CN112125935A - Preparation method of rhamnose - Google Patents
Preparation method of rhamnose Download PDFInfo
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- CN112125935A CN112125935A CN202011101481.7A CN202011101481A CN112125935A CN 112125935 A CN112125935 A CN 112125935A CN 202011101481 A CN202011101481 A CN 202011101481A CN 112125935 A CN112125935 A CN 112125935A
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Abstract
The invention relates to the technical field of separation and purification of rhamnose, and particularly relates to a preparation method of rhamnose. When sugar liquid containing rhamnose and glucose is treated, glucose oxidase and catalase are jointly used, glucose in the sugar liquid can be effectively removed, rhamnose with the purity of more than 98% is separated, and the yield of the rhamnose is more than 95%. In addition, through the method, the rhamnose is separated and purified, the filtrate is decolorized and then directly concentrated and crystallized in the water phase to obtain the white and clean rhamnose, an organic solvent is not required to be used, the high production cost caused by the use of a large amount of organic solvents and the influence on the environment caused by the recovery and treatment of the organic solvents are avoided, and the method is green and environment-friendly, simple in process and high in purification efficiency.
Description
Technical Field
The invention relates to the technical field of separation and purification of rhamnose, and particularly relates to a preparation method of rhamnose.
Background
Rhamnose is also called mannose, is methyl pentose, has 2 isomers of L-type and D-type, pure rhamnose is colorless crystalline powder, can be dissolved in water and methanol, is slightly soluble in ethanol, and has two forms of crystallization, namely alpha form and beta form, wherein the alpha form contains a molecule of crystal water, the crystal water is lost after heating, the beta form is converted into the beta form, the beta form is easy to absorb moisture, and the alpha form is converted into the alpha form by absorbing moisture in the air, and the alpha-L-rhamnose is common.
The conventional method for producing rhamnose is to extract from natural plants, such as quercitrin from oak bark, naringin from orange peel or vitamin A from bark of Tochis. The rhamnose contained in the molecules of the substances can be hydrolyzed to obtain rhamnose, and then the rhamnose is extracted and subjected to other chemical unit process operations to obtain a pure rhamnose product. However, this method has major disadvantages, such as high labor intensity, generation of a large amount of potential toxic substances during the extraction process, use of toxic or corrosive chemical reagents during the extraction process, and inconvenience in production due to seasonal and regional sources of raw materials and transportation.
The method for industrially producing rhamnose is mainly a fermentation method. The method for producing rhamnose can be divided into two types according to the difference of intermediate products, (1) polysaccharide containing rhamnosyl is produced by using microorganisms or algae, and the polysaccharide is hydrolyzed to obtain rhamnose; (2) the rhamnolipid is produced by microorganisms, and the rhamnolipid is hydrolyzed to obtain the rhamnose. The microorganisms commonly selected for producing rhamnose by a fermentation method comprise yeast, pseudomonas, klebsiella, acinetobacter and lactobacillus, wherein pseudomonas aeruginosa in pseudomonas is the most commonly used. The carbon source in the culture medium is mostly selected from oily substrates which can induce the production of rhamnolipid. The common method comprises the following steps: the specific degradation capability of yeast to glucose is utilized to degrade glucose first and then crystallize rhamnose. The optimal preparation process is that the temperature is 30 ℃, the pH value is 5.0, and the yeast amount is 2 percent of the mother liquor.
The waste liquid generated by the plant extraction process of some active substances contains rhamnose and glucose, and for the recycling of the waste liquid, the problem that how to remove glucose impurities to improve the purity of rhamnose finished products is usually faced, the glucose is difficult to remove, and the recovery rate and the purity of the rhamnose are decisive indexes for measuring the feasibility of the waste liquid recycling process. The existing purification process usually needs a large amount of organic solvent, the recovery treatment of the organic solvent has great influence on the environment, the cost is high, the production period for separating rhamnose is long, and the purity of the rhamnose is difficult to reach more than 98%. Therefore, a large amount of waste liquid containing rhamnose and glucose is simply treated, and waste and pollution are caused.
Disclosure of Invention
The invention provides a preparation method of rhamnose, which does not need to use an organic solvent and has high purity and high yield of rhamnose, aiming at the problems of large consumption of the organic solvent and low purity or low yield of a rhamnose finished product in the existing technology for recovering rhamnose from a sugar solution containing rhamnose and glucose.
In order to achieve the purpose, the invention adopts the following technical scheme.
A preparation method of rhamnose comprises the following steps:
and S1, dissolving the sugar solution in water to obtain a reaction sugar solution.
The sugar solution is a mixed solution containing rhamnose and glucose; the total mass of rhamnose and glucose in the reaction sugar solution is less than or equal to 35%.
Preferably, the sum of the masses of rhamnose and glucose in the reaction sugar solution is 25-35% of the reaction sugar solution.
Preferably, the water is purified water subjected to sterilization treatment.
More preferably, the sterilization treatment is heating purified water to 100 ℃ and keeping the temperature for more than 30 min.
In the steps, after the water is heated to 100 ℃ and is kept warm for 30min, the water is naturally cooled to 50-55 ℃, and sugar solution is added into the water.
S2, adjusting the pH value of the reaction sugar liquid to 5.5-6.0 by using calcium hydroxide, then placing the reaction sugar liquid in an environment of 50-55 ℃, and continuously oxygenating the reaction sugar liquid.
Preferably, the pH of the reaction sugar solution is adjusted to 5.5 with calcium hydroxide.
Preferably, the reaction sugar solution is placed in an environment at 52 ℃.
More preferably, the reaction sugar solution is placed in a water bath kettle at 50-55 ℃.
S3, adding glucose oxidase and catalase into the reaction sugar solution to obtain reaction liquid, and reacting the reaction liquid for 40-48 hours at 50-55 ℃ in an oxygenated state; the pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5-6.0, and the mass of the reaction solution was 75-125% of the initial mass by supplementing water.
The addition amount of the glucose oxidase and the catalase is 2-3% of the mass of the reaction sugar solution.
Preferably, the addition amount of the glucose oxidase and the catalase is 2.5% of the mass of the reaction sugar solution.
And S4, placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, decoloring the filtrate with activated carbon, and concentrating and crystallizing to obtain rhamnose.
In the above steps S2 and S3, oxygen is introduced into the reaction sugar solution or the reaction solution in an amount of 1L/min or more per kg of the reaction sugar solution or the reaction solution.
Compared with the prior art, the invention has the beneficial effects that:
when sugar liquid containing rhamnose and glucose is treated, glucose oxidase and catalase are jointly used, glucose in the sugar liquid can be effectively removed, rhamnose with the purity of more than 98% is separated, and the yield of the rhamnose is more than 95%. In addition, through the method, the rhamnose is separated and purified, and after the filtrate is decolorized, the filtrate is directly concentrated and crystallized in a water phase to obtain the white and clean rhamnose, an organic solvent is not required, so that the high production cost caused by the use of a large amount of organic solvents and the influence on the environment caused by the recovery and treatment of the organic solvents are avoided.
The method can completely convert glucose in the sugar solution into calcium gluconate by controlling the dosage of enzyme, the content of sugar in the reaction solution and the temperature and pH value of the reaction solution in the reaction process, thereby obtaining high-purity rhamnose crystal in the subsequent crystallization step.
Drawings
FIG. 1 is a high performance liquid chromatogram of a finished rhamnose product prepared in example 1;
fig. 2 is a high performance liquid chromatogram of the finished rhamnose product prepared in example 2.
Detailed Description
In order to more fully understand the technical contents of the present invention, the technical solutions of the present invention will be further described and illustrated with reference to the following specific embodiments.
The sugar solution treated in the following examples is a mixed solution mainly containing rhamnose and glucose, and also contains a small amount of inorganic salt impurities and other residual plant extracts, wherein the rhamnose content in the sugar solution is 43% by mass, and the glucose content in the sugar solution is 36% by mass. In the method, the sugar solution is diluted by sterilized water to form a reaction sugar solution, and the sum of the mass percentages of glucose and rhamnose in the reaction sugar solution is controlled within 35 percent. The method for sterilizing purified water comprises heating water to 100 deg.C and maintaining the temperature for 30 min.
The purity of rhamnose separated from the sugar solution was analyzed by high performance liquid chromatography under the following chromatographic conditions. A chromatographic column: kromil amino column 4.6 μm × 250 mm; mobile phase: acetonitrile water 70:30 (filtered and ultrasonically degassed before use); flow rate: 1 mL/min; refractive index detector temperature: 30 ℃; sample introduction amount: 10 μ L.
Example 1
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. 358g of sugar solution is weighed, when purified water is naturally cooled to 52 ℃, the sugar solution is dissolved in the purified water and uniformly stirred, so that the sugar solution is completely dissolved to form a reaction sugar solution, the mass percentage of glucose in the reaction sugar solution is 16%, and the sum of the mass percentages of glucose and rhamnose is 35%.
(2) Adjusting pH of the reaction sugar solution to 5.5 with calcium hydroxide, placing the reaction sugar solution in a 52 deg.C water bath, and oxygenating with oxygen at 1L/min.
(3) Respectively adding 20mL of glucose oxidase (1.00g/mL, 20 ℃) and 20mL of catalase (1.06g/mL, 20 ℃) into the reaction sugar liquid to obtain reaction liquid, reacting the reaction liquid for 45 hours at 52 ℃ in an oxygenated state (oxygen introduction amount is 1L/min), and measuring the content of glucose in the reaction liquid to obtain the content of glucose in the reaction liquid, wherein the content of glucose in the reaction liquid is 0; the pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5, and the mass of the reaction solution was made the initial mass (about 845g) by supplementing sterilized purified water.
(4) And (3) placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate with activated carbon, concentrating, crystallizing and drying to obtain 164.5g of a white and clean rhamnose finished product, wherein the yield is 97.1%.
The purity of rhamnose separated in this example was analyzed by high performance liquid chromatography, and the high performance liquid chromatogram is shown in fig. 1, where the retention time of rhamnose is 8.250min, the peak area is 98.1807%, i.e., the purity is 98.1807%.
Example 2
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. Weighing 206g of sugar solution, dissolving the sugar solution in purified water when the purified water is naturally cooled to 52 ℃, uniformly stirring to completely dissolve the sugar solution to form a reaction sugar solution, wherein the mass percentage of glucose in the reaction sugar solution is 11.4%, and the sum of the mass percentages of glucose and rhamnose is 25%.
(2) Adjusting pH of the reaction sugar solution to 5.5 with calcium hydroxide, placing the reaction sugar solution in a 52 deg.C water bath, and oxygenating with oxygen at 1L/min.
(3) Respectively adding 16mL of glucose oxidase (1.00g/mL, 20 ℃) and 16mL of catalase (1.06g/mL, 20 ℃) into the reaction sugar liquid to obtain reaction liquid, reacting the reaction liquid for 45 hours at 52 ℃ in an oxygenated state (oxygen introduction amount is 1L/min), and measuring the content of glucose in the reaction liquid to obtain the content of glucose in the reaction liquid, wherein the content of glucose in the reaction liquid is 0; the pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5, and the mass of the reaction solution was made the initial mass (about 684g) by supplementing sterilized purified water.
(4) And (3) placing the reaction solution at room temperature, neutralizing the reaction solution by using calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate by using activated carbon, concentrating, crystallizing and drying to obtain 95.3g of a white rhamnose finished product, wherein the yield is 96.8%.
The purity of rhamnose separated in this example was analyzed by high performance liquid chromatography, and the high performance liquid chromatogram is shown in fig. 2, where the retention time of rhamnose is 9.356min, and the peak area is 99.3997%, that is, the purity is 99.3997%.
Example 3
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. Weighing 304g of sugar solution, dissolving the sugar solution in purified water when the purified water is naturally cooled to 52 ℃, uniformly stirring to completely dissolve the sugar solution to form a reaction sugar solution, wherein the mass percentage of glucose in the reaction sugar solution is 14.6%, and the sum of the mass percentages of glucose and rhamnose is 32%.
(2) Adjusting pH of the reaction sugar solution to 5.5 with calcium hydroxide, placing the reaction sugar solution in a 52 deg.C water bath, and oxygenating with oxygen at 1L/min.
(3) Adding 19mL of glucose oxidase (1.00g/mL, 20 ℃) and 19mL of catalase (1.06g/mL, 20 ℃) into the reaction sugar solution respectively to obtain reaction liquid, reacting the reaction liquid for 45 hours at 52 ℃ in an oxygenated state (oxygen introduction amount is 1L/min), and measuring the content of glucose in the reaction liquid to obtain the content of glucose in the reaction liquid, wherein the content of glucose in the reaction liquid is 0; the pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5, and the mass of the reaction solution was made the initial mass (about 790g) by supplementing sterilized purified water.
(4) And (3) placing the reaction solution at room temperature, neutralizing the reaction solution by using calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate by using activated carbon, concentrating, crystallizing and drying to obtain 138.6g of a white and clean rhamnose finished product, wherein the yield is 95.6%.
The purity of the rhamnose separated in this example was analyzed by high performance liquid chromatography, the retention time of rhamnose was 9.013min, the peak area was 98.4599%, i.e. the purity was 98.4599%.
Example 4
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. 358g of sugar solution is weighed, when purified water is naturally cooled to 52 ℃, the sugar solution is dissolved in the purified water and uniformly stirred, so that the sugar solution is completely dissolved to form a reaction sugar solution, the mass percentage of glucose in the reaction sugar solution is 16%, and the sum of the mass percentages of glucose and rhamnose is 35%.
(2) Adjusting pH of the reaction sugar solution to 5.5 with calcium hydroxide, placing the reaction sugar solution in a 52 deg.C water bath, and oxygenating with oxygen at 1L/min.
(3) Respectively adding 16mL of glucose oxidase (1.00g/mL, 20 ℃) and 16mL of catalase (1.06g/mL, 20 ℃) into the reaction sugar liquid to obtain reaction liquid, reacting the reaction liquid for 45 hours at 52 ℃ in an oxygenated state (oxygen introduction amount is 1L/min), and measuring the content of glucose in the reaction liquid to obtain the content of glucose in the reaction liquid, wherein the content of glucose in the reaction liquid is 0; the pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5, and the mass of the reaction solution was made the initial mass (about 836g) by supplementing sterilized purified water.
(4) And (3) placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate with active carbon, concentrating, crystallizing and drying to obtain 162.3g of a white and clean rhamnose finished product, wherein the yield is 95.8%.
The purity of the rhamnose separated in this example was analyzed by high performance liquid chromatography, the retention time of rhamnose was 8.716min, the peak area was 98.2039%, i.e. the purity was 98.2039%.
Example 5
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. 358g of sugar solution is weighed, when purified water is naturally cooled to 52 ℃, the sugar solution is dissolved in the purified water and uniformly stirred, so that the sugar solution is completely dissolved to form a reaction sugar solution, the mass percentage of glucose in the reaction sugar solution is 16%, and the sum of the mass percentages of glucose and rhamnose is 35%.
(2) Adjusting pH of the reaction sugar solution to 5.5 with calcium hydroxide, placing the reaction sugar solution in a 52 deg.C water bath, and oxygenating with oxygen at 1L/min.
(3) Respectively adding 24mL of glucose oxidase (1.00g/mL, 20 ℃) and 24mL of catalase (1.06g/mL, 20 ℃) into the reaction sugar liquid to obtain reaction liquid, reacting the reaction liquid for 45 hours at 52 ℃ in an oxygenated state (oxygen introduction amount is 1L/min), and measuring the content of glucose in the reaction liquid to obtain the content of glucose in the reaction liquid, wherein the content of glucose in the reaction liquid is 0; the pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5, and the mass of the reaction solution was made the initial mass (about 852g) by supplementing sterilized purified water.
(4) And (3) placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate with active carbon, concentrating, crystallizing and drying to obtain 163.9g of white rhamnose finished product, wherein the yield is 96.7%.
The purity of the rhamnose separated in this example was analyzed by high performance liquid chromatography, the retention time of rhamnose was 8.685min, the peak area was 98.5031%, i.e. the purity was 98.5031%.
Example 6
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. 358g of sugar solution is weighed, when purified water is naturally cooled to 55 ℃, the sugar solution is dissolved in the purified water and uniformly stirred, so that the sugar solution is completely dissolved to form a reaction sugar solution, the mass percentage of glucose in the reaction sugar solution is 16%, and the sum of the mass percentages of glucose and rhamnose is 35%.
(2) Adjusting pH of the reacted sugar solution to 5.5 with calcium hydroxide, placing the reacted sugar solution in a water bath kettle at 55 deg.C, and oxygenating the reacted sugar solution with oxygen flow of 1L/min.
(3) Adding 20mL of glucose oxidase (1.00g/mL, 20 ℃) and 20mL of catalase (1.06g/mL, 20 ℃) into the reaction sugar liquid respectively to obtain reaction liquid, and reacting the reaction liquid for 45 hours at 55 ℃ in an oxygenated state (oxygen introduction amount is 1L/min); the pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5, and the mass of the reaction solution was made the initial mass (about 844g) by supplementing sterilized purified water.
(4) And (3) placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate with active carbon, concentrating, crystallizing and drying to obtain 161.4g of a white and clean rhamnose finished product, wherein the yield is 95.3%.
The purity of the rhamnose separated in this example was analyzed by high performance liquid chromatography, the retention time of rhamnose was 8.8921min, the peak area was 98.0038%, i.e. the purity was 98.0038%.
Example 7
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. 358g of sugar solution is weighed, when purified water is naturally cooled to 52 ℃, the sugar solution is dissolved in the purified water and uniformly stirred, so that the sugar solution is completely dissolved to form a reaction sugar solution, the mass percentage of glucose in the reaction sugar solution is 16%, and the sum of the mass percentages of glucose and rhamnose is 35%.
(2) Adjusting pH of the reaction sugar solution to 6.0 with calcium hydroxide, placing the reaction sugar solution in a 52 deg.C water bath, and oxygenating with oxygen at 1L/min.
(3) Adding 20mL of glucose oxidase (1.00g/mL, 20 ℃) and 20mL of catalase (1.06g/mL, 20 ℃) into the reaction sugar liquid respectively to obtain reaction liquid, and reacting the reaction liquid for 45 hours at 52 ℃ in an oxygenated state (oxygen introduction amount is 1L/min); the pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 6.0, and the mass of the reaction solution was made the initial mass (about 844g) by supplementing sterilized purified water.
(4) And (3) placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate with active carbon, concentrating, crystallizing and drying to obtain 161.0g of a white and clean rhamnose finished product, wherein the yield is 95.0%.
The purity of the rhamnose separated in this example was analyzed by high performance liquid chromatography, the retention time of rhamnose was 8.7469min, the peak area was 98.2635%, i.e. the purity was 98.2635%.
Example 8
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. 358g of sugar solution is weighed, when purified water is naturally cooled to 52 ℃, the sugar solution is dissolved in the purified water and uniformly stirred, so that the sugar solution is completely dissolved to form a reaction sugar solution, the mass percentage of glucose in the reaction sugar solution is 16%, and the sum of the mass percentages of glucose and rhamnose is 35%.
(2) Adjusting pH of the reaction sugar solution to 5.5 with calcium hydroxide, placing the reaction sugar solution in a 52 deg.C water bath, and oxygenating with oxygen at 1L/min.
(3) Adding 20mL of glucose oxidase (1.00g/mL, 20 ℃) into the reaction sugar solution to obtain a reaction solution, reacting the reaction solution for 45 hours at 52 ℃ in an oxygenated state (oxygen introduction amount is 1L/min), and measuring the content of glucose in the reaction solution, wherein the content of glucose is 9.6%; the reaction solution is continuously reacted for 15 hours under the same condition, and then the content of glucose in the solution is measured, wherein the content of glucose is 9.1 percent; the reaction was then terminated. The pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5, and the mass of the reaction solution was made the initial mass (about 824g) by supplementing sterilized purified water.
(4) And (3) placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate with active carbon, concentrating, crystallizing and drying to obtain 240.1g of powder, wherein the yield is 143.2%, and the powder contains a large amount of glucose.
Example 9
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. 358g of sugar solution is weighed, when purified water is naturally cooled to 52 ℃, the sugar solution is dissolved in the purified water and uniformly stirred, so that the sugar solution is completely dissolved to form a reaction sugar solution, the mass percentage of glucose in the reaction sugar solution is 16%, and the sum of the mass percentages of glucose and rhamnose is 35%.
(2) Adjusting pH of the reaction sugar solution to 5.5 with calcium hydroxide, placing the reaction sugar solution in a 52 deg.C water bath, and oxygenating with oxygen at 1L/min.
(3) Adding 40mL of glucose oxidase (1.00g/mL, 20 ℃) into the reaction sugar solution to obtain a reaction solution, reacting the reaction solution for 45 hours at 52 ℃ in an oxygenated state (oxygen introduction amount is 1L/min), and measuring the content of glucose in the reaction solution, wherein the content of glucose is 7.9%; the reaction solution is continuously reacted for 15 hours under the same condition, and then the content of glucose in the solution is measured, wherein the content of glucose is 7.3 percent; the reaction was then terminated. The pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5, and the mass of the reaction solution was made the initial mass (about 844g) by supplementing sterilized purified water.
(4) And (3) placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate with activated carbon, concentrating, crystallizing and drying to obtain 219.8g of powder, wherein the yield is 129.7%, and the powder contains a large amount of glucose.
Example 10
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. 358g of sugar solution is weighed, when purified water is naturally cooled to 52 ℃, the sugar solution is dissolved in the purified water and uniformly stirred, so that the sugar solution is completely dissolved to form a reaction sugar solution, the mass percentage of glucose in the reaction sugar solution is 16%, and the sum of the mass percentages of glucose and rhamnose is 35%.
(2) Adjusting pH of the reaction sugar solution to 5.5 with calcium hydroxide, placing the reaction sugar solution in a 52 deg.C water bath, and oxygenating with oxygen at 1L/min.
(3) Adding 10mL of glucose oxidase (1.00g/mL, 20 ℃) and 10mL of catalase (1.06g/mL, 20 ℃) into the reaction sugar liquid respectively to obtain reaction liquid, and reacting the reaction liquid for 48 hours at 52 ℃ in an oxygenated state (oxygen introduction amount is 1L/min); measuring the content of glucose in the reaction solution, wherein the content of glucose is 7.2%; the reaction solution is continuously reacted for 24 hours under the same condition, and the content of glucose in the solution is measured, wherein the content of glucose is 6.4 percent; then the reaction solution is continuously reacted for 24 hours under the same condition, and the content of glucose in the solution is measured, wherein the content of glucose is 6.2 percent; the reaction was then terminated. The pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5, and the mass of the reaction solution was made the initial mass (about 824g) by supplementing sterilized purified water.
(4) And (2) placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate with active carbon, concentrating, crystallizing and drying to obtain 211.2g of a white rhamnose finished product, wherein the yield is 124.7%, and the powder contains a large amount of glucose.
Example 11
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. 358g of sugar solution is weighed, when purified water is naturally cooled to 52 ℃, the sugar solution is dissolved in the purified water and uniformly stirred, so that the sugar solution is completely dissolved to form a reaction sugar solution, the mass percentage of glucose in the reaction sugar solution is 16%, and the sum of the mass percentages of glucose and rhamnose is 35%.
(2) Adjusting pH of the reaction sugar solution to 5.5 with calcium hydroxide, placing the reaction sugar solution in a 52 deg.C water bath, and oxygenating with oxygen at 1L/min.
(3) Adding 12mL of glucose oxidase (1.00g/mL, 20 ℃) and 12mL of catalase (1.06g/mL, 20 ℃) into the reaction sugar liquid respectively to obtain reaction liquid, and reacting the reaction liquid for 48 hours at 52 ℃ in an oxygenated state (oxygen introduction amount is 1L/min); measuring the content of glucose in the reaction solution, wherein the content of glucose is 3.8%; the reaction solution is continuously reacted for 24 hours under the same condition, and the content of glucose in the solution is measured, wherein the content of glucose is 3.4 percent; then the reaction solution is continuously reacted for 24 hours under the same condition, and the content of glucose in the solution is measured, wherein the content of glucose is 3.2 percent; continuously reacting the reaction solution for 24h under the same condition, and measuring the content of glucose in the reaction solution, wherein the content of glucose is 3.1%; the reaction was then terminated. The pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5, and the mass of the reaction solution was made the initial mass (about 828g) by supplementing sterilized purified water.
(4) And (2) placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate with active carbon, concentrating, crystallizing and drying to obtain 187.3g of a white rhamnose finished product, wherein the yield is 110.6%, and the powder contains a large amount of glucose.
Example 12
The embodiment provides a process method for purifying rhamnose from a glucose-containing sugar solution, which comprises the following steps:
(1) 446g of purified water is weighed, heated to 100 ℃ and kept for 30 min. 358g of sugar solution is weighed, when purified water is naturally cooled to 52 ℃, the sugar solution is dissolved in the purified water and uniformly stirred, so that the sugar solution is completely dissolved to form a reaction sugar solution, the mass percentage of glucose in the reaction sugar solution is 16%, and the sum of the mass percentages of glucose and rhamnose is 35%.
(2) Adjusting pH of the reaction sugar solution to 5.5 with calcium hydroxide, placing the reaction sugar solution in a 52 deg.C water bath, and oxygenating with oxygen at 1L/min.
(3) Adding 20mL of glucose oxidase (1.00g/mL, 20 ℃) and 10mL of catalase (1.06g/mL, 20 ℃) into the reaction sugar liquid respectively to obtain reaction liquid, and reacting the reaction liquid for 48 hours at 52 ℃ in an oxygenated state (oxygen introduction amount is 1L/min); measuring the content of glucose in the reaction solution, wherein the content of glucose is 3.9%; the reaction solution is continuously reacted for 24 hours under the same condition, and the content of glucose in the solution is measured, wherein the content of glucose is 3.2 percent; then the reaction solution is continuously reacted for 24 hours under the same condition, and the content of glucose in the solution is measured, wherein the content of glucose is 3.1 percent; continuously reacting the reaction solution for 24h under the same condition, and measuring the content of glucose in the reaction solution, wherein the content of glucose is 3.0%; the reaction was then terminated. The pH of the reaction solution was adjusted with calcium hydroxide during the reaction to maintain the pH at 5.5, and the mass of the reaction solution was made the initial mass (about 834g) by supplementing sterilized purified water.
(4) And (2) placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, collecting filtrate, decoloring the filtrate with active carbon, concentrating, crystallizing and drying to obtain 186.2g of a white rhamnose finished product, wherein the yield is 109.9%, and the powder contains a large amount of glucose.
The technical contents of the present invention are further illustrated by the examples, so as to facilitate the understanding of the reader, but the embodiments of the present invention are not limited thereto, and any technical extension or re-creation based on the present invention is protected by the present invention.
Claims (10)
1. The preparation method of rhamnose is characterized by comprising the following steps:
s1, dissolving the sugar solution in water to obtain a reaction sugar solution;
the sugar solution is a mixed solution containing rhamnose and glucose; the total mass of rhamnose and glucose in the reaction sugar solution is less than or equal to 35 percent;
s2, adjusting the pH value of the reaction sugar solution to 5.5-6.0 by using calcium hydroxide, then placing the reaction sugar solution in an environment with the temperature of 50-55 ℃, and continuously oxygenating the reaction sugar solution;
s3, adding glucose oxidase and catalase into the reaction sugar solution to obtain reaction liquid, and reacting the reaction liquid for 40-48 hours at 50-55 ℃ in an oxygenated state; adjusting the pH value of the reaction liquid with calcium hydroxide during the reaction process to keep the pH value at 5.5-6.0, and supplementing water to make the mass of the reaction liquid be 75-125% of the initial mass;
the addition amount of the glucose oxidase and the catalase is 2-3% of the mass of the reaction sugar solution;
and S4, placing the reaction solution at room temperature, neutralizing the reaction solution with calcium hydroxide until the pH value is 7.0, filtering, decoloring the filtrate with activated carbon, and concentrating and crystallizing to obtain rhamnose.
2. The method for preparing rhamnose according to claim 1, wherein the water is purified water that has been sterilized.
3. The method for preparing rhamnose according to claim 2, wherein the sterilization treatment is heating purified water to 100 ℃ and keeping the temperature for more than 30 min.
4. The method for preparing rhamnose according to claim 3, wherein in step S1, after the water is heated to 100 ℃ and kept warm for 30min, when the temperature is naturally cooled to 50-55 ℃, sugar solution is added into the water.
5. The method of claim 1, wherein the pH of the reaction sugar solution is adjusted to 5.5 with calcium hydroxide in step S2.
6. The method for preparing rhamnose according to claim 1, wherein in step S2, the reaction sugar solution is placed in an environment at 52 ℃.
7. The method for preparing rhamnose of claim 6, wherein in step S2, the reaction sugar solution is placed in a water bath at 50-55 ℃.
8. The method of producing rhamnose according to claim 1 wherein, in steps S2 and S3, oxygen is added to the reaction sugar solution or the reaction solution, and the amount of oxygen introduced per kg of the reaction sugar solution or the reaction solution is 1L/min or more.
9. The method for preparing rhamnose according to claim 1, wherein in step S3, the addition amounts of glucose oxidase and catalase are 2.5% of the mass of the reaction sugar solution.
10. The method for preparing rhamnose according to any one of claims 1 to 9, wherein in step S1, the sum of the mass of rhamnose and glucose in the reaction sugar solution is 25 to 35% of the mass of the reaction sugar solution.
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|---|---|---|---|---|
| CN114858940A (en) * | 2022-04-27 | 2022-08-05 | 浙江新银象生物工程有限公司 | Method for measuring content of L-rhamnose |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5077206A (en) * | 1987-11-20 | 1991-12-31 | Unilever Patent Holdings B.V. | Process for preparing l-rhamnose |
| CN102080106A (en) * | 2010-12-09 | 2011-06-01 | 山东西王生化科技有限公司 | Double enzyme method for preparing zinc gluconate |
| CN102102115A (en) * | 2010-12-09 | 2011-06-22 | 山东西王生化科技有限公司 | Method for preparing calcium gluconate and isomaltooligosaccharide simultaneously with crystalline glucose mother liquor |
| CN104450806A (en) * | 2014-11-27 | 2015-03-25 | 江南大学 | Method for preparing sodium gluconate with enzymatic method |
-
2020
- 2020-10-15 CN CN202011101481.7A patent/CN112125935A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5077206A (en) * | 1987-11-20 | 1991-12-31 | Unilever Patent Holdings B.V. | Process for preparing l-rhamnose |
| CN102080106A (en) * | 2010-12-09 | 2011-06-01 | 山东西王生化科技有限公司 | Double enzyme method for preparing zinc gluconate |
| CN102102115A (en) * | 2010-12-09 | 2011-06-22 | 山东西王生化科技有限公司 | Method for preparing calcium gluconate and isomaltooligosaccharide simultaneously with crystalline glucose mother liquor |
| CN104450806A (en) * | 2014-11-27 | 2015-03-25 | 江南大学 | Method for preparing sodium gluconate with enzymatic method |
Non-Patent Citations (3)
| Title |
|---|
| 李艳，等: "葡萄糖氧化酶及其应用", 《食品工程》 * |
| 肖芳，等: "穿龙薯蓣皂甙酸解液中鼠李糖的提取", 《化学与生物工程》 * |
| 高旭东，等: "L-鼠李糖的制备及检测方法研究进展", 《安徽农业科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114858940A (en) * | 2022-04-27 | 2022-08-05 | 浙江新银象生物工程有限公司 | Method for measuring content of L-rhamnose |
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