CN110818753A - Method for recycling crystallized trehalose mother liquor - Google Patents
Method for recycling crystallized trehalose mother liquor Download PDFInfo
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- CN110818753A CN110818753A CN201911105271.2A CN201911105271A CN110818753A CN 110818753 A CN110818753 A CN 110818753A CN 201911105271 A CN201911105271 A CN 201911105271A CN 110818753 A CN110818753 A CN 110818753A
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- trehalose
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/04—Disaccharides
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/02—Monosaccharides
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/12—Disaccharides
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
Abstract
The invention discloses a method for recycling crystallized trehalose mother liquor, and belongs to the technical field of food. The invention saccharifies trisaccharide and oligosaccharide above trisaccharide in the mother liquor after chromatographic separation into glucose by using an enzyme preparation, hydrolyzes soluble protein by using protease, removes impurities by decoloring and ion exchange, and the crystallized trehalose finished product reaches the national standard requirement, and the purity can reach 99.5 percent to the maximum. The efficient and high-added-value utilization of the crystallization mother liquor in the trehalose production process is realized.
Description
Technical Field
The invention relates to a method for recycling crystallized trehalose mother liquor, and belongs to the technical field of food.
Background
Trehalose is a non-reducing disaccharide formed by connecting two pyran ring glucose molecules by α -1,1 glucoside cans, can stabilize the structures of cells and proteins under severe environments of high temperature, high cold, high osmotic pressure, drying dehydration and the like, and effectively protects the cells and the protein molecules from being inactivated without degeneration, thereby maintaining the life process and biological characteristics of organisms, and the trehalose is widely applied to the fields of food, agriculture, medicine and the like.
At present, trehalose is produced at home and abroad by adopting an enzyme method, the trehalose is separated after cooling and crystallization, a large amount of mother liquor is remained, and a common mother liquor treatment method comprises 1) directly using the trehalose as the mother liquor to buy at a low price, and 2) carrying out recrystallization after separating glucose in the mother liquor by chromatography. The chromatographic separation mother liquor can remove glucose in the mother liquor, at the same time retain polysaccharide and soluble protein over disaccharide, and the components over trisaccharide and trisaccharide can be enriched with the increase of circulation frequency. The sea purity of the mother liquor after chromatographic separation cannot meet the national standard requirement due to the increase of the contents of trisaccharide, oligosaccharide above trisaccharide and soluble protein, and the mother liquor can only be sold and treated at low cost. To address this problem. The mother liquor recycling process needs to be researched, a high-efficiency and high-added-value treatment method is provided for mother liquor treatment after trehalose crystallization and centrifugation, the production cost is reduced, and the product yield is improved.
Disclosure of Invention
The invention aims to provide a method for recycling crystallized trehalose mother liquor, aiming at the defects of the existing mother liquor treatment mode in the trehalose production process, the mother liquor with high content of trisaccharide and components above trisaccharide is saccharified and proteolyzed to obtain micromolecule polypeptide and amino acid, polypeptide and amino acid are removed through decolorization and ion exchange, and then the crystallized trehalose finished product with the purity of more than 99.5 percent is obtained through concentration and crystallization and re-separation.
The first purpose of the invention is to provide a method for recycling crystallized trehalose mother liquor, which comprises the steps of saccharifying and proteolysis treatment of the crystallized trehalose mother liquor, decoloring, ion exchange treatment, concentration and crystallization to obtain the crystallized trehalose.
In one embodiment, the saccharification process is an enzymatic reaction with a saccharifying enzyme and pullulanase; the protease is keratinase.
In one embodiment, the saccharification is carried out by adjusting the trehalose mother liquor to a solution with a solid content of 25-30 and adding an enzyme.
In one embodiment, the saccharification system has a pH of 4.2-4.4 and a saccharification temperature of 58-62 ℃.
In one embodiment, the saccharifying enzyme is added in an amount of 2.4U/kgDS to 10U/kgDS, pullulanase is added in an amount of 0.1U/kgDS to 0.2U/kgDS, and acid keratinase is added in an amount of 10 to 20U/kgDS.
In one embodiment, the enzymolysis condition of the protease is 30-55 ℃, and the enzymolysis time is 6-16 h.
In one embodiment, the ion exchange is performed by using cation exchange resin (D001) and anion exchange resin (D301).
In one embodiment, the crystallization is performed in a temperature reduction process, wherein the temperature reduction is from 50 ℃ to 20 ℃, and the crystallization is performed for 72 hours.
In one embodiment, the method comprises the steps of:
(1) saccharification: centrifuging trehalose, preparing a mother solution into a solution with a solid content of 25-30, heating to 58-62 ℃, adjusting the pH value to 4.2-4.4, adding saccharifying enzyme according to a ratio of 2.4U/kg DS-10U/kg DS (DS is dry matter), adding pullulanase according to a ratio of 0.1U/kg DS-0.2U/kg DS, and saccharifying for 32-72 h;
(2) and (3) proteolysis: cooling to 30-55 deg.C, adding acid keratinase at a ratio of 10-20U/kgDS, and reacting for 6-16h under low-speed stirring;
(3) and (3) decoloring and filtering: heating to inactivate enzyme, adding active carbon into saccharified liquid for decolorization to make sugar liquid clear and transparent, and free of foreign matters and visible impurities;
(4) ion exchange concentration: purifying with cation exchange resin (D001) and anion exchange resin (D301), and concentrating sugar solution to 71-73% by vacuum concentrator;
(5) stirring and crystallizing: adding crystal trehalose seed crystal with the mass of 0.5-1.5%, gradually reducing the temperature from 50 ℃ to 20 ℃, and taking 72 hours;
(6) centrifugal drying: and (4) drying the product for 6 hours in a drying oven at 90 ℃ after centrifugation to obtain the finished product trehalose with the trehalose content of more than 98%.
In one embodiment of the invention, the decolorization filtration is carried out by using activated carbon, and the filtration is carried out by using a plate-and-frame filter press until the sugar solution has no visible impurities and the light transmittance is more than 95 percent.
The second purpose of the invention is to provide the trehalose prepared by the method.
The invention also claims the application of the method in the production of trehalose and derivatives thereof in the fields of food, medicine and chemical industry.
Has the advantages that: the invention saccharifies trisaccharide and oligosaccharide above trisaccharide in the mother liquor after chromatographic separation into glucose by using an enzyme preparation, hydrolyzes soluble protein by using protease, removes impurities by decoloring and ion exchange, and the crystallized trehalose finished product reaches the national standard requirement, and the purity can reach 99.5 percent to the maximum. The efficient and high-added-value utilization of the crystallization mother liquor in the trehalose production process is realized.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more clear, the following will describe embodiments of the present invention in further detail:
pullulanase: 1500U/g, available from Shandong-Long-Kete enzyme preparations, Inc.; saccharifying enzyme was purchased from Novoxin, 100000U/g; the acid keratinase and the acid protease are purchased from Xiasan industry group; flavourzyme and papain were purchased from novacin.
Example 1:
the embodiment provides a recycling process of crystallized trehalose mother liquor, wherein the method comprises the following steps:
1. at 3m3Adding 3m into saccharifying tank3Crystallizing mother liquor (with concentration of BX being 30), heating to 61 ℃, and adjusting pH to 4.3;
2. stirring at low speed, adding saccharifying enzyme at 5.0U/kgDS, adding pullulanase at 0.1U/kgDS, and saccharifying for 72 h;
3. cooling to 55 ℃, adjusting the pH value to 5.5, adding acid keratinase with the enzyme activity of 150U/kgDS into the system reacted in the step (2), and reacting for 12 hours under the condition of low-speed stirring at 80-100 rpm.
4. Heating to inactivate enzyme, adding 0.5% medicinal active carbon by mass into saccharified liquid, decolorizing at 45 deg.C for 30min, and filtering with plate-and-frame filter until the sugar liquid is clear and transparent, has no foreign matter and no visible impurities, and has light transmittance of more than 95%;
5. ion exchange: purifying with cation exchange resin (D001) and anion exchange resin (D301), and concentrating sugar solution with vacuum concentrator until BX is 72;
6. and (3) crystallization: adding crystal trehalose seed crystals accounting for 0.5-1.5% of the mass, cooling to 20 ℃ from 60 ℃, and crystallizing for 72 hours;
7. and (4) drying the product for 6 hours in a drying oven at 90 ℃ after centrifugation to obtain the finished product trehalose with the trehalose content of more than 98%.
Sampling and measuring physical and chemical indexes before and after crystallization:
table 1: mother liquor data before crystallization
| Item | Glucose | Trehalose | Trisaccharides |
| Liquid phase data | 7.819％ | 68.777％ | 16.506％ |
Table 2: data of the final product after crystallization:
| item | GB/T23529-2009 | Detecting data |
| Trehalose content (on a dry basis)/%) | ≥98 | 98.9 |
| pH | 5.0-6.7 | 5.6 |
| Combustion residue/%) | 0.02 | 0.005 |
| Loss on drying is less than or equal to | 1.5 | 0.010 |
| Chroma is less than or equal to | 0.100 | 0.007 |
| Turbidity is less than or equal to | 0.05 | 0.001 |
Example 2:
the embodiment provides a recycling process of crystallized trehalose mother liquor, wherein the method comprises the following steps:
1. at 3m3Adding 3m into saccharifying tank3Crystallizing mother liquor (with concentration of BX being 30), heating to 61 ℃, and adjusting pH to 4.3;
2. stirring at low speed, adding saccharifying enzyme at 5.0U/kgDS, adding pullulanase at 0.2U/kgDS, and saccharifying for 40 h;
3. cooling to 55 ℃, adjusting the pH value to 5.0, adding 0.2g of solid keratinase with the enzyme activity of 200U/kgDS into the system reacted in the step (2), and reacting for 8 hours under the condition of low-speed stirring at 80-100 rpm.
4. Heating to inactivate enzyme, adding 0.5% medicinal active carbon by mass into saccharified liquid, decolorizing at 45 deg.C for 30min, and filtering with plate-and-frame filter until the sugar liquid is clear and transparent, has no foreign matter and no visible impurities, and has light transmittance of more than 95%;
5. ion exchange: purifying with cation exchange resin (D001) and anion exchange resin (D301), concentrating sugar solution to BX 72 with vacuum concentrator, cooling from 60 deg.C to 20 deg.C, and crystallizing for 72 h;
6. and (4) drying the product for 6 hours in a drying oven at 90 ℃ after centrifugation to obtain the finished product trehalose with the trehalose content of more than 99%.
Sampling and measuring physical and chemical indexes before and after crystallization:
table 3: data on detection of crystallized finished product
| Item | GB/T23529-2009 | Detecting data |
| Trehalose content (on a dry basis)/%) | ≥98 | 99.5 |
| pH | 5.0-6.7 | 5.4 |
| Combustion residue/%) | 0.02 | 0.003 |
| Loss on drying is less than or equal to | 1.5 | 0.3 |
| Chroma is less than or equal to | 0.100 | 0.005 |
| Turbidity is less than or equal to | 0.05 | 0.001 |
Comparative example 1:
the embodiment is different from embodiment 1 in that step (3) is omitted.
Comparative example 2:
the specific embodiment is the same as example 1, except that papain is substituted for the acid keratinase.
Comparative example 3:
the specific embodiment is the same as example 1, except that the acid keratinase is replaced with the flavourzyme.
Comparative example 4:
the specific embodiment is the same as example 1 except that the amount of the saccharifying enzyme added in example 1 is adjusted to 2U/kgDS.
Comparative example 5:
the specific embodiment is the same as example 1 except that the reaction time of step (3) is shortened to 6 hours.
Comparative example 6:
the specific embodiment is the same as example 1 except that the cation exchange resin was replaced with (001 × 7) and the anion exchange resin was replaced with D354.
Trehalose prepared by the methods described in comparative examples 1 to 6 was examined and the results are shown in Table 4. The enzymolysis treatment process of the mother liquor plays an important role in the treatment of the trehalose mother liquor, and the full action of protease and saccharifying enzyme can effectively improve the purity of trehalose.
Table 4: detection data of trehalose in finished products obtained under different conditions
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. A method for recycling crystallized trehalose mother liquor is characterized in that the crystallized trehalose mother liquor is subjected to saccharification and proteolysis treatment, decolorization, ion exchange treatment, concentration and crystallization to obtain the crystallized trehalose.
2. The method of claim 1, wherein the saccharification process is a enzymatic hydrolysis reaction with a saccharifying enzyme and a pullulanase; the protease is keratinase.
3. The method according to claim 1, wherein the saccharification is carried out by adjusting the trehalose mother liquor to a solution having a solid content of 25 to 30 and adding an enzyme.
4. The method according to claim 1 or 3, wherein the pH of the saccharification system is 4.2-4.4 and the saccharification temperature is 58-62 ℃.
5. The method according to claim 2, wherein the saccharifying enzyme is added in an amount of 2.4U/kgDS to 10U/kgDS, the pullulanase is added in an amount of 0.1U/kgDS to 0.2U/kgDS, and the acid keratinase is added in an amount of 10 to 20U/kgDS.
6. The method according to claim 5, wherein the enzymolysis condition of the protease is 30-55 ℃ and the enzymolysis time is 6-16 h.
7. The method according to claim 1, wherein the ion exchange is carried out by using cation exchange resin D001 and anion exchange resin D301.
8. The method of claim 1, comprising the steps of:
(1) saccharification: regulating the solid content of the trehalose mother liquor to 25-30, heating to 58-62 ℃, regulating the pH value to 4.2-4.4, adding saccharifying enzyme according to the ratio of 2.4U/kg DS-10U/kg DS, adding pullulanase according to the ratio of 0.1U/kg DS-0.2U/kg DS, and saccharifying for 32-72 h;
(2) cooling the saccharified liquid obtained in the step (1) to 30-55 ℃, adding acid keratinase according to 10-20U/kgDS, and reacting for 6-16h on the basis of low-speed stirring;
(3) heating the reaction liquid obtained in the step (2) to inactivate enzyme, adding activated carbon to decolorize, and filtering until the light transmittance is greater than 95%;
(4) purifying the filtrate obtained in the step (3) by adopting cation exchange resin D001 and anion exchange resin D301 in sequence, and concentrating the mass of the sugar solution to 71-73% by adopting a vacuum concentrator;
(5) adding crystal trehalose seed crystals accounting for 0.5-1.5% of the mass of the concentrated solution in the step (4), gradually reducing the temperature from 50 ℃ to 20 ℃, and using for 64-72 hours;
(6) centrifugal drying: drying in an oven at 89-90 ℃ for 4-6h after centrifugation.
9. Trehalose produced by the method of any one of claims 1 to 8.
10. Use of the method of any one of claims 1 to 8 for the production of trehalose and derivatives thereof in the fields of food, medicine and chemical industry.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0670326A2 (en) * | 1994-03-01 | 1995-09-06 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Crystalline maltosyl glucoside, and its production and use |
| CN103450288A (en) * | 2013-08-16 | 2013-12-18 | 齐鲁工业大学 | Separation and purification method of trehalose |
| WO2015089441A1 (en) * | 2013-12-13 | 2015-06-18 | Danisco Us Inc. | Serine proteases of bacillus species |
-
2019
- 2019-11-12 CN CN201911105271.2A patent/CN110818753B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0670326A2 (en) * | 1994-03-01 | 1995-09-06 | Kabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo | Crystalline maltosyl glucoside, and its production and use |
| CN103450288A (en) * | 2013-08-16 | 2013-12-18 | 齐鲁工业大学 | Separation and purification method of trehalose |
| WO2015089441A1 (en) * | 2013-12-13 | 2015-06-18 | Danisco Us Inc. | Serine proteases of bacillus species |
Non-Patent Citations (3)
| Title |
|---|
| 刘建龙等: ""海藻糖提取纯化的研究"", 《山东食品发酵》 * |
| 姜锡瑞等主编: "《生物发酵产业技术》", 31 May 2016, 中国轻工业出版社 * |
| 曹珊等: ""一种从海藻糖母液中回收海藻糖方法"", 《化工设计通讯》 * |
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Denomination of invention: A recycling method of mother liquor of crystalline Trehalose Effective date of registration: 20230711 Granted publication date: 20210528 Pledgee: Agricultural Bank of China Limited by Share Ltd. Leiyang branch Pledgor: Hunan Jindai Technology Development Co.,Ltd. Registration number: Y2023980048075 |