zymosan, yeast beta-glucan extract, saccharides, inorganic salts and the like, more composite zymosan nucleotide containing various organic acids can be produced by the same amount of materials, and the impurity content in the composite zymosan nucleotide containing various organic acids is reduced by purification treatment.
Preferably, in S1, the raw materials of the yeast β -glucan extract include highly active baker' S yeast, sodium chloride, hydrochloric acid, purified water and ethyl acetate, so that the yeast β -glucan extract can be effectively extracted.
Preferably, the preparation method of the yeast beta-glucan extract comprises the following steps: adding sodium chloride, purified water and ethyl acetate into the high-activity baker's yeast, stirring and dispersing, adding hydrochloric acid into the baker's yeast to adjust the pH, heating to 50-55 ℃, preserving heat and mixing for 25 hours, stopping heating, naturally cooling, cooling to room temperature, cooling to 3 ℃ by using a cold source, centrifuging for 10 minutes, taking precipitate, washing with deionized water for 4 times, drying, finally dissolving the precipitate in sodium hydroxide solution, and performing ultrasonic extraction to obtain yeast beta-glucan extract, thereby ensuring the purity.
Preferably, the sugar is one of corn starch and sucrose, and the sugar is crushed to be 200 meshes by a crushing grinder, so that saccharification of the material is facilitated.
Preferably, in the step S1, the inorganic salt is calcium chloride and calcium carbonate, and the weight ratio of the calcium chloride to the calcium carbonate is 2:1, so that the production of the material is facilitated.
Preferably, in S1, the processing method of zymosan includes: pouring the zymosan into a potassium hydroxide solvent for mixing and dispersing, heating to 65-75 ℃ after wall breaking treatment, adjusting the pH value to 7, purifying by using 30wt% hydrogen peroxide solution, and freeze-drying to obtain purified zymosan, thereby ensuring the effect of the zymosan.
Preferably, in the step S2, the stirring speed is controlled to be 240 r/min-360 r/min after the ammonium propionate is added, the stirring temperature is controlled to be 35-45 ℃, and the stirring is carried out for 15 minutes, so that the mixing efficiency is ensured.
Preferably, in the step S3, the stirring speed is controlled to be 450-600 r/min, the stirring temperature is controlled to be 25-40 ℃, and the mixing effect is ensured.
Preferably, in the step S4, the fermentation temperature is controlled to be 25-35 ℃, and the fermentation time is controlled to be 12-24 hours, so that materials can be fully fused.
Preferably, in S4, one or two of a centrifuge, a filtering device, a precipitation device and an adsorption device are used for separation and purification, so as to ensure the purification effect.
The invention has the beneficial effects that:
The composite zymosan nucleotide containing various organic acids is produced by using citric acid, oxalic acid, ammonium propionate, phenylacetic acid, zymosan, yeast beta-glucan extract, saccharides, inorganic salts and other raw materials, more composite zymosan nucleotide containing various organic acids can be produced by using the same amount of materials, and the impurity content in the composite zymosan nucleotide containing various organic acids is reduced by purifying treatment.
Drawings
Fig. 1 is a flow chart of the operation proposed by the present invention.
Detailed Description
The invention is further illustrated below in connection with specific embodiments.
Referring to fig. 1, embodiment one
In this embodiment, a method for preparing a composite yeast polysaccharide nucleotide containing multiple organic acids is provided, which comprises the following steps:
s1: the material consists of the following raw materials in parts by weight: 16 parts of citric acid, 14 parts of oxalic acid, 7 parts of ammonium propionate, 3 parts of phenylacetic acid, 30 parts of zymosan, 10 parts of yeast beta-glucan extract, 12 parts of saccharides and 8 parts of inorganic salts, wherein the raw materials of the yeast beta-glucan extract comprise high-activity baker's yeast, sodium chloride, hydrochloric acid, purified water and ethyl acetate, and the preparation method of the yeast beta-glucan extract comprises the following steps: adding sodium chloride, purified water and ethyl acetate into high-activity baker's yeast, stirring and dispersing, adding hydrochloric acid into the baker's yeast to adjust the pH, heating to 53 ℃, preserving heat and mixing for 25 hours, stopping heating, naturally cooling, cooling to room temperature, cooling to 3 ℃ by using a cold source, centrifuging for 10 minutes, taking precipitate, washing the precipitate with deionized water for 4 times, drying, finally dissolving the precipitate in sodium hydroxide solution, performing ultrasonic extraction to obtain yeast beta-glucan extract, wherein sugar is sucrose, the sugar is crushed to 200 meshes by using a crushing grinder, inorganic salt is calcium chloride and calcium carbonate, the weight ratio of the calcium chloride to the calcium carbonate is 2:1, and the processing method of yeast polysaccharide comprises the steps of: pouring the zymosan into a potassium hydroxide solvent for mixing and dispersing, heating to 69 ℃ after wall breaking treatment, adjusting the pH value to 7, purifying by using 30wt% hydrogen peroxide solution, and freeze-drying to obtain purified zymosan;
S2: dissolving citric acid, oxalic acid and phenylacetic acid in purified water at 25-35 ℃ to obtain a solution, adding ammonium propionate under negative pressure, uniformly mixing to obtain a mixture, adding ammonium propionate, controlling the stirring speed at 320r/min, controlling the stirring temperature at 42 ℃, and stirring for 15 minutes;
S3: adding zymosan and yeast beta-glucan extract at 20-35 ℃, uniformly mixing, adding sugar and inorganic salt, continuously mixing, and obtaining a fermented product by mixing, wherein the stirring speed is controlled at 560r/min, and the stirring temperature is controlled at 31 ℃;
S4: pouring the fermented product into a fermentation tank for fermentation, separating and purifying after the fermentation is completed to obtain the compound zymosan nucleotide containing various organic acids, controlling the fermentation temperature at 31 ℃ and the fermentation time at 17 hours, and separating and purifying by adopting a centrifuge.
Referring to FIG. 1, embodiment II
In this embodiment, a method for preparing a composite yeast polysaccharide nucleotide containing multiple organic acids is provided, which comprises the following steps:
S1: the material consists of the following raw materials in parts by weight: 19 parts of citric acid, 11 parts of oxalic acid, 6 parts of ammonium propionate, 4 parts of phenylacetic acid, 26 parts of zymosan, 14 parts of yeast beta-glucan extract, 11 parts of saccharides and 9 parts of inorganic salts, wherein the raw materials of the yeast beta-glucan extract comprise high-activity baker's yeast, sodium chloride, hydrochloric acid, purified water and ethyl acetate, and the preparation method of the yeast beta-glucan extract comprises the following steps: adding sodium chloride, purified water and ethyl acetate into high-activity baker's yeast, stirring and dispersing, adding hydrochloric acid into the baker's yeast to adjust the pH, heating to 53 ℃, preserving heat and mixing for 25 hours, stopping heating, naturally cooling, cooling to room temperature, cooling to 3 ℃ by using a cold source, centrifuging for 10 minutes, taking precipitate, washing the precipitate with deionized water for 4 times, drying, finally dissolving the precipitate in sodium hydroxide solution, performing ultrasonic extraction to obtain yeast beta-glucan extract, wherein sugar is sucrose, the sugar is crushed to 200 meshes by using a crushing grinder, inorganic salt is calcium chloride and calcium carbonate, the weight ratio of the calcium chloride to the calcium carbonate is 2:1, and the processing method of yeast polysaccharide comprises the steps of: pouring the zymosan into a potassium hydroxide solvent for mixing and dispersing, heating to 69 ℃ after wall breaking treatment, adjusting the pH value to 7, purifying by using 30wt% hydrogen peroxide solution, and freeze-drying to obtain purified zymosan;
S2: dissolving citric acid, oxalic acid and phenylacetic acid in purified water at 25-35 ℃ to obtain a solution, adding ammonium propionate under negative pressure, uniformly mixing to obtain a mixture, adding ammonium propionate, controlling the stirring speed at 320r/min, controlling the stirring temperature at 42 ℃, and stirring for 15 minutes;
S3: adding zymosan and yeast beta-glucan extract at 20-35 ℃, uniformly mixing, adding sugar and inorganic salt, continuously mixing, and obtaining a fermented product by mixing, wherein the stirring speed is controlled at 560r/min, and the stirring temperature is controlled at 31 ℃;
S4: pouring the fermented product into a fermentation tank for fermentation, separating and purifying after the fermentation is completed to obtain the compound zymosan nucleotide containing various organic acids, controlling the fermentation temperature at 31 ℃ and the fermentation time at 17 hours, and separating and purifying by adopting a centrifuge.
Referring to fig. 1, embodiment three
In this embodiment, a method for preparing a composite yeast polysaccharide nucleotide containing multiple organic acids is provided, which comprises the following steps:
S1: the material consists of the following raw materials in parts by weight: 18 parts of citric acid, 12 parts of oxalic acid, 6 parts of ammonium propionate, 4 parts of phenylacetic acid, 28 parts of zymosan, 12 parts of yeast beta-glucan extract, 13 parts of saccharides and 7 parts of inorganic salt, wherein the raw materials of the yeast beta-glucan extract comprise high-activity baker's yeast, sodium chloride, hydrochloric acid, purified water and ethyl acetate, and the preparation method of the yeast beta-glucan extract comprises the following steps: adding sodium chloride, purified water and ethyl acetate into high-activity baker's yeast, stirring and dispersing, adding hydrochloric acid into the baker's yeast to adjust the pH, heating to 53 ℃, preserving heat and mixing for 25 hours, stopping heating, naturally cooling, cooling to room temperature, cooling to 3 ℃ by using a cold source, centrifuging for 10 minutes, taking precipitate, washing the precipitate with deionized water for 4 times, drying, finally dissolving the precipitate in sodium hydroxide solution, performing ultrasonic extraction to obtain yeast beta-glucan extract, wherein sugar is sucrose, the sugar is crushed to 200 meshes by using a crushing grinder, inorganic salt is calcium chloride and calcium carbonate, the weight ratio of the calcium chloride to the calcium carbonate is 2:1, and the processing method of yeast polysaccharide comprises the steps of: pouring the zymosan into a potassium hydroxide solvent for mixing and dispersing, heating to 69 ℃ after wall breaking treatment, adjusting the pH value to 7, purifying by using 30wt% hydrogen peroxide solution, and freeze-drying to obtain purified zymosan;
S2: dissolving citric acid, oxalic acid and phenylacetic acid in purified water at 25-35 ℃ to obtain a solution, adding ammonium propionate under negative pressure, uniformly mixing to obtain a mixture, adding ammonium propionate, controlling the stirring speed at 320r/min, controlling the stirring temperature at 42 ℃, and stirring for 15 minutes;
S3: adding zymosan and yeast beta-glucan extract at 20-35 ℃, uniformly mixing, adding sugar and inorganic salt, continuously mixing, and obtaining a fermented product by mixing, wherein the stirring speed is controlled at 560r/min, and the stirring temperature is controlled at 31 ℃;
S4: pouring the fermented product into a fermentation tank for fermentation, separating and purifying after the fermentation is completed to obtain the compound zymosan nucleotide containing various organic acids, controlling the fermentation temperature at 31 ℃ and the fermentation time at 17 hours, and separating and purifying by adopting a centrifuge.
Referring to fig. 1, embodiment four
In this embodiment, a method for preparing a composite yeast polysaccharide nucleotide containing multiple organic acids is provided, which comprises the following steps:
S1: the material consists of the following raw materials in parts by weight: 15 parts of citric acid, 14 parts of oxalic acid, 6 parts of ammonium propionate, 7 parts of phenylacetic acid, 28 parts of zymosan, 12 parts of yeast beta-glucan extract, 12 parts of saccharides and 6 parts of inorganic salts, wherein the raw materials of the yeast beta-glucan extract comprise high-activity baker's yeast, sodium chloride, hydrochloric acid, purified water and ethyl acetate, and the preparation method of the yeast beta-glucan extract comprises the following steps: adding sodium chloride, purified water and ethyl acetate into high-activity baker's yeast, stirring and dispersing, adding hydrochloric acid into the baker's yeast to adjust the pH, heating to 53 ℃, preserving heat and mixing for 25 hours, stopping heating, naturally cooling, cooling to room temperature, cooling to 3 ℃ by using a cold source, centrifuging for 10 minutes, taking precipitate, washing the precipitate with deionized water for 4 times, drying, finally dissolving the precipitate in sodium hydroxide solution, performing ultrasonic extraction to obtain yeast beta-glucan extract, wherein sugar is sucrose, the sugar is crushed to 200 meshes by using a crushing grinder, inorganic salt is calcium chloride and calcium carbonate, the weight ratio of the calcium chloride to the calcium carbonate is 2:1, and the processing method of yeast polysaccharide comprises the steps of: pouring the zymosan into a potassium hydroxide solvent for mixing and dispersing, heating to 69 ℃ after wall breaking treatment, adjusting the pH value to 7, purifying by using 30wt% hydrogen peroxide solution, and freeze-drying to obtain purified zymosan;
S2: dissolving citric acid, oxalic acid and phenylacetic acid in purified water at 25-35 ℃ to obtain a solution, adding ammonium propionate under negative pressure, uniformly mixing to obtain a mixture, adding ammonium propionate, controlling the stirring speed at 320r/min, controlling the stirring temperature at 42 ℃, and stirring for 15 minutes;
S3: adding zymosan and yeast beta-glucan extract at 20-35 ℃, uniformly mixing, adding sugar and inorganic salt, continuously mixing, and obtaining a fermented product by mixing, wherein the stirring speed is controlled at 560r/min, and the stirring temperature is controlled at 31 ℃;
S4: pouring the fermented product into a fermentation tank for fermentation, separating and purifying after the fermentation is completed to obtain the compound zymosan nucleotide containing various organic acids, controlling the fermentation temperature at 31 ℃ and the fermentation time at 17 hours, and separating and purifying by adopting a centrifuge.
Referring to fig. 1, embodiment five
In this embodiment, a method for preparing a composite yeast polysaccharide nucleotide containing multiple organic acids is provided, which comprises the following steps:
S1: the material consists of the following raw materials in parts by weight: 15 parts of citric acid, 15 parts of oxalic acid, 8 parts of ammonium propionate, 2 parts of phenylacetic acid, 30 parts of zymosan, 15 parts of yeast beta-glucan extract, 10 parts of saccharides and 5 parts of inorganic salt, wherein the raw materials of the yeast beta-glucan extract comprise high-activity baker's yeast, sodium chloride, hydrochloric acid, purified water and ethyl acetate, and the preparation method of the yeast beta-glucan extract comprises the following steps: adding sodium chloride, purified water and ethyl acetate into high-activity baker's yeast, stirring and dispersing, adding hydrochloric acid into the baker's yeast to adjust the pH, heating to 53 ℃, preserving heat and mixing for 25 hours, stopping heating, naturally cooling, cooling to room temperature, cooling to 3 ℃ by using a cold source, centrifuging for 10 minutes, taking precipitate, washing the precipitate with deionized water for 4 times, drying, finally dissolving the precipitate in sodium hydroxide solution, performing ultrasonic extraction to obtain yeast beta-glucan extract, wherein sugar is sucrose, the sugar is crushed to 200 meshes by using a crushing grinder, inorganic salt is calcium chloride and calcium carbonate, the weight ratio of the calcium chloride to the calcium carbonate is 2:1, and the processing method of yeast polysaccharide comprises the steps of: pouring the zymosan into a potassium hydroxide solvent for mixing and dispersing, heating to 69 ℃ after wall breaking treatment, adjusting the pH value to 7, purifying by using 30wt% hydrogen peroxide solution, and freeze-drying to obtain purified zymosan;
S2: dissolving citric acid, oxalic acid and phenylacetic acid in purified water at 25-35 ℃ to obtain a solution, adding ammonium propionate under negative pressure, uniformly mixing to obtain a mixture, adding ammonium propionate, controlling the stirring speed at 320r/min, controlling the stirring temperature at 42 ℃, and stirring for 15 minutes;
S3: adding zymosan and yeast beta-glucan extract at 20-35 ℃, uniformly mixing, adding sugar and inorganic salt, continuously mixing, and obtaining a fermented product by mixing, wherein the stirring speed is controlled at 560r/min, and the stirring temperature is controlled at 31 ℃;
S4: pouring the fermented product into a fermentation tank for fermentation, separating and purifying after the fermentation is completed to obtain the compound zymosan nucleotide containing various organic acids, controlling the fermentation temperature at 31 ℃ and the fermentation time at 17 hours, and separating and purifying by adopting a centrifuge.
Comparing conventional nucleotides with the nucleotides prepared in examples one to five, the nucleotides prepared in examples one to five are as follows:
The comparative example selects the patent with application number 201680074787.3.
As can be seen from the above table, the yield and purity of the nucleotide prepared by the present invention are significantly improved, and the second preferred embodiment is implemented.
The foregoing is only a preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art, who is within the scope of the present invention, should make equivalent substitutions or modifications according to the technical scheme of the present invention and the inventive concept thereof, and should be covered by the scope of the present invention.
Claims (10)
1. The preparation method of the composite yeast polysaccharide nucleotide containing a plurality of organic acids is characterized by comprising the following steps:
S1: the material consists of the following raw materials in parts by weight: 15-20 parts of citric acid, 10-15 parts of oxalic acid, 5-10 parts of ammonium propionate, 2-7 parts of phenylacetic acid, 25-30 parts of zymosan, 10-15 parts of yeast beta-glucan extract, 10-15 parts of saccharides and 5-10 parts of inorganic salt;
S2: dissolving citric acid, oxalic acid and phenylacetic acid in purified water at 25-35 ℃ to obtain a solution, adding ammonium propionate under the negative pressure condition, and uniformly mixing to obtain a mixture;
s3: adding zymosan and yeast beta-glucan extract at 20-35 ℃, uniformly mixing, adding sugar and inorganic salt, continuously mixing, and mixing to obtain a fermentation product;
S4: pouring the fermented product into a fermentation tank for fermentation, and separating and purifying after the fermentation is completed to obtain the compound zymosan nucleotide containing various organic acids.
2. The method for preparing a composite yeast polysaccharide nucleotide containing a plurality of organic acids according to claim 1, wherein in S1, the raw materials of the yeast beta-glucan extract comprise highly active baker' S yeast, sodium chloride, hydrochloric acid, purified water and ethyl acetate.
3. The method for preparing a composite yeast polysaccharide nucleotide containing a plurality of organic acids according to claim 2, wherein the method for preparing yeast beta-glucan extract is as follows: adding sodium chloride, purified water and ethyl acetate into the high-activity baker's yeast, stirring and dispersing, adding hydrochloric acid into the baker's yeast to adjust the pH, heating to 50-55 ℃, preserving heat and mixing for 25 hours, stopping heating, naturally cooling, cooling to room temperature, cooling to 3 ℃ by using a cold source, centrifuging for 10 minutes, taking precipitate, washing with deionized water for 4 times, drying, finally dissolving the precipitate in sodium hydroxide solution, and carrying out ultrasonic extraction to obtain the yeast beta-glucan extract.
4. The method for preparing a composite yeast polysaccharide nucleotide containing a plurality of organic acids according to claim 1, wherein in S1, the saccharide is one of corn starch and sucrose, and the saccharide is crushed to 200 mesh by a crushing grinder.
5. The method for preparing a composite yeast polysaccharide nucleotide containing a plurality of organic acids according to claim 1, wherein in S1, the inorganic salt is calcium chloride and calcium carbonate, and the weight ratio of the calcium chloride to the calcium carbonate is 2:1.
6. The method for preparing a composite yeast polysaccharide nucleotide containing a plurality of organic acids according to claim 1, wherein in S1, the processing method of the yeast polysaccharide comprises: pouring the zymosan into a potassium hydroxide solvent for mixing and dispersing, heating to 65-75 ℃ after wall breaking treatment, adjusting the pH value to 7, purifying by using 30wt% hydrogen peroxide solution, and freeze-drying to obtain the purified zymosan.
7. The method for preparing a composite yeast polysaccharide nucleotide containing a plurality of organic acids according to claim 1, wherein in the step S2, the stirring speed is controlled to be 240 r/min-360 r/min after adding ammonium propionate, the stirring temperature is controlled to be 35-45 ℃, and the stirring time is 15 minutes.
8. The method for preparing a composite yeast polysaccharide nucleotide containing a plurality of organic acids according to claim 1, wherein in the step S3, the stirring speed is controlled to be 450-600 r/min, and the stirring temperature is controlled to be 25-40 ℃.
9. The method for preparing a composite yeast polysaccharide nucleotide containing a plurality of organic acids according to claim 1, wherein in the step S4, the fermentation temperature is controlled to be 25-35 ℃, and the fermentation time is controlled to be 12-24 hours.
10. The method for preparing a complex yeast polysaccharide nucleotide containing a plurality of organic acids according to claim 1, wherein in S4, separation and purification are performed by using one or two of a centrifuge, a filtering device, a precipitation device and an adsorption device.
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