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Method for producing D-mannose

A technology of mannose and fructose, which is applied in the field of D-mannose production, can solve the problems that the output and production efficiency need to be improved urgently, and the output of D-mannose cannot meet the production demand, so as to achieve the effect of increased output and efficient production

Active Publication Date: 2019-02-12
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the yield of D-mannose prepared by microbial fermentation is still far from meeting the production demand, and the yield and production efficiency need to be improved urgently

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The fermentation culture of embodiment 1 Pseudomonas (Pseudomonas sp.) SK 27.016

[0022] (1) Seed cultivation

[0023] Seed medium: ammonium chloride 6g / L, disodium hydrogen phosphate 5g / L, potassium dihydrogen phosphate 0.5g / L, magnesium chloride 0.04g / L, sodium sulfate 0.5g / L, D-fructose 12g / L, deionized Prepare water and adjust the pH to 7.0;

[0024] Seed culture conditions: select well-cultured single colony SK 27.016 on the plate at 25-37°C, shaker speed 200rpm, and culture in the seed medium for 14-20h.

[0025] (2) Fermentation culture

[0026] Fermentation medium (in g / L): Ammonium chloride 6g / L, disodium hydrogen phosphate 5g / L, potassium dihydrogen phosphate 0.5g / L, magnesium chloride 0.04g / L, sodium sulfate 0.5g / L, D- Fructose 12g / L, glucose 6g / L, deionized water preparation; adjust pH 7.0.

[0027] Fermentation conditions: inoculum size 10%, temperature 35-37°C, shaker speed 180-200rpm, when the glucose in the fermentation medium is consumed, add 2g / L ...

Embodiment 2

[0048] Example 2 Cell transformation to produce D-mannose

[0049] Suspend the cells of Example 1, Comparative Example 1, and Comparative Example 2 with pH 7.5 phosphate buffer solution to OD 600 =2.5, then add 100 g / L of D-fructose, and convert for 2 hours at a temperature of 40°C. 41g / L, 32g / L, 28g / L D-mannose can be obtained respectively.

[0050] After the transformation solution was refrigerated and centrifuged, the supernatant containing D-mannose was collected, desalted and decolorized by strong acidic cation and weakly basic anion exchange resin in series, and then passed through DTF-Ca 2+ type ion exchange resin for separation and purification. As determined by high performance liquid chromatography, the purity of the isolated D-mannose can reach 98%.

Embodiment 3

[0051] Example 3 Cell transformation to produce D-mannose

[0052] Suspend the thalline prepared in Example 1 with pH 7.5 phosphate buffer solution to OD 600 = 2.0, then add 100 g / L D-fructose, convert for 2 hours at a temperature of 40° C., and obtain 39 g / L D-mannose.

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Abstract

The invention discloses a method for producing D-mannose, and belongs to the technical field of food biology. A culture method of Pseudomonas sp. SK 27.016 is optimized, D-fructose and glucose are taken as a mixed carbon source, and glucose is replenished in a culturing process to obtain a high-activity bacterial strain. When the bacterial strain is used for catalyzing the preparation of D-mannosefrom D-fructose, the yield can be improved greatly, and can be up to 41g / L, and the transformation rate is up to 41 percent. By adopting the method, the large-scale industrial preparation of the D-mannose can be made possible. The D-mannose produced is safe and reliable, is a functional product which has a great market potential, and can be widely applied to the industries of foods, makeups, medicine and the like. The method can efficiently produce the D-mannose, and is suitable for large-scale production.

Description

technical field [0001] The invention relates to a method for producing D-mannose, which belongs to the field of food biotechnology. Background technique [0002] D-Mannose, the only glyconutrient currently available clinically, is widely distributed in body fluids and tissues, especially in nerves, skin, testes, retina, liver and intestines. It is directly used to synthesize glycoproteins and participate in immune regulation. In addition, in recent years, it has been found that D-mannose can inhibit tumor growth and metastasis, increase the survival rate of cancer, and it has incomparable advantages in the treatment of urinary tract infection. D-Mannose, may be recommended as supplemental treatment for infants and children with a rare disorder called phosphorylated mannose isomerase deficiency, which is known as Carbohydrate Deficient Glycoprotein Syndrome Type 1b (CDGS Type Ib), Manifested as intestinal protein loss, liver disease, hypoglycemia, and coagulation disorders....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/02C12R1/38
CPCC12P19/02
Inventor 江波张涛
Owner JIANGNAN UNIV