Method for preparing pharmaceutical dextran and levulose by bi-enzyme coupled multistage membrane separation

A technology for separation of dextran and membrane, applied in the direction of fructose production, sugar production, application, etc., can solve the problems of complicated separation of follow-up products, low yield of target products, and affecting the safe use of medicinal dextran, so as to facilitate industrial promotion and application , high added value, and the effect of reducing production costs

Inactive Publication Date: 2013-08-28
GUANGXI ZHUANG AUTONOMOUS REGION CENT FOR ANALYSIS & TEST RES
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in China, there is a method of fermenting sucrose with bacteria or dextran sucrase to prepare dextran first, and then adding dextran-containing fermentation serum or dextranase to prepare dextran with different molecular weights, but the separation of subsequent products is more complicated. , and use traditional separation methods such as gauze filtration and ethanol fractionation to refine the product, which may affect the safe use of medicinal dextran, and the preparation method has a large room for improvement
In addition, the reaction between dextran sucrase and sucrose will quickly generate dextran with a molecular weight greater than 1 million in a short period of time. Although this method is outstanding in the conversion rate of sucrose, because the enzymatic hydrolysis of dextranase is random, the target product can be obtained rate is not high

Method used

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  • Method for preparing pharmaceutical dextran and levulose by bi-enzyme coupled multistage membrane separation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Dextransucrase was fermented by Leuconostoc enterococci, the fermentation broth was centrifuged at 12,000r / min and 4°C for 20min, and the supernatant was obtained. The enzyme activity was detected by DNS method. Dextranase comes from Amano Amano Enzyme Trading (Shanghai) Co., Ltd. According to the enzyme activity assay method provided by Amano and combined with the actual situation of the present invention, the assay conditions were slightly modified to measure the enzyme activity of dextranase. It was determined that the enzyme activity of dextran sucrase was 5 U / mL, and the enzyme activity of dextranase was 22,000 U / mL.

[0023]Use sterile water to prepare 1M sucrose solution as the substrate mother solution, and then the final concentration of the substrate is 0.6M, and the final concentration of the enzyme is 0.5U / mL (dextran sucrase) and 0.05U / mL (dextranase). Add the two kinds of enzyme liquids (i.e. keep the ratio of the enzyme activity of dextran sucrase and dex...

Embodiment 2

[0025] The acquisition of dextran sucrase and dextranase and the determination of enzyme activity are the same as in Example 1.

[0026] Use sterile water to prepare 1M sucrose solution as the substrate mother solution, and then the final concentration of the substrate is 0.4M, and the final concentration of the enzyme is 0.5U / mL (dextran sucrase) and 0.05U / mL (dextranase). Add the two kinds of enzyme liquids (i.e. keep the ratio of the enzyme activity of dextran sucrase and dextranase in the reaction system at 10:1), the preparation of the reaction liquid is diluted with sterile water, and 4L of the reaction liquid is prepared at one time. The reaction temperature was 24°C and the pH was 6.3. The reaction liquid is placed in the barrel at the front end of the membrane separation, and the first-stage ultrafiltration membrane module with a molecular weight cut-off of 50,000 is started, and the outlet pressure is adjusted to 0.03-0.15MPa (about 5-15psi, or the membrane Separati...

Embodiment 3

[0028] The acquisition of dextran sucrase and dextranase and the determination of enzyme activity are the same as in Example 1.

[0029] Use sterile water to prepare 1M sucrose solution as the substrate mother solution, and then the final concentration of the substrate is 0.2M, and the final concentration of the enzyme is 0.5U / mL (dextran sucrase) and 0.05U / mL (dextranase). Add the two kinds of enzyme liquids (i.e. keep the ratio of the enzyme activity of dextran sucrase and dextranase in the reaction system at 10:1), the preparation of the reaction liquid is diluted with sterile water, and 4L of the reaction liquid is prepared at one time. The reaction temperature was 25°C and the pH was 6.5. The reaction liquid is placed in the barrel at the front end of the membrane separation, and the first-stage ultrafiltration membrane module with a molecular weight cut-off of 70,000 is started, and the outlet pressure is adjusted to 0.03-0.15MPa (about 5-15psi, or the membrane Separati...

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Abstract

The invention discloses a method for preparing pharmaceutical dextran and levulose by bi-enzyme coupled multistage membrane separation, which comprises the following steps: simultaneously adding dextran sucrase and dextranase into a sucrose solution used as a substrate to form a bi-enzyme / sucrose reaction system; and after the reaction product is separated by a multistage membrane system composed of hyperfiltration membranes and a nanofiltration membranes, directly obtaining the pharmaceutical dextran in the hyperfiltration membrane separation stage, and obtaining the high-purity levulose solution in the last nanofiltration membrane separation stage, wherein the enzymes and macromolecular dextran contained in the trapped solution of the first-stage hyperfiltration membrane return to the bi-enzyme / sucrose reaction system to participate in the reaction again, thereby implementing cyclic utilization of the enzymes. The invention has the advantages of lower cost, environment friendliness, high safety and high efficiency, is simple to operate, implements oriented adjustable production of high-quality pharmaceutical dextran with target molecular weight, and obtains the high-purity high-added-value levulose byproduct.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals and biomass separation, in particular to a method for preparing medicinal dextran and fructose through double-enzyme coupled multi-stage membrane separation. Background technique [0002] Dextran is an exopolysaccharide produced by bacteria (such as Leuconostoc mesenteroides), and its main chain is composed of D-glucose units linked by α(1→6) glycosidic bonds. Dextran with different molecular weights has many clinical uses, such as being used as a plasma substitute, dredging microvessels, and preventing and treating thrombosis. "Chinese Pharmacopoeia" (2010 edition) includes three kinds of dextran and its derivative drugs including Dex-70 (average molecular weight about 70,000), Dex-40 (average molecular weight about 40,000), Dex-20 (average molecular weight about 20,000) There are 12 kinds in total. At present, most domestic preparations of dextran adopt the process of "strain fermentation-eth...

Claims

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Application Information

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IPC IPC(8): C12P19/18C12P19/14C12P19/04C12P19/02C08B37/02C13K11/00
Inventor 黄艳陈山浦媛媛邹青松姚晓麦卢安根莫建光
Owner GUANGXI ZHUANG AUTONOMOUS REGION CENT FOR ANALYSIS & TEST RES
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