CN116355027B - Process for preparing glucosamine by fermentation method - Google Patents
Process for preparing glucosamine by fermentation method Download PDFInfo
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- CN116355027B CN116355027B CN202310384548.XA CN202310384548A CN116355027B CN 116355027 B CN116355027 B CN 116355027B CN 202310384548 A CN202310384548 A CN 202310384548A CN 116355027 B CN116355027 B CN 116355027B
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- glucosamine
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- decoloring
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- fermentation
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- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 title claims abstract description 69
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 title claims abstract description 69
- 229960002442 glucosamine Drugs 0.000 title claims abstract description 69
- 238000000855 fermentation Methods 0.000 title claims abstract description 50
- 230000004151 fermentation Effects 0.000 title claims abstract description 50
- 238000000034 method Methods 0.000 title claims abstract description 30
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 26
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims abstract description 56
- 239000000413 hydrolysate Substances 0.000 claims abstract description 45
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 34
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 34
- 239000013078 crystal Substances 0.000 claims abstract description 29
- 238000010438 heat treatment Methods 0.000 claims abstract description 29
- 239000007788 liquid Substances 0.000 claims abstract description 29
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims abstract description 28
- 229910021529 ammonia Inorganic materials 0.000 claims abstract description 28
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 28
- 238000001914 filtration Methods 0.000 claims abstract description 24
- 239000006228 supernatant Substances 0.000 claims abstract description 21
- 239000000706 filtrate Substances 0.000 claims abstract description 18
- 108010009736 Protein Hydrolysates Proteins 0.000 claims abstract description 15
- 239000001569 carbon dioxide Substances 0.000 claims abstract description 14
- 229910002092 carbon dioxide Inorganic materials 0.000 claims abstract description 14
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 11
- 238000001035 drying Methods 0.000 claims abstract description 5
- 239000003513 alkali Substances 0.000 claims abstract description 4
- 238000005189 flocculation Methods 0.000 claims abstract description 4
- 230000016615 flocculation Effects 0.000 claims abstract description 4
- 238000006386 neutralization reaction Methods 0.000 claims abstract description 3
- 238000001556 precipitation Methods 0.000 claims abstract description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 28
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 8
- 238000001291 vacuum drying Methods 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 6
- 230000006698 induction Effects 0.000 claims description 6
- 229920002401 polyacrylamide Polymers 0.000 claims description 6
- 229920001495 poly(sodium acrylate) polymer Polymers 0.000 claims description 4
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 4
- NNMHYFLPFNGQFZ-UHFFFAOYSA-M sodium polyacrylate Chemical compound [Na+].[O-]C(=O)C=C NNMHYFLPFNGQFZ-UHFFFAOYSA-M 0.000 claims description 4
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 3
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 2
- 230000035484 reaction time Effects 0.000 claims description 2
- 239000011347 resin Substances 0.000 claims description 2
- 229920005989 resin Polymers 0.000 claims description 2
- 238000001179 sorption measurement Methods 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims 2
- 239000008394 flocculating agent Substances 0.000 claims 1
- 238000003756 stirring Methods 0.000 description 12
- 230000007062 hydrolysis Effects 0.000 description 8
- 230000008569 process Effects 0.000 description 8
- 238000001816 cooling Methods 0.000 description 6
- 238000004042 decolorization Methods 0.000 description 6
- 238000001212 derivatisation Methods 0.000 description 6
- 150000002337 glycosamines Chemical class 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 241000238557 Decapoda Species 0.000 description 3
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 3
- 238000005903 acid hydrolysis reaction Methods 0.000 description 3
- 229910001424 calcium ion Inorganic materials 0.000 description 3
- 229910001425 magnesium ion Inorganic materials 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000012869 ethanol precipitation Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 1
- 206010002198 Anaphylactic reaction Diseases 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 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 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- 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 1
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 1
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 1
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 1
- 206010057178 Osteoarthropathies Diseases 0.000 description 1
- 241000207961 Sesamum Species 0.000 description 1
- 235000003434 Sesamum indicum Nutrition 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 208000003455 anaphylaxis Diseases 0.000 description 1
- 210000001188 articular cartilage Anatomy 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 210000001612 chondrocyte Anatomy 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229940097043 glucuronic acid Drugs 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000003303 reheating Methods 0.000 description 1
- 230000009469 supplementation Effects 0.000 description 1
Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H5/00—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium
- C07H5/04—Compounds containing saccharide radicals in which the hetero bonds to oxygen have been replaced by the same number of hetero bonds to halogen, nitrogen, sulfur, selenium, or tellurium to nitrogen
- C07H5/06—Aminosugars
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Separation Of Suspended Particles By Flocculating Agents (AREA)
Abstract
The invention discloses a process for preparing glucosamine by a fermentation method, which comprises the following steps: (1) Adding flocculant into ammonia sugar fermentation liquor for flocculation treatment, filtering after completion, and collecting filtrate for later use. (2) Adding a decoloring agent into the filtrate for decoloring, removing the decoloring agent, heating and concentrating the obtained decoloring liquid, centrifuging the obtained decoloring liquid, and collecting supernatant for later use. (3) Hydrochloric acid is added into the supernatant to carry out hydrolysis reaction under the heating condition, and alkali is added for neutralization after completion, so that hydrolysate is obtained for standby. (4) Introducing carbon dioxide into the hydrolysate, adding ethanol after completion, applying an alternating magnetic field, filtering out crystals after the precipitation of the crystals, and drying the crystals to obtain the glucosamine. The process for preparing the glucosamine by the fermentation method can extract the glucosamine from the glucosamine fermentation liquid more efficiently, and improve the yield of the glucosamine.
Description
Technical Field
The invention relates to the technical field of glucosamine preparation, in particular to a process for preparing glucosamine by a fermentation method.
Background
Glucosamine is a monosaccharide component of glucose with one hydroxyl group replaced by amino group, can be used for treating osteoarthropathy, is widely existed in human body, and is an important nutrient substance for forming chondrocyte. Glucosamine participates in the metabolism of sugar in the body, and forms important products with biological activity of the body such as hyaluronic acid, keratinocyte sulfuric acid and the like by combining with other substances (such as galactose, glucuronic acid and the like), thereby participating in the protection of the body. With age, the lack of glucosamine in the human body becomes more and more severe, and articular cartilage is continually degraded and worn. Therefore, supplementation with glucosamine by administration is an important way. At present, the main preparation modes of the aminosugar mainly comprise an acid hydrolysis method, an enzymolysis method and a microbial fermentation method. The production raw materials of the acid hydrolysis method and the enzymolysis method are basically derived from the exoskeletons of the shrimps and the crabs, namely, the chitin and the chitosan are extracted from the shrimp and the crab shells, and then the ammonia sugar is obtained through acidolysis or enzymolysis. Compared with an acid hydrolysis method and an enzymolysis method, the sesame candy prepared by the microbial fermentation method eliminates the limitation of regional seasons on raw material sources, has no fishy smell and anaphylactic reaction, and has the advantages of short production period, high strength, less environmental pollution and the like.
Disclosure of Invention
The invention provides a process for preparing glucosamine by a fermentation method, which can extract the glucosamine from a glucosamine fermentation liquid more efficiently and improve the yield of the glucosamine. In order to achieve the above purpose, the present invention discloses the following technical solutions:
a process for preparing glucosamine by a fermentation method comprises the following steps:
(1) Adding flocculant into ammonia sugar fermentation liquor for flocculation treatment, filtering after completion, and collecting filtrate for later use.
(2) Adding a decoloring agent into the filtrate for decoloring, removing the decoloring agent, heating and concentrating the obtained decoloring liquid, centrifuging the obtained decoloring liquid, and collecting supernatant for later use.
(3) Hydrochloric acid is added into the supernatant to carry out hydrolysis reaction under the heating condition, and alkali is added for neutralization after completion, so that hydrolysate is obtained for standby.
(4) Introducing carbon dioxide into the hydrolysate, applying an alternating magnetic field, adding ethanol after completion, filtering out crystals after the precipitation of the crystals, and drying the crystals to obtain the glucosamine.
Further, in the step (1), the flocculant includes any one of sodium polyacrylate, polyacrylamide, and the like. Optionally, the mass fraction of the flocculant in the ammonia sugar fermentation liquor is 0.25-0.4%.
Further, in the step (2), the decoloring agent includes any one of activated carbon, an adsorption resin, and the like. Optionally, the addition amount of the decoloring agent is 3-6 g/L.
Further, in the step (2), the time of the decoloring treatment is 15-20 min, and the decoloring treatment is performed under the water bath heating condition of 45-60 ℃.
In the step (2), the decolorized solution is heated and concentrated to 40 to 55 percent of the initial volume at the temperature of 70 to 85 ℃.
Further, in the step (2), the centrifugal speed of the centrifugal treatment is 10000-15000 r/min, and the treatment time is 10-15 min.
Further, in the step (3), the mass fraction of hydrochloric acid in the supernatant is 30-35%, the heating temperature is 80-90 ℃, and the reaction time is 4-5.5 hours, so that the N-acetylglucosamine in the supernatant is hydrolyzed into glucosamine.
Further, in the step (3), the base includes any one of sodium hydroxide, sodium carbonate, sodium bicarbonate, and the like.
Further, in the step (4), carbon dioxide is continuously introduced into the hydrolysate at a rate of 1.5-2L/min for 10-15 min. The carbon dioxide is used for forming calcium ions and magnesium ions in the hydrolysate into precipitate.
Further, in the step (4), the frequency of the alternating magnetic field is 35-50 Hz, and the magnetic induction intensity is 0.6-1.0T.
Further, in the step (4), the volume of the ethanol is 3.5-5 times of the volume of the hydrolysate. Preferably, the mass fraction of the ethanol is not less than 95%.
Further, in the step (4), the drying mode includes any one of freeze-drying, vacuum drying and the like.
Compared with the prior art, the invention has the following beneficial technical effects:
the process of the invention carries out flocculation and decoloration treatment on the ammonia sugar fermentation liquor, and then carries out heating concentration treatment, thereby not only removing redundant liquid phase in the fermentation liquor, being convenient for improving the extraction rate of the ammonia sugar and reducing the residue. And proteins and residual thalli and the like in the fermentation liquor are heated and deactivated and then settled in the process of reheating and concentration, and impurities can be further removed after centrifugal treatment, thereby being beneficial to improving the purity of the obtained amino sugar product. In the process of extracting the amino sugar component from the hydrolysate, carbon dioxide is introduced into the hydrolysate, an alternating magnetic field is applied, and ethanol is added after completion of the process to carry out ethanol precipitation. In the process, calcium and magnesium ions which remain in the ammonia sugar fermentation liquid and are provided by the culture medium react with carbon dioxide to form crystals to be separated out, meanwhile, the alternating magnetic field can effectively inhibit aggregation growth of the crystals in the forming process, so that a large number of tiny heterogeneous crystal nuclei are dispersed in the hydrolysate after treatment is finished, and when ammonia sugar in ethanol is added to be separated out, due to the existence of the heterogeneous crystal nuclei, the ammonia sugar in the hydrolysate can be more thoroughly separated out, the yield of the ammonia sugar in the hydrolysate is improved, and the waste of target products is reduced.
Detailed Description
It should be noted that the following detailed description is illustrative and is intended to provide further explanation of the invention. The invention will now be further illustrated by means of a specific implementation.
Example 1
A process for preparing glucosamine by a fermentation method comprises the following steps:
(1) Adding polyacrylamide into the ammonia sugar fermentation liquor according to the mass fraction of the flocculant in the ammonia sugar fermentation liquor of 0.35%, then rapidly and uniformly stirring, stopping stirring after floccules are formed, standing until the floccules are completely precipitated, filtering to remove the floccules, and collecting filtrate for later use.
(2) And adding an activated carbon decoloring agent into the filtrate, wherein the adding amount of the activated carbon decoloring agent is 5g/L. Then heating to 50 ℃ in water bath and preserving heat for 20min to perform decolorization treatment. Filtering to remove the decoloring agent after completion, heating and concentrating the obtained decoloring liquid to 50% of the initial volume at 80 ℃ to obtain concentrated liquid, cooling the concentrated liquid to room temperature, and performing centrifugal treatment at a centrifugal speed of 12000 r/min for 10min. After completion, the supernatant was collected for use.
(3) Adding hydrochloric acid into the supernatant until the mass fraction reaches 35%, heating to 85 ℃ for hydrolysis for 5 hours, and adding sodium hydroxide to neutralize excess hydrochloric acid after completion of the hydrolysis to obtain hydrolysate for later use.
(4) Carbon dioxide was continuously introduced into the hydrolysate at a rate of 1.5L/min for 15min, and an alternating magnetic field (frequency: 40Hz, magnetic induction strength: 0.8T) was simultaneously applied. After the completion, ethanol (mass fraction: 95%) was added to the hydrolysate in an amount of 4 times the volume of the hydrolysate, followed by standing for 40 minutes to precipitate crystals therein. And filtering out the crystal, and vacuum drying at 60 ℃ for 1 hour to obtain the glucosamine.
The yield of glucosamine in the hydrolysate of this example was tested by RP-HPL derivatization, and was = (content of glucosamine extracted-content of glucosamine in hydrolysate)/content of glucosamine in hydrolysate, yielding a yield of 96.71%.
Example 2
A process for preparing glucosamine by a fermentation method comprises the following steps:
(1) Adding sodium polyacrylate into the ammonia sugar fermentation liquor according to the mass fraction of the flocculant in the ammonia sugar fermentation liquor of 0.4%, then rapidly and uniformly stirring, stopping stirring after floccules are formed, standing until floccules are completely precipitated, filtering to remove the floccules, and collecting filtrate for later use.
(2) And adding an activated carbon decoloring agent into the filtrate, wherein the adding amount of the activated carbon decoloring agent is 3g/L. Then heating to 45 ℃ in water bath and preserving heat for 20min to perform decolorization treatment. Filtering to remove the decoloring agent after completion, heating and concentrating the obtained decoloring liquid to 55% of the initial volume at 70 ℃ to obtain concentrated liquid, cooling the concentrated liquid to room temperature, and performing centrifugal treatment at a centrifugal speed of 10000 revolutions per minute for 15 minutes. After completion, the supernatant was collected for use.
(3) Adding hydrochloric acid into the supernatant until the mass fraction reaches 30%, heating to 80 ℃ for hydrolysis for 4 hours, and adding sodium carbonate to neutralize excess hydrochloric acid after completion to obtain hydrolysate for later use.
(4) Carbon dioxide was continuously introduced into the hydrolysate at a rate of 2L/min for 10min, and an alternating magnetic field (frequency: 35Hz, magnetic induction strength: 1.0T) was simultaneously applied. After the completion, ethanol (mass fraction: 99%) was added in an amount of 5 times the volume of the hydrolysate, and then the mixture was left to stand for 30 minutes to precipitate crystals therein. And filtering out the crystal, and vacuum drying at 60 ℃ for 1 hour to obtain the glucosamine.
The yield of glucosamine in the hydrolysate of this example was tested by RP-HPL derivatization, and the yield= (content of glucosamine extracted-content of glucosamine in the hydrolysate)/content of glucosamine in the hydrolysate, giving a yield of 94.87%.
Example 3
A process for preparing glucosamine by a fermentation method comprises the following steps:
(1) Adding polyacrylamide into the ammonia sugar fermentation liquor according to the mass fraction of the flocculant in the ammonia sugar fermentation liquor of 0.25%, then rapidly and uniformly stirring, stopping stirring after floccules are formed, standing until the floccules are completely precipitated, filtering to remove the floccules, and collecting filtrate for later use.
(2) And adding an activated carbon decoloring agent into the filtrate, wherein the adding amount of the activated carbon decoloring agent is 6g/L. Then heating to 60 ℃ in water bath and preserving heat for 15min to carry out decolorization treatment. Filtering to remove the decoloring agent after completion, heating and concentrating the obtained decoloring liquid to 40% of the initial volume at 85 ℃ to obtain concentrated liquid, cooling the concentrated liquid to room temperature, and performing centrifugal treatment at a centrifugal speed of 15000 r/min for 10min. After completion, the supernatant was collected for use.
(3) Adding hydrochloric acid into the supernatant until the mass fraction reaches 35%, heating to 90 ℃ for hydrolysis for 5.5 hours, and adding sodium bicarbonate to neutralize excess hydrochloric acid after completion to obtain hydrolysate for later use.
(4) Carbon dioxide was continuously introduced into the hydrolysate at a rate of 1.8L/min for 12min, and an alternating magnetic field (frequency: 50Hz, magnetic induction strength: 0.6T) was simultaneously applied. After the completion, ethanol (mass fraction: 99%) 3.5 times the volume of the hydrolysate was added, followed by standing for 40 minutes to precipitate crystals therein. And filtering out the crystal, and vacuum drying at 60 ℃ for 1 hour to obtain the glucosamine.
The yield of glucosamine in the hydrolysate of this example was tested by RP-HPL derivatization, and was = (content of glucosamine extracted-content of glucosamine in hydrolysate)/content of glucosamine in hydrolysate, yielding a yield of 96.26%.
Example 4
A process for preparing glucosamine by a fermentation method comprises the following steps:
(1) Adding polyacrylamide into the ammonia sugar fermentation liquor according to the mass fraction of the flocculant in the ammonia sugar fermentation liquor of 0.35%, then rapidly and uniformly stirring, stopping stirring after floccules are formed, standing until the floccules are completely precipitated, filtering to remove the floccules, and collecting filtrate for later use.
(2) And adding an activated carbon decoloring agent into the filtrate, wherein the adding amount of the activated carbon decoloring agent is 5g/L. Then heating to 50 ℃ in water bath and preserving heat for 20min to perform decolorization treatment. Filtering to remove the decoloring agent after completion, heating and concentrating the obtained decoloring liquid to 50% of the initial volume at 80 ℃ to obtain concentrated liquid, cooling the concentrated liquid to room temperature, and performing centrifugal treatment at a centrifugal speed of 12000 r/min for 10min. After completion, the supernatant was collected for use.
(3) Adding hydrochloric acid into the supernatant until the mass fraction reaches 35%, heating to 85 ℃ for hydrolysis for 5 hours, and adding sodium hydroxide to neutralize excess hydrochloric acid after completion of the hydrolysis to obtain hydrolysate for later use.
(4) And (3) applying an alternating magnetic field (frequency is 40Hz and magnetic induction intensity is 0.8T) to the hydrolysate for 15min, adding ethanol (mass fraction is 95%) which is 4 times the volume of the hydrolysate after the completion, and standing for 40min to precipitate crystals in the hydrolysate. And filtering out the crystal, and vacuum drying at 60 ℃ for 1 hour to obtain the glucosamine.
The yield of glucosamine in the hydrolysate of this example was tested by RP-HPL derivatization, and was = (content of glucosamine extracted-content of glucosamine in hydrolysate)/content of glucosamine in hydrolysate, yielding a yield of 87.43%.
Example 5
A process for preparing glucosamine by a fermentation method comprises the following steps:
(1) Adding sodium polyacrylate into the ammonia sugar fermentation liquor according to the mass fraction of the flocculant in the ammonia sugar fermentation liquor of 0.4%, then rapidly and uniformly stirring, stopping stirring after floccules are formed, standing until floccules are completely precipitated, filtering to remove the floccules, and collecting filtrate for later use.
(2) And adding an activated carbon decoloring agent into the filtrate, wherein the adding amount of the activated carbon decoloring agent is 3g/L. Then heating to 45 ℃ in water bath and preserving heat for 20min to perform decolorization treatment. Filtering to remove the decoloring agent after completion, heating and concentrating the obtained decoloring liquid to 55% of the initial volume at 70 ℃ to obtain concentrated liquid, cooling the concentrated liquid to room temperature, and performing centrifugal treatment at a centrifugal speed of 10000 revolutions per minute for 15 minutes. After completion, the supernatant was collected for use.
(3) Adding hydrochloric acid into the supernatant until the mass fraction reaches 30%, heating to 80 ℃ for hydrolysis for 4 hours, and adding sodium carbonate to neutralize excess hydrochloric acid after completion to obtain hydrolysate for later use.
(4) Carbon dioxide was continuously introduced into the hydrolysate at a rate of 2L/min for 10min. After the completion, ethanol (mass fraction: 99%) was added in an amount of 5 times the volume of the hydrolysate, and then the mixture was left to stand for 30 minutes to precipitate crystals therein. And filtering out the crystal, and vacuum drying at 60 ℃ for 1 hour to obtain the glucosamine.
The yield of glucosamine in the hydrolysate of this example was tested by RP-HPL derivatization, and was = (content of glucosamine extracted-content of glucosamine in hydrolysate)/content of glucosamine in hydrolysate, yielding a yield of 89.57%.
Example 6
A process for preparing glucosamine by a fermentation method comprises the following steps:
(1) Adding polyacrylamide into the ammonia sugar fermentation liquor according to the mass fraction of the flocculant in the ammonia sugar fermentation liquor of 0.25%, then rapidly and uniformly stirring, stopping stirring after floccules are formed, standing until the floccules are completely precipitated, filtering to remove the floccules, and collecting filtrate for later use.
(2) And adding an activated carbon decoloring agent into the filtrate, wherein the adding amount of the activated carbon decoloring agent is 6g/L. Then heating to 60 ℃ in water bath and preserving heat for 15min to carry out decolorization treatment. Filtering to remove the decoloring agent after completion, heating and concentrating the obtained decoloring liquid to 40% of the initial volume at 85 ℃ to obtain concentrated liquid, cooling the concentrated liquid to room temperature, and performing centrifugal treatment at a centrifugal speed of 15000 r/min for 10min. After completion, the supernatant was collected for use.
(3) Adding hydrochloric acid into the supernatant until the mass fraction reaches 35%, heating to 90 ℃ for hydrolysis for 5.5 hours, and adding sodium bicarbonate to neutralize excess hydrochloric acid after completion to obtain hydrolysate for later use.
(4) Ethanol (99% by mass) was added to the hydrolysate in an amount 3.5 times the volume of the hydrolysate, and the mixture was allowed to stand for 40 minutes to precipitate crystals therein. And filtering out the crystal, and vacuum drying at 60 ℃ for 1 hour to obtain the glucosamine.
The yield of glucosamine in the hydrolysate of this example was tested by RP-HPL derivatization, and was = (content of glucosamine extracted-content of glucosamine in hydrolysate)/content of glucosamine in hydrolysate, yielding a yield of 81.93%.
It can be seen that the yields of examples 1, 2 and 3 are higher than those of examples 4, 5 and 6, mainly because examples 1 to 3 were obtained by subjecting the hydrolysate to carbon dioxide and an alternating magnetic field, and then adding ethanol to the hydrolysate to effect ethanol precipitation in the step of extracting the amino sugar component from the hydrolysate. In the process, calcium and magnesium ions which remain in the ammonia sugar fermentation liquid and are provided by the culture medium react with carbon dioxide to form crystals to be separated out, meanwhile, the alternating magnetic field can effectively inhibit aggregation growth of the crystals in the forming process, so that a large number of tiny heterogeneous crystal nuclei are dispersed in the hydrolysate after treatment is finished, and when ammonia sugar in ethanol is added to be separated out, due to the existence of the heterogeneous crystal nuclei, the ammonia sugar in the hydrolysate can be more thoroughly separated out, the yield of the ammonia sugar in the hydrolysate is improved, and the waste of target products is reduced.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (11)
1. A process for preparing glucosamine by a fermentation method is characterized by comprising the following steps:
(1) Adding a flocculating agent into the ammonia sugar fermentation liquor for flocculation treatment, filtering after completion, and collecting filtrate for later use;
(2) Adding a decoloring agent into the filtrate for decoloring, removing the decoloring agent, heating and concentrating the obtained decoloring liquid, centrifuging the obtained decoloring liquid, and collecting supernatant for later use;
(3) Hydrochloric acid is added into the supernatant to carry out hydrolysis reaction under the heating condition, and alkali is added for neutralization after completion, so as to obtain hydrolysate for standby;
(4) Introducing carbon dioxide into the hydrolysate, applying an alternating magnetic field, adding ethanol after completion, filtering out crystals after the precipitation of the crystals, and drying the crystals to obtain glucosamine;
in the step (3), the mass fraction of hydrochloric acid in the supernatant is 30-35%, the heating temperature is 80-90 ℃, and the reaction time is 4-5.5 hours;
in the step (3), the alkali comprises any one of sodium hydroxide, sodium carbonate and sodium bicarbonate;
in the step (4), continuously introducing carbon dioxide into the hydrolysate at a rate of 1.5-2L/min for 10-15 min;
in the step (4), the frequency of the alternating magnetic field is 35-50 Hz, and the magnetic induction intensity is 0.6-1.0T.
2. The process for preparing glucosamine by fermentation according to claim 1, wherein in the step (1), the flocculant comprises any one of sodium polyacrylate and polyacrylamide.
3. The process for preparing glucosamine by fermentation according to claim 1, wherein the mass fraction of the flocculant in the ammonia sugar fermentation liquid is 0.25-0.4%.
4. The process for preparing glucosamine by fermentation according to claim 1, wherein in the step (2), the decoloring agent comprises any one of activated carbon and an adsorption resin.
5. The process for preparing glucosamine by fermentation according to claim 4, wherein the decoloring agent is added in an amount of 3 to 6g/L.
6. The process for preparing glucosamine by fermentation according to claim 1, wherein in the step (2), the decoloring treatment is performed for 15 to 20 minutes under a water bath heating condition of 45 to 60 ℃.
7. The process for preparing glucosamine by fermentation according to claim 1, wherein in the step (2), the decolorized solution is heated and concentrated to 40-55% of the initial volume at 70-85 ℃.
8. The process for preparing glucosamine by fermentation according to claim 1, wherein in the step (2), the centrifugation rate of the centrifugation is 10000-15000 rpm and the treatment time is 10-15 min.
9. The process for preparing glucosamine by fermentation according to claim 1, wherein in the step (4), the volume of the ethanol is 3.5 to 5 times the volume of the hydrolysate.
10. The process for preparing glucosamine by fermentation according to claim 9, wherein the mass fraction of the ethanol is not less than 95%.
11. The process for preparing glucosamine by fermentation according to any one of claims 1-10, wherein in step (4), the drying means comprises any one of lyophilization and vacuum drying.
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| CN115340580A (en) * | 2021-05-14 | 2022-11-15 | 山东润德生物科技有限公司 | Preparation method of glucosamine sulfate and sodium chloride double salt |
| CN115838389A (en) * | 2022-11-07 | 2023-03-24 | 山东润德生物科技有限公司 | Method for improving extraction rate of glucosamine in fermentation liquor |
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| CN115340580A (en) * | 2021-05-14 | 2022-11-15 | 山东润德生物科技有限公司 | Preparation method of glucosamine sulfate and sodium chloride double salt |
| CN115838389A (en) * | 2022-11-07 | 2023-03-24 | 山东润德生物科技有限公司 | Method for improving extraction rate of glucosamine in fermentation liquor |
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