CN117286194A - Process method for co-producing glucuronic acid and D-chiro-inositol - Google Patents
Process method for co-producing glucuronic acid and D-chiro-inositol Download PDFInfo
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- CN117286194A CN117286194A CN202311187963.2A CN202311187963A CN117286194A CN 117286194 A CN117286194 A CN 117286194A CN 202311187963 A CN202311187963 A CN 202311187963A CN 117286194 A CN117286194 A CN 117286194A
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- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 title claims abstract description 114
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 title claims abstract description 44
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 title claims abstract description 44
- 229940097043 glucuronic acid Drugs 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 28
- 230000008569 process Effects 0.000 title claims abstract description 18
- CDAISMWEOUEBRE-LKPKBOIGSA-N 1D-chiro-inositol Chemical compound O[C@H]1[C@@H](O)[C@H](O)[C@H](O)[C@@H](O)[C@@H]1O CDAISMWEOUEBRE-LKPKBOIGSA-N 0.000 title claims description 44
- 229960000367 inositol Drugs 0.000 claims abstract description 70
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 claims abstract description 68
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 claims abstract description 61
- 239000003957 anion exchange resin Substances 0.000 claims abstract description 14
- 101710088194 Dehydrogenase Proteins 0.000 claims abstract description 13
- 102000004316 Oxidoreductases Human genes 0.000 claims abstract description 9
- 108090000854 Oxidoreductases Proteins 0.000 claims abstract description 9
- 239000003054 catalyst Substances 0.000 claims abstract description 9
- 102000004190 Enzymes Human genes 0.000 claims abstract description 8
- 108090000790 Enzymes Proteins 0.000 claims abstract description 8
- 102000004195 Isomerases Human genes 0.000 claims abstract description 8
- 108090000769 Isomerases Proteins 0.000 claims abstract description 8
- 238000001035 drying Methods 0.000 claims abstract description 6
- 239000000843 powder Substances 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 50
- 239000012528 membrane Substances 0.000 claims description 45
- 239000007788 liquid Substances 0.000 claims description 42
- 238000006243 chemical reaction Methods 0.000 claims description 25
- 238000000108 ultra-filtration Methods 0.000 claims description 22
- 238000001914 filtration Methods 0.000 claims description 17
- 239000000919 ceramic Substances 0.000 claims description 16
- 238000005406 washing Methods 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- 239000000706 filtrate Substances 0.000 claims description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- 238000001816 cooling Methods 0.000 claims description 7
- 239000008055 phosphate buffer solution Substances 0.000 claims description 7
- 230000000694 effects Effects 0.000 claims description 6
- 238000001694 spray drying Methods 0.000 claims description 6
- 239000012141 concentrate Substances 0.000 claims description 5
- 239000008367 deionised water Substances 0.000 claims description 5
- 229910021641 deionized water Inorganic materials 0.000 claims description 5
- 238000000967 suction filtration Methods 0.000 claims description 5
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 claims description 4
- 239000003729 cation exchange resin Substances 0.000 claims description 4
- 208000028659 discharge Diseases 0.000 claims description 4
- 238000011033 desalting Methods 0.000 claims description 2
- 239000011148 porous material Substances 0.000 claims description 2
- 238000000746 purification Methods 0.000 abstract description 5
- 238000002425 crystallisation Methods 0.000 abstract description 4
- 230000008025 crystallization Effects 0.000 abstract description 4
- 238000000926 separation method Methods 0.000 abstract description 4
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 239000011347 resin Substances 0.000 description 12
- 229920005989 resin Polymers 0.000 description 12
- 125000000129 anionic group Chemical group 0.000 description 6
- 239000012530 fluid Substances 0.000 description 6
- 230000035484 reaction time Effects 0.000 description 6
- 239000000047 product Substances 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 125000002091 cationic group Chemical group 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 238000007599 discharging Methods 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 210000000653 nervous system Anatomy 0.000 description 3
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 230000019491 signal transduction Effects 0.000 description 2
- 102000018711 Facilitative Glucose Transport Proteins Human genes 0.000 description 1
- 108091052347 Glucose transporter family Proteins 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 230000005017 genetic modification Effects 0.000 description 1
- 235000013617 genetically modified food Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000004165 myocardium Anatomy 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 201000010065 polycystic ovary syndrome Diseases 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
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- 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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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
- C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
- C07C29/76—Separation; Purification; Use of additives, e.g. for stabilisation by physical treatment
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- C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
- C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
- C07C29/76—Separation; Purification; Use of additives, e.g. for stabilisation by physical treatment
- C07C29/78—Separation; Purification; Use of additives, e.g. for stabilisation by physical treatment by condensation or crystallisation
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- 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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- C12P7/00—Preparation of oxygen-containing organic compounds
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Abstract
The invention discloses a process method for co-producing glucuronic acid and D-chiral inositol, which relates to the technical field of D-chiral inositol production, and adopts mixed enzyme of inositol dehydrogenase and inositol monoketone isomerase and whole-cell catalyst containing inositol oxidase to convert inositol into mixed liquor of glucuronic acid and D-chiral inositol, wherein the glucuronic acid is adsorbed by an anion exchange resin column, so that the purification of D-chiral inositol is realized, the separation is simple and thorough, the separation cost is greatly reduced, and finally D-chiral inositol and glucuronic acid dry powder are obtained through concentration crystallization and drying respectively, and the product has high purity and good quality.
Description
Technical Field
The invention relates to the technical field of D-chiro-inositol production, in particular to a process method for co-producing glucuronic acid and D-chiro-inositol.
Background
D-chiro-inositol is widely present in organisms, particularly in the nervous system and cardiac muscle, at high levels. It is an important secondary messenger molecule that can be involved in a variety of biological metabolism and signal transduction processes. In humans, the main functions of D-chiro-inositol include the following:
promote the absorption and utilization of glucose: d-chiro-inositol can increase the expression and activity of glucose transporter, promote the absorption and utilization of glucose by cells, and thus maintain normal blood glucose level.
Regulate the structure and function of cell membranes: d-chiro-inositol can bind to cell membranes, regulate the fluidity and stability of the membranes, and affect the signal transduction of the cell membranes and the permeability of the molecular channels.
Improving the function of the nervous system: d-chiro-inositol can promote the growth and regeneration of neurons, increase the survival rate and activity of nerve cells, and is helpful for improving the functions of the nervous system.
Antioxidant effect: the D-chiro-inositol has strong antioxidation, can neutralize free radicals, reduce oxidative damage and protect cells from damage.
In conclusion, the D-chiro-inositol can be applied to a plurality of fields such as health care products, cosmetics and the like, and has wide application prospect.
The chiral inositol is synthesized by a chemical synthesis method, breakthrough is not realized at present, the process steps are complex, and the cost is high. Yoshida et al (Yoshida K, et al Genetic modification of Bacillus subtilis forproduction of D-chiro-inolitol, an investigational drug candidate fortreatment of type 2diabetes and polycystic ovary syndrome.Appl EnvironMicrobiol 72:1310-1315,2006.) utilize genetically engineered bacillus subtilis to convert myo-inositol to D-chiro-inositol by fermentation, but the yield is only 6%, the conversion rate is low, and the resulting D-chiro-inositol and myo-inositol are difficult to separate, resulting in low purity of D-chiro-inositol.
Disclosure of Invention
The technical problems to be solved by the invention are as follows: aiming at the defects existing in the prior art, the process method for co-producing glucuronic acid and D-chiro-inositol is provided, and the conversion rate and purity of the product are high and the product is easy to separate.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a process method for co-producing glucuronic acid and D-chiro-inositol comprises the following steps:
a: adding mixed enzyme of inositol dehydrogenase and inositol monoketone isomerase into 10-100g/L inositol solution, adding phosphate buffer solution to regulate pH to 6.5-7.5, and converting at 35-45 deg.c for 4-10 hr to obtain converted solution I containing D-chiral inositol and inositol;
b: filtering the conversion solution I by a ceramic membrane, collecting a ceramic membrane clear solution, adding a whole cell catalyst containing inositol oxidase into the ceramic membrane clear solution, then adding a phosphate buffer solution to adjust the pH of the system to 6.5-7.5, and converting for 4-6 hours at 35-45 ℃ to obtain a conversion solution II containing D-chiral inositol and glucuronic acid;
c: filtering the conversion solution II to collect filtrate, adsorbing glucuronic acid by the filtrate through anion exchange resin, and collecting effluent;
d: analyzing the anion exchange resin by using 5-8wt% hydrochloric acid solution, and collecting analysis solution;
e: concentrating the collected effluent liquid in a concentrator under vacuum < -0.09Mpa at the water bath temperature of 50-70 ℃ until the solid content is 60-70% w/w to obtain concentrated solution I, adding 95wt% ethanol, cooling to 10-15 ℃, carrying out suction filtration, and drying to obtain D-chiral inositol;
f: concentrating the collected analytical solution in a concentrator under vacuum < -0.09Mpa at 50-70deg.C in water bath, and spray drying to obtain glucuronic acid dry powder.
Preferably, the mass ratio of inositol dehydrogenase to inositol monoketone isomerase in step a is 1: 0.5-1, wherein the enzyme activity of the inositol dehydrogenase is 30-35U/mg, the enzyme activity of the inositol monoketone isomerase is 25-30U/mg, and the volume ratio of the inositol dehydrogenase to the inositol solution is 1-5g/L.
Preferably, the ratio of the total catalyst addition containing inositol oxidase to the volume of inositol solution in step B is 20-30g/L.
Preferably, the pore diameter of the ceramic membrane in the step B is 20-100nm, and the ceramic membrane concentrate is subjected to environment-friendly pollution discharge treatment.
Preferably, the filtrate in the step C is filtered by an ultrafiltration membrane to collect an ultrafiltration membrane clear solution, the ultrafiltration membrane clear solution is subjected to anion exchange resin to adsorb glucuronic acid, and effluent liquid is collected, wherein the aperture of the ultrafiltration membrane is 1000-10000Da, and the ultrafiltration membrane concentrate is subjected to environment-friendly pollution discharge treatment.
Preferably, in the step C, the clear solution of the ultrafiltration membrane firstly enters cation exchange resin for desalting, the desalted solution enters anion exchange resin for adsorbing glucuronic acid, the flow rate of the clear solution of the ultrafiltration membrane entering the cation exchange resin is 1-2BV/h, and the flow rate of the desalted solution entering the anion exchange resin is 1-2BV/h.
Preferably, after the effluent liquid in the step C is collected, the anion exchange resin is subjected to top washing by deionized water, the top washing liquid is collected, and the effluent liquid and the top washing liquid are combined and enter a concentrator in the step E for concentration.
Preferably, the amount of 95wt% ethanol added in step E is 0.5-3.0 times the volume of concentrate I.
Preferably, the cooling rate in the step E is 1-5 ℃/h.
Preferably, the resolving liquid in the step F is concentrated to a solid content of 30-50% w/w and then spray-dried.
Due to the adoption of the technical scheme, the invention has the beneficial effects that:
1. inositol is used as the main raw material, so that the price is low, and the production cost is reduced.
2. The mixed enzyme of inositol dehydrogenase and inositol monoketoisomerase and the whole-cell catalyst containing inositol oxidase are adopted to convert inositol into mixed liquor of glucuronic acid and D-chiral inositol, wherein the glucuronic acid is adsorbed by an anion exchange resin column, so that the purification of D-chiral inositol is realized, the separation is simple and thorough, the separation cost is greatly reduced, and meanwhile, the purity of the product is high and the quality is good.
3. The D-chiro-inositol is converted into glucuronic acid, the molar ratio of the D-chiro-inositol after conversion is about 15%, the molar ratio of the glucuronic acid is about 85%, the inositol is completely converted, the conversion rate is high, and the overall economic benefit is improved by the glucuronic acid.
4. The process method of the invention has less sewage and residues and is environment-friendly.
Drawings
FIG. 1 is a liquid chromatogram of D-chiro-inositol in example 1 of the present invention;
FIG. 2 is a liquid chromatogram of glucuronic acid in example 1 of the present invention.
Detailed Description
The invention is further illustrated by the following examples.
Example 1
(1) Preparing 60L of inositol solution with the concentration of 10g/L, adding 6g of inositol dehydrogenase and 3g of inositol monoketoisomerase, adding 50mM phosphate buffer solution to regulate the pH value of a reaction system to 6.5, controlling the reaction temperature to 35 ℃ and the reaction time to 4 hours, and obtaining feed liquid containing D-chiral inositol for standby after the reaction is finished;
(2) Filtering the feed liquid containing D-chiro-inositol in the step (1) by adopting a ceramic membrane with the molecular weight cut-off of 20nm, controlling the membrane feeding pressure to be 0.3MPa, the membrane discharging pressure to be 0.2MPa, and filtering and collecting 60L of filtrate for later use;
(3) Adding 1200g of whole-cell catalyst containing inositol oxidase into the filtrate obtained in the step (2), controlling the reaction temperature to 35 ℃ and the reaction time to 4 hours, and filtering by a ceramic membrane after the reaction is finished to obtain 60L of feed liquid containing D-chiro-inositol and glucuronic acid for later use;
(4) Taking the feed liquid containing D-chiro-inositol and glucuronic acid in the step (3), filtering by an ultrafiltration membrane with the aperture of 1000Da, and collecting the clear liquid of the ultrafiltration membrane for standby;
(5) Taking the clear solution of the ultrafiltration membrane in the step (4), passing through a cationic resin column (SQD-65) at a flow rate of 1BV/h, passing the collected first effluent through an anionic resin column (D-296) at a flow rate of 1BV/h, collecting the second effluent, washing the resin column with 1BV deionized water, collecting the top washing liquid, combining the second effluent with the top washing liquid to obtain D-chiral inositol purification liquid, analyzing the top washed anionic resin column with 5wt% hydrochloric acid, and collecting the analysis liquid for later use;
(6) Concentrating the D-chiro-inositol purified solution obtained in the step (5) under vacuum < -0.09Mpa at the water bath temperature of 50 ℃ until the solid content is 60% w/w, taking 1L of concentrated solution, adding 0.5L of 95wt% ethanol, cooling to 15 ℃ at the speed of 1 ℃/h, carrying out suction filtration, and drying to obtain 537g of D-chiro-inositol, wherein the crystallization yield is 89.5% and the purity is 99.3%.
(7) Concentrating the resolving fluid in the step (5) under vacuum < -0.09Mpa at water bath temperature of 50deg.C until the solid content is 30% w/w, collecting 1L of concentrated fluid, and spray drying (inlet air temperature is 110deg.C, outlet temperature is 90deg.C, and feeding flow rate is 10 mL/min) to obtain 289g of glucuronic acid with yield of 96.3% and purity of 99.1%.
Example 2
(1) Preparing 60L of 50g/L inositol solution, adding 150g of inositol dehydrogenase and 150g of inositol monoketone isomerase, adding 50mM phosphate buffer solution to adjust the pH of a reaction system to 7.0, controlling the reaction temperature to 40 ℃ and the reaction time to 7h, and obtaining a feed liquid containing D-chiral inositol for later use after the reaction is finished;
(2) Filtering the feed liquid containing D-chiro-inositol in the step (1) by adopting a ceramic membrane with the molecular weight cut-off of 50nm, controlling the membrane feeding pressure to be 0.3MPa, the membrane discharging pressure to be 0.2MPa, and filtering and collecting 60L of filtrate for later use;
(3) Taking the filtrate in the step (2), adding 1500g of whole-cell catalyst containing inositol oxidase, controlling the reaction temperature to 40 ℃ and the reaction time to 5 hours, and filtering by a ceramic membrane after the reaction is finished to obtain 60L of feed liquid containing D-chiro-inositol and glucuronic acid for later use;
(4) Taking the feed liquid containing D-chiro-inositol and glucuronic acid in the step (3), filtering by an ultrafiltration membrane with the aperture of 5000Da, and collecting the clear liquid of the ultrafiltration membrane for later use;
(5) Taking the clear solution of the ultrafiltration membrane in the step (4), passing through a cationic resin column (SQD-65) at a flow rate of 1.5BV/h, collecting a first effluent, passing through an anionic resin column (D-296) at a flow rate of 1.5BV/h, collecting a second effluent, washing the resin column with 1.5BV deionized water, collecting a top washing liquid, combining the second effluent with the top washing liquid to obtain a D-chiral inositol purification liquid, analyzing the top washed anionic resin column with 6wt% hydrochloric acid, and collecting an analysis liquid for later use;
(6) Concentrating the D-chiro-inositol purified solution obtained in the step (5) under vacuum < -0.09Mpa at water bath temperature of 60 ℃ until the solid content is 65% w/w, taking 1L of concentrated solution, adding 1.5L of 95wt% ethanol, cooling to 12 ℃ at a speed of 3 ℃/h, carrying out suction filtration, and drying to obtain 593g of D-chiro-inositol, wherein the crystallization yield is 91.2% and the purity is 99.6%.
(7) Concentrating the resolving fluid in the step (5) under vacuum < -0.09Mpa at water bath temperature of 60 ℃ until the solid content is 40% w/w, taking 1L of concentrated fluid, and spray drying (inlet air temperature is 120 ℃, outlet temperature is 90 ℃, and feeding flow rate is 10 mL/min) to obtain 390g of glucuronic acid, wherein the yield is 97.4%, and the purity is 99.5%.
Example 3
(1) Preparing 60L of inositol solution with the concentration of 100g/L, adding 300g of inositol dehydrogenase and 200g of inositol monoketoisomerase, adding 50mM phosphate buffer solution to regulate the pH value of a reaction system to 7.5, controlling the reaction temperature to 45 ℃ and the reaction time to 10 hours, and obtaining feed liquid containing D-chiral inositol for standby after the reaction is finished;
(2) Filtering the feed liquid containing D-chiro-inositol in the step (1) by adopting a ceramic membrane with the molecular weight cut-off of 100nm, controlling the membrane feeding pressure to be 0.3MPa, the membrane discharging pressure to be 0.2MPa, and filtering and collecting 60L of filtrate for later use;
(3) Taking the filtrate in the step (2), adding 1800g of whole-cell catalyst containing inositol oxidase, controlling the reaction temperature to 45 ℃ and the reaction time to 6 hours, and filtering by a ceramic membrane after the reaction is finished to obtain 60L of feed liquid containing D-chiro-inositol and glucuronic acid for later use;
(4) Taking the feed liquid containing D-chiro-inositol and glucuronic acid in the step (3), filtering by an ultrafiltration membrane with the aperture of 10000Da, and collecting the clear liquid of the ultrafiltration membrane for standby;
(5) Taking filtrate in the step (4), passing through a cationic resin column (SQD-65) at a flow rate of 2BV/h, collecting a first effluent, passing through an anionic resin column (D-296) at a flow rate of 2BV/h, collecting a second effluent, washing the resin column with 2BV deionized water, collecting a top washing liquid, combining the second effluent with the top washing liquid to obtain D-chiral inositol purification liquid, analyzing the top washed anionic resin column with 8wt% hydrochloric acid, and collecting an analysis liquid for later use;
(6) Concentrating the D-chiro-inositol purified solution obtained in the step (5) under vacuum < -0.09Mpa at the water bath temperature of 70 ℃ until the solid content is 70% w/w, taking 1L of concentrated solution, adding 3L of 95wt% ethanol, cooling to 10 ℃ at the speed of 5 ℃/h, carrying out suction filtration, and drying to obtain 636g of D-chiro-inositol, wherein the crystallization yield is 90.8% and the purity is 99.5%.
(7) Concentrating the resolving fluid in the step (5) under vacuum < -0.09Mpa at 70 ℃ in water bath temperature to solid content of 50% w/w, taking 1L of concentrated fluid, and spray drying (inlet air temperature is 120 ℃, outlet temperature is 95 ℃, and feeding flow rate is 10 mL/min) to obtain 484g of glucuronic acid, wherein the yield is 96.8%, and the purity is 99.4%.
It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. Further, it is understood that various changes and modifications may be made by those skilled in the art after reading the teachings of the present invention, and such equivalents are intended to fall within the scope of the claims appended hereto.
Claims (10)
1. A process method for co-producing glucuronic acid and D-chiro-inositol is characterized by comprising the following steps:
a: adding mixed enzyme of inositol dehydrogenase and inositol monoketone isomerase into 10-100g/L inositol solution, adding phosphate buffer solution to regulate pH to 6.5-7.5, and converting at 35-45 deg.c for 4-10 hr to obtain converted solution I containing D-chiral inositol and inositol;
b: filtering the conversion solution I by a ceramic membrane, collecting a ceramic membrane clear solution, adding a whole cell catalyst containing inositol oxidase into the ceramic membrane clear solution, then adding a phosphate buffer solution to adjust the pH of the system to 6.5-7.5, and converting for 4-6 hours at 35-45 ℃ to obtain a conversion solution II containing D-chiral inositol and glucuronic acid;
c: filtering the conversion solution II to collect filtrate, adsorbing glucuronic acid by the filtrate through anion exchange resin, and collecting effluent;
d: analyzing the anion exchange resin by using 5-8wt% hydrochloric acid solution, and collecting analysis solution;
e: concentrating the collected effluent liquid in a concentrator under vacuum < -0.09Mpa at the water bath temperature of 50-70 ℃ until the solid content is 60-70% w/w to obtain concentrated solution I, adding 95wt% ethanol, cooling to 10-15 ℃, carrying out suction filtration, and drying to obtain D-chiral inositol;
f: concentrating the collected analytical solution in a concentrator under vacuum < -0.09Mpa at 50-70deg.C in water bath, and spray drying to obtain glucuronic acid dry powder.
2. The process for co-producing glucuronic acid and D-chiro-inositol according to claim 1, wherein: the mass ratio of inositol dehydrogenase to inositol monoketone isomerase in step A is 1: 0.5-1, wherein the enzyme activity of the inositol dehydrogenase is 30-35U/mg, the enzyme activity of the inositol monoketone isomerase is 25-30U/mg, and the volume ratio of the inositol dehydrogenase to the inositol solution is 1-5g/L.
3. The process for co-producing glucuronic acid and D-chiro-inositol according to claim 1, wherein: the volume ratio of the whole cell catalyst containing inositol oxidase to the inositol solution in the step B is 20-30g/L.
4. The process for co-producing glucuronic acid and D-chiro-inositol according to claim 1, wherein: and B, performing environment-friendly pollution discharge treatment on the ceramic membrane concentrate in the step of pore diameter of the ceramic membrane of 20-100 nm.
5. The process for co-producing glucuronic acid and D-chiro-inositol according to claim 1, wherein: and C, filtering the filtrate by an ultrafiltration membrane to collect an ultrafiltration membrane clear solution, adsorbing glucuronic acid by the ultrafiltration membrane clear solution through anion exchange resin, and collecting effluent, wherein the aperture of the ultrafiltration membrane is 1000-10000Da, and carrying out environment-friendly pollution discharge treatment on the ultrafiltration membrane concentrate.
6. The process for co-producing glucuronic acid and D-chiro-inositol according to claim 5, wherein: in the step C, the clear solution of the ultrafiltration membrane firstly enters cation exchange resin for desalting, the desalted solution enters anion exchange resin for adsorbing glucuronic acid, the flow rate of the clear solution of the ultrafiltration membrane entering the cation exchange resin is 1-2BV/h, and the flow rate of the desalted solution entering the anion exchange resin is 1-2BV/h.
7. The process for co-producing glucuronic acid and D-chiro-inositol according to claim 1, wherein: and C, after the effluent liquid is collected, washing the anion exchange resin with deionized water, collecting top washing liquid, and combining the effluent liquid and the top washing liquid into a concentrator in the step E for concentration.
8. The process for co-producing glucuronic acid and D-chiro-inositol according to claim 1, wherein: the addition amount of 95wt% ethanol in the step E is 0.5-3.0 times of the volume of the concentrated solution I.
9. The process for co-producing glucuronic acid and D-chiro-inositol according to claim 1, wherein: and E, cooling at a speed of 1-5 ℃/h.
10. The process for co-producing glucuronic acid and D-chiro-inositol according to claim 1, wherein: and F, concentrating the resolved liquid to a solid content of 30-50% w/w, and then spray drying.
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| CN202311187963.2A CN117286194A (en) | 2023-09-15 | 2023-09-15 | Process method for co-producing glucuronic acid and D-chiro-inositol |
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| CN202311187963.2A CN117286194A (en) | 2023-09-15 | 2023-09-15 | Process method for co-producing glucuronic acid and D-chiro-inositol |
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