CN110846373A - Method for producing glucosamine potassium sulfate salt by microbial fermentation - Google Patents

Method for producing glucosamine potassium sulfate salt by microbial fermentation Download PDF

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CN110846373A
CN110846373A CN201911210934.7A CN201911210934A CN110846373A CN 110846373 A CN110846373 A CN 110846373A CN 201911210934 A CN201911210934 A CN 201911210934A CN 110846373 A CN110846373 A CN 110846373A
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potassium sulfate
glucosamine
fermentation
microbial fermentation
steps
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卢健行
韩宁
吴祥舟
张建华
卢建功
卢建智
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Shandong Runde Biotechnology Co Ltd
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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
    • C12P19/26—Preparation of nitrogen-containing carbohydrates

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Abstract

The invention discloses a method for producing glucosamine potassium sulfate by microbial fermentation, which comprises the steps of activating seeds of escherichia coli, bacillus subtilis and staphylococcus to prepare a mixed starter culture, inoculating the mixed starter culture into a fermentation medium containing glucose and a nitrogen source, carrying out constant-temperature shaking table culture to obtain a fermentation broth containing glucosamine, separating and purifying, and further preparing and purifying the glucosamine potassium sulfate. The fermentation activity of the escherichia coli is improved by the mixed starter, the production speed of the glutamic acid in the fermentation process can be reduced by adding the cysteine and the alanine, the synthesis rate of the glucosamine is increased, the purity of the product is improved, and the purity of the final product, namely the glucosamine potassium sulfate is improved.

Description

Method for producing glucosamine potassium sulfate salt by microbial fermentation
Technical Field
The invention belongs to the technical field of biological fermentation, and particularly relates to a method for producing glucosamine potassium sulfate salt by microbial fermentation.
Background
Glucosamine (GleN) is an important hexosamine formed by substituting one hydroxyl group of glucose with an amino group, and there are two main types of glucosamine on the market today, one is glucosamine hydrochloride and the other is glucosamine sulfate. D-Glucosamine Hydrochloride (D-Glucosamine Hydrochloride), molecular formula C6H13NO5HCl, a white crystal, odorless, slightly sweet, easily soluble in water, slightly soluble in methanol, insoluble in organic solvents such as ethanol, has important physiological functions for human body, participates in liver and kidney detoxification, plays a role in anti-inflammation and liver protection, has good curative effect on rheumatic arthritis and gastric ulcer, and is a main raw material for synthesizing antibiotics and anticancer drugs; can also be used in food, cosmetic and feed additive. Glucosamine hydrochloride is extracted from natural chitin, is a marine biological agent, and is the main component of chondroitin sulfate. It can promote the synthesis of mucopolysaccharide, raise the viscosity of joint synovial fluid, improve the metabolism of joint cartilage, promote the repair of joint cartilage and has obvious antiphlogistic and analgesic effects. It has the effect of promoting the injection efficiency of antibiotics, and can be used as nutritional supplement for diabetic patients.
The current methods for producing glucosamine 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 from exoskeletons of fishes, shrimps, crabs and the like, and glucosamine is obtained by extracting chitin and chitosan and then carrying out acidolysis or enzymolysis; however, a large amount of concentrated hydrochloric acid is needed in the acid hydrolysis process, which can cause serious industrial pollution; the enzymolysis method is to degrade the exoskeleton of fishes, shrimps and crabs by using chitosan, and has low process efficiency and higher production cost, so that the search for a proper microbial fermentation method to realize the industrial production of glucosamine is the current environmental and market demand. Escherichia coli is a common glucosamine synthetase producing strain. The research shows that: in Escherichia coli, glucosamine is produced from glutamine as an amino donor and fructose-6-phosphate under the catalytic action of glucosamine synthetase (GlmS). However, in the prior art, the production amount of glucosamine is low, the industrial high-efficiency production is difficult to realize, and the product purity can not reach the eyeball of a consumer.
Disclosure of Invention
In order to make up the defects of the prior art, the invention provides the method for producing glucosamine potassium sulfate by microbial fermentation, which has the advantages of high production efficiency, low cost and simple operation.
The invention is realized by the following technical scheme:
a method for producing glucosamine potassium sulfate by microbial fermentation is characterized in that: the method comprises the following steps:
(1) activating the seeds of escherichia coli, bacillus subtilis and staphylococcus to prepare a mixed starter;
(2) inoculating the mixed leaven into a fermentation medium containing glucose and a nitrogen source for constant temperature shaking table culture, controlling the rotation speed of the shaking table to be 230-;
(3) the fermentation process is carried out in a constant-speed feeding manner, the glucose feeding speed is controlled to be 3-5L/h, the pH value in the fermentation process is controlled to be 6.8-7.0 by ammonia water, and the fermentation culture is carried out for 35-48h to obtain fermentation liquor;
(4) centrifuging the fermentation liquor, removing thallus, collecting supernatant, adding hydrochloric acid into the supernatant, and hydrolyzing to obtain hydrolysate;
(5) concentrating the hydrolysate in vacuum at 75-90 deg.c and vacuum degree of-0.05-0.1 MPa, and recovering hydrochloric acid;
(6) cooling the concentrated solution to 5-10 ℃, crystallizing for 5-7h under stirring, precipitating for 3-4h, centrifuging to obtain glucosamine hydrochloride crude product, and adding concentrated hydrochloric acid into the mother solution obtained by centrifuging for hydrolyzing the concentrated solution of airplane after staying up;
(7) dissolving the obtained crude product with water, subjecting the water solution to pretreated macroporous adsorbent resin column chromatography, eluting with water until no chloride ion exists, concentrating under reduced pressure, and recrystallizing to obtain glucosamine hydrochloride;
(8) dissolving glucosamine hydrochloride in water, adding potassium sulfate under stirring, stirring at 40-50 deg.C for 30-60min, and freeze drying the reaction product under vacuum to obtain glucosamine potassium sulfate.
Preferably, the Escherichia coli, the Bacillus subtilis and the staphylococcus in the step (1) are activated and cultured on a plate culture medium for 6-8h before being subjected to seed activation.
Further, the plate culture medium in the step (1) comprises 11.5g/L of fish peptone, 6g/L of yeast extract, 5g/L of sodium chloride, 2g/L of ammonium sulfate and 15g/L of agar.
Further, the mixed starter in the step (1) is prepared according to the volume ratio of 3 (1-2) to 0.8-1 of escherichia coli, bacillus subtilis and staphylococcus.
Further, the seeds of the escherichia coli, the bacillus subtilis and the staphylococcus are activated in the step (1) by inoculating strains in a seed culture medium, controlling the culture temperature to be 36-38 ℃, the shaking table speed to be 220-270rpm, and the culture time to be 12-14 h.
Further, the seed culture medium in the step (1) comprises 11.5g/L of fish peptone, 20g/L of yeast extract powder, 5g/L of sodium chloride, 2g/L of ammonium sulfate, 5.5g/L of glycerol and 6.8-7.0 of pH.
Preferably, the fermentation medium in step (2) comprises: 25g/L glucose, 20g/L yeast extract powder, 10g/L alanine, 10g/L cysteine, 3g/L sodium nitrate, 1.05g/L dipotassium phosphate, 0.45g/L potassium dihydrogen phosphate, 0.1g/L sodium chloride, 0.5g/L magnesium sulfate, 0.03g/L ferrous sulfate, 3.2g/L lactose and 5.5g/L glycerol.
Further, the mixed starter in the step (2) is inoculated on the fermentation medium according to the inoculation amount of 10%.
Preferably, the hydrochloric acid in the step (4) is 30% concentrated hydrochloric acid.
Preferably, the stirring speed of the stirring crystallization in the step (6) is 50 rpm.
Preferably, the macroporous adsorption resin column in the step (7) is 732 cation exchange columns; the flow rate of the aqueous solution passing through the column bed is 3-5ml cm-2·min-1(ii) a The eluted solution is 3-5% AgNO3The solution checks the eluate for chloride ions.
Further, in the step (7), recrystallization is carried out by using an 85-95% ethanol solution.
Preferably, the mass ratio of the potassium sulfate, the glucosamine hydrochloride and the water in the step (8) is 1 (2-4) to (10-15).
The invention has the beneficial effects that: according to the invention, the fermentation activity of escherichia coli is improved by the mixed leaven, the glucosamine is efficiently produced by fermentation, the production speed of glutamic acid in the fermentation process can be reduced by adding cysteine and alanine, the synthesis rate of the glucosamine is improved, the purity of the product is improved by acidolysis, concentration, crystallization and decoloration, the reaction cost is reduced, and the yield of the product is improved.
Detailed Description
The present invention will be described in further detail with reference to specific embodiments thereof to assist those skilled in the art in providing a more complete, accurate and thorough understanding of the inventive concept and aspects thereof, and the scope of the present invention includes, but is not limited to, the following examples, and any modifications in the details and form of the technical aspects thereof that fall within the spirit and scope of the present application are intended to be included therein.
The following examples all follow the following medium composition:
the plate culture medium comprises: 11.5g/L of fish peptone, 6g/L of yeast extract powder, 5g/L of sodium chloride, 2g/L of ammonium sulfate and 15g/L of agar.
The seed culture medium comprises: 11.5g/L of fish peptone, 20g/L of yeast extract powder, 5g/L of sodium chloride, 2g/L of ammonium sulfate and 5.5g/L of glycerol;
the fermentation medium comprises: 25g/L glucose, 20g/L yeast extract powder, 10g/L alanine, 10g/L cysteine, 3g/L sodium nitrate, 1.05g/L dipotassium phosphate, 0.45g/L potassium dihydrogen phosphate, 0.1g/L sodium chloride, 0.5g/L magnesium sulfate, 0.03g/L ferrous sulfate, 3.2g/L lactose and 5.5g/L glycerol.
Example 1
Activating and culturing Escherichia coli, Bacillus subtilis and Staphylococcus on a plate culture medium for 6h, controlling the culture temperature at 37 deg.C and shaking table speed at 200 rpm; mixing cultured Escherichia coli, Saccharomyces cerevisiae and Staphylococcus aureus at volume ratio of 3:1:0.8, inoculating into seed culture medium, activating, culturing at 37 deg.C at shaking table speed of 230rpm for 12h, and adjusting pH to 6.9 with ammonia water; inoculating the mixed starter to a fermentation medium containing glucose and a nitrogen source according to the inoculation amount of 10%, carrying out fermentation culture in a fermentation tank at the culture temperature of 37 ℃, the shaking table speed of 230rpm for 40h, adjusting the pH value to 6.9 by using ammonia water to obtain fermentation liquor containing glucosamine, and controlling the glucose feeding speed to be 4L/h in the whole fermentation process; centrifuging the fermentation liquor, removing thallus, collecting supernatant, adding 30% hydrochloric acid into the supernatant, and hydrolyzing to obtain hydrolysate; concentrating the hydrolysate at 80 deg.C under vacuum degree of-0.07 MPa, and recovering hydrochloric acid; cooling the concentrated solution to 10 ℃, crystallizing for 6 hours under the stirring of 50rpm, precipitating for 4 hours, carrying out centrifugal separation to obtain a glucosamine hydrochloride crude product, and adding concentrated hydrochloric acid into a mother solution obtained by centrifugation for hydrolyzing the concentrated solution when the airplane stays up; dissolving the crude product with water, subjecting the aqueous solution to pretreated 732 cation exchange column chromatography, and allowing the aqueous solution to pass through the column bed at flow rate of 3ml cm-2·min-1Eluting with water and 5% AgNO3Detecting the solution until no chloride ion exists, concentrating under reduced pressure, and recrystallizing with 95% ethanol solution to obtain glucosamine hydrochloride; the used resin bed can be regenerated by eluting with 95% ethanol; dissolving glucosamine hydrochloride in water, adding potassium sulfate under stirring, wherein the mass ratio of potassium sulfate to glucosamine hydrochloride to water is 1:2:10, stirring at 40 ℃ for 60min, and freeze-drying the reaction product at-35 ℃ under vacuum to obtain glucosamine potassium sulfate with the purity of 98.3%.
Example 2
Mixing Escherichia coli and Bacillus subtilis budActivating and culturing bacillus and staphylococcus on a plate culture medium for 6 hours, controlling the culture temperature to be 37 ℃ and the shaking table speed to be 200 rpm; mixing cultured Escherichia coli, Saccharomyces cerevisiae and Staphylococcus aureus at volume ratio of 3:1.2:0.8, inoculating into seed culture medium, activating, culturing at 36 deg.C and shaking table speed of 240rpm for 13h, and adjusting pH to 6.8 with ammonia water; inoculating the mixed starter culture into a fermentation medium containing glucose and a nitrogen source according to the inoculation amount of 10 percent, carrying out fermentation culture in a fermentation tank at the culture temperature of 36 ℃, the shaking table speed of 240rpm and the culture time of 38h, adjusting the pH value to 6.8 by using ammonia water to obtain fermentation liquor containing glucosamine, controlling the glucose feeding speed to be 4.5L/h in the whole fermentation process, carrying out centrifugal separation on the fermentation liquor, removing thalli, collecting supernatant, and adding 30 percent hydrochloric acid into the supernatant for hydrolysis to obtain hydrolysate; concentrating the hydrolysate at 80 deg.C under vacuum degree of-0.05 MPa, and recovering hydrochloric acid; cooling the concentrated solution to 5 ℃, crystallizing for 7 hours under the stirring of 50rpm, precipitating for 3 hours, carrying out centrifugal separation to obtain a glucosamine hydrochloride crude product, and adding concentrated hydrochloric acid into a mother solution obtained by centrifugation for hydrolyzing the concentrated solution when the airplane stays up; dissolving the crude product with water, subjecting the aqueous solution to pretreated 732 cation exchange column chromatography, and allowing the aqueous solution to pass through the column bed at flow rate of 3ml cm-2·min-1Eluting with water and 5% AgNO3Detecting the solution until no chloride ion exists, concentrating under reduced pressure, and recrystallizing with 95% ethanol solution to obtain glucosamine hydrochloride; the used resin bed can be regenerated by eluting with 95% ethanol; dissolving glucosamine hydrochloride in water, adding potassium sulfate under stirring, wherein the mass ratio of potassium sulfate to glucosamine hydrochloride to water is 1:2:10, stirring at 40 ℃ for 60min, and freeze-drying the reaction product at-35 ℃ under vacuum to obtain glucosamine potassium sulfate with the purity of 97.8%.
Example 3
Activating and culturing Escherichia coli, Bacillus subtilis and Staphylococcus on a plate culture medium for 6h, controlling the culture temperature at 37 deg.C and shaking table speed at 200 rpm; mixing cultured Escherichia coli, Saccharomyces cerevisiae and Staphylococcus according to volume ratio of 3:1.5:0.7, inoculating into seed culture medium, activating and culturing at 38 deg.C and shaking table speed of 250rpm, culture time of 12h, and adjusting the pH value to 6.9 by using ammonia water; inoculating the mixed starter culture into a fermentation medium containing glucose and a nitrogen source according to the inoculation amount of 10 percent, carrying out fermentation culture in a fermentation tank at the culture temperature of 38 ℃, the shaking table speed of 250rpm for 46h, adjusting the pH value to 6.9 by using ammonia water to obtain fermentation liquor containing glucosamine, controlling the glucose feeding speed to be 4L/h in the whole fermentation process, carrying out centrifugal separation on the fermentation liquor, removing thalli, collecting supernatant, and adding 30 percent hydrochloric acid into the supernatant for hydrolysis to obtain hydrolysate; concentrating the hydrolysate at 75-90 deg.C under vacuum degree of-0.1 MPa, and recovering hydrochloric acid; cooling the concentrated solution to 10 ℃, crystallizing for 5 hours under the stirring of 50rpm, precipitating for 3 hours, carrying out centrifugal separation to obtain a glucosamine hydrochloride crude product, and adding concentrated hydrochloric acid into a mother solution obtained by centrifugation for hydrolyzing the concentrated solution when the airplane stays up; dissolving the crude product with water, subjecting the aqueous solution to pretreated 732 cation exchange column chromatography, and allowing the aqueous solution to pass through the column bed at flow rate of 3ml cm-2·min-1Eluting with water and 5% AgNO3Detecting the solution until no chloride ion exists, concentrating under reduced pressure, and recrystallizing with 95% ethanol solution to obtain glucosamine hydrochloride; the used resin bed can be regenerated by eluting with 95% ethanol; dissolving glucosamine hydrochloride in water, adding potassium sulfate under stirring, wherein the mass ratio of potassium sulfate to glucosamine hydrochloride to water is 1:2:10, stirring at 40 ℃ for 60min, and freeze-drying the reaction product at-35 ℃ under vacuum to obtain glucosamine potassium sulfate with the purity of 97.6%.
Example 4
Activating and culturing Escherichia coli, Bacillus subtilis and Staphylococcus on a plate culture medium for 6h, controlling the culture temperature at 37 deg.C and shaking table speed at 200 rpm; mixing cultured Escherichia coli, Saccharomyces cerevisiae and Staphylococcus aureus at volume ratio of 3:1.2:0.8, inoculating into seed culture medium, activating, culturing at 37 deg.C and shaking table speed of 230rpm for 14h, and adjusting pH to 7.0 with ammonia water; inoculating the mixed starter to a fermentation medium containing glucose and nitrogen source according to the inoculation amount of 10%, performing fermentation culture in a fermentation tank at 37 deg.C with shaking table speed of 230rpm for 48h, and adjusting pH to 6.9 with ammonia water to obtain the final productThe fermentation liquor containing glucosamine is subjected to centrifugal separation to remove thalli, supernatant is collected, and 30% hydrochloric acid is added into the supernatant for hydrolysis to obtain hydrolysate; concentrating the hydrolysate at 90 deg.C under vacuum degree of-0.1 MPa, and recovering hydrochloric acid; cooling the concentrated solution to 6 ℃, crystallizing for 6 hours under the stirring of 50rpm, precipitating for 4 hours, carrying out centrifugal separation to obtain a glucosamine hydrochloride crude product, and adding concentrated hydrochloric acid into a mother solution obtained by centrifugation for hydrolyzing the concentrated solution when the airplane stays up; dissolving the crude product with water, subjecting the aqueous solution to pretreated 732 cation exchange column chromatography, and allowing the aqueous solution to pass through the column bed at flow rate of 3ml cm-2·min-1Eluting with water and 5% AgNO3Detecting the solution until no chloride ion exists, concentrating under reduced pressure, and recrystallizing with 95% ethanol solution to obtain glucosamine hydrochloride; the used resin bed can be regenerated by eluting with 95% ethanol; dissolving glucosamine hydrochloride in water, adding potassium sulfate under stirring, wherein the mass ratio of potassium sulfate to glucosamine hydrochloride to water is 1:2:10, stirring at 40 ℃ for 60min, and freeze-drying the reaction product at-35 ℃ under vacuum to obtain glucosamine potassium sulfate with the purity of 97.9%.
Example 5
Activating and culturing Escherichia coli, Bacillus subtilis and Staphylococcus on a plate culture medium for 6h, controlling the culture temperature at 37 deg.C and shaking table speed at 200 rpm; mixing cultured Escherichia coli, Saccharomyces cerevisiae and Staphylococcus aureus at volume ratio of 3:2:0.5, inoculating into seed culture medium, activating, culturing at 36 deg.C under shaking table speed of 240rpm for 12h, and adjusting pH to 6.8 with ammonia water; inoculating the mixed starter culture into a fermentation medium containing glucose and a nitrogen source according to the inoculation amount of 10 percent, carrying out fermentation culture in a fermentation tank at the culture temperature of 36 ℃, the shaking table speed of 240rpm and the culture time of 35h, adjusting the pH value to 6.8 by using ammonia water to obtain fermentation liquor containing glucosamine, controlling the glucose feeding speed to be 3.5L/h in the whole fermentation process, carrying out centrifugal separation on the fermentation liquor, removing thalli, collecting supernatant, and adding 30 percent hydrochloric acid into the supernatant for hydrolysis to obtain hydrolysate; concentrating the hydrolysate at 90 deg.C under vacuum degree of-0.1 MPa, and recovering hydrochloric acid; will be concentratedCooling the concentrated solution to 10 ℃, crystallizing for 5 hours under the stirring of 50rpm, precipitating for 4 hours, carrying out centrifugal separation to obtain a glucosamine hydrochloride crude product, and adding concentrated hydrochloric acid into the mother solution obtained by centrifugation for hydrolyzing the concentrated solution after the airplane stays up; dissolving the crude product with water, subjecting the aqueous solution to pretreated 732 cation exchange column chromatography, and allowing the aqueous solution to pass through the column bed at flow rate of 3ml cm-2·min-1Eluting with water and 5% AgNO3Detecting the solution until no chloride ion exists, concentrating under reduced pressure, and recrystallizing with 95% ethanol solution to obtain glucosamine hydrochloride; the used resin bed can be regenerated by eluting with 95% ethanol; dissolving glucosamine hydrochloride in water, adding potassium sulfate under stirring, wherein the mass ratio of potassium sulfate to glucosamine hydrochloride to water is 1:2:10, stirring at 40 ℃ for 60min, and freeze-drying the reaction product at-35 ℃ under vacuum to obtain glucosamine potassium sulfate with the purity of 98.1%.

Claims (10)

1. A method for producing glucosamine potassium sulfate salt by microbial fermentation is characterized by comprising the following steps: the method comprises the following steps:
(1) activating the seeds of escherichia coli, bacillus subtilis and staphylococcus to prepare a mixed starter;
(2) inoculating the mixed leaven into a fermentation medium containing glucose and a nitrogen source for constant temperature shaking table culture, controlling the rotation speed of the shaking table to be 230-;
(3) the fermentation process is carried out in a constant-speed feeding manner, the glucose feeding speed is controlled to be 3-5L/h, the pH value in the fermentation process is controlled to be 6.8-7.0 by ammonia water, and the fermentation culture is carried out for 35-48h to obtain fermentation liquor;
(4) centrifuging the fermentation liquor, removing thallus, collecting supernatant, adding hydrochloric acid into the supernatant, and hydrolyzing to obtain hydrolysate;
(5) concentrating the hydrolysate in vacuum at 75-90 deg.c and vacuum degree of-0.05-0.1 MPa;
(6) cooling the concentrated solution to 5-10 deg.C, crystallizing under stirring for 5-7 hr, precipitating for 3-4 hr, and centrifuging to obtain glucosamine hydrochloride crude product;
(7) dissolving the obtained crude product with water, subjecting the water solution to pretreated macroporous adsorbent resin column chromatography, eluting with water until no chloride ion exists, concentrating under reduced pressure, and recrystallizing to obtain glucosamine hydrochloride;
(8) dissolving glucosamine hydrochloride in water, adding potassium sulfate under stirring, stirring at 40-50 deg.C for 30-60min, and freeze drying the reaction product under vacuum to obtain glucosamine potassium sulfate.
2. The method for producing glucosamine potassium sulfate by microbial fermentation according to claim 1, wherein the method comprises the following steps: and (2) performing activation culture on the escherichia coli, the bacillus subtilis and the staphylococcus for 6-8h on a plate culture medium before performing seed activation.
3. The method for producing glucosamine potassium sulfate by microbial fermentation according to claim 2, wherein the method comprises the following steps: the plate culture medium in the step (1) comprises 11.5g/L of fish peptone, 6g/L of yeast extract powder, 5g/L of sodium chloride, 2g/L of ammonium sulfate and 15g/L of agar.
4. The method for producing glucosamine potassium sulfate by microbial fermentation according to claim 1 or 2, wherein: the mixed leaven in the step (1) is prepared according to the volume ratio of 3 (1-2) to 0.8-1) of escherichia coli, bacillus subtilis and staphylococcus.
5. The method for producing glucosamine potassium sulfate by microbial fermentation according to claim 1 or 2, wherein: the seeds of the escherichia coli, the bacillus subtilis and the staphylococcus in the step (1) are activated by inoculating strains in a seed culture medium, controlling the culture temperature to be 36-38 ℃, the shaking table speed to be 220-270rpm, and the culture time to be 12-14 h.
6. The method for producing glucosamine potassium sulfate by microbial fermentation according to claim 5, wherein the method comprises the following steps: the seed culture medium in the step (1) comprises 11.5g/L of fish peptone, 20g/L of yeast extract powder, 5g/L of sodium chloride, 2g/L of ammonium sulfate, 5.5g/L of glycerol and pH 6.8-7.0.
7. The method for producing glucosamine potassium sulfate by microbial fermentation according to claim 1, wherein the method comprises the following steps: the fermentation medium in the step (2) comprises: 25g/L glucose, 20g/L yeast extract powder, 10g/L alanine, 10g/L cysteine, 3g/L sodium nitrate, 1.05g/L dipotassium phosphate, 0.45g/L potassium dihydrogen phosphate, 0.1g/L sodium chloride, 0.5g/L magnesium sulfate, 0.03g/L ferrous sulfate, 3.2g/L lactose and 5.5g/L glycerol.
8. The method for producing glucosamine potassium sulfate by microbial fermentation according to claim 1, wherein the method comprises the following steps: the macroporous adsorption resin column in the step (7) is a 732 cation exchange column; the flow rate of the aqueous solution passing through the column bed is 3-5ml cm-2·min-1(ii) a The eluted solution is 3-5% AgNO3The solution checks the eluate for chloride ions.
9. The method for producing glucosamine potassium sulfate by microbial fermentation according to claim 1, wherein the method comprises the following steps: and (4) recrystallizing in 85-95% ethanol solution in the step (7).
10. The method for producing glucosamine potassium sulfate by microbial fermentation according to claim 1, wherein the method comprises the following steps: in the step (8), the mass ratio of the potassium sulfate to the glucosamine hydrochloride to the water is 1 (2-4) to 10-15.
CN201911210934.7A 2019-12-02 2019-12-02 Method for producing glucosamine potassium sulfate salt by microbial fermentation Withdrawn CN110846373A (en)

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CN113045610A (en) * 2020-12-20 2021-06-29 宁夏金维制药股份有限公司 Method for extracting glucosamine from N-acetylglucosamine fermentation liquor
CN114853826A (en) * 2022-05-12 2022-08-05 山西辅仁恒峰药业有限公司 Preparation method of glucosamine sulfate sodium chloride double salt

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113045610A (en) * 2020-12-20 2021-06-29 宁夏金维制药股份有限公司 Method for extracting glucosamine from N-acetylglucosamine fermentation liquor
CN114853826A (en) * 2022-05-12 2022-08-05 山西辅仁恒峰药业有限公司 Preparation method of glucosamine sulfate sodium chloride double salt

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