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Method for yeast cell to biosynthesizing 1,6- fructose sodium diphosphate

A kind of sodium fructose diphosphate, biosynthesis technology, applied in the fields of bioengineering and microbial fermentation, can solve the problems of low biosynthesis efficiency, unfavorable simple operation, cumbersome microbial cultivation, etc., to achieve improved synthesis efficiency, easy operation, and synthetic efficiency high effect

Inactive Publication Date: 2008-12-24
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Most of the transformation and synthesis methods reported now use mold mycelia as the microorganisms for transformation. The cultivation of such microorganisms is cumbersome and time-consuming, and it is not conducive to the simplification of operation. At the same time, the biosynthesis efficiency is not high.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Select the strain of Saccharomyces cerevisiae, culture the cells on a shaker at 30°C and 150r / min (medium: wort with a sugar content of 12 Berlin, pH 6.5, 100mL in a 500mL Erlenmeyer flask), collect the cells for 24 hours, and freeze them in J-6M large-capacity Yeast cells were obtained by centrifuging with a centrifuge (BECKMAN) at 10000 r / min at 4°C for 30 minutes.

[0033] Take 1 g of yeast cells obtained by centrifugation, add 11% glucose by mass, 5% sodium dihydrogen phosphate, and 30 mL of reaction solution with a pH of 7. The transformation temperature is 30°C, and the rotation speed is 150 r / min. Aqueous solution of chloroacetic acid was used to terminate the reaction (aqueous solution of trichloroacetic acid was used in 5 ml), and the concentration of sodium fructose 1,6-diphosphate in the fermented broth was measured as 4.79 g / L after treatment.

Embodiment 2

[0035] Saccharomyces cerevisiae strains were selected, cultured on a shaker at 30°C and 150r / min (medium: wort with a sugar content of 12 Berlin, pH 6.5, 100mL in a 500mL Erlenmeyer flask), and the cells were collected in a J-6M large-capacity flask for 24 hours. Yeast cells were obtained by centrifuging in a refrigerated centrifuge (BECKMAN) at 10,000 r / min at 4°C for 30 minutes.

[0036] Take 1 g of yeast cells obtained by centrifugation, add 30 mL of reaction solution with a mass percentage of 8% glucose, 5% sodium dihydrogen phosphate, and pH 8.4, transform at a temperature of 30°C, and rotate at a speed of 150 r / min. Trichloroacetic acid aqueous solution terminated the reaction (trichloroacetic acid aqueous solution consumption 5ml), and after treatment, the concentration of 1,6-sodium fructose diphosphate in the fermented liquid was 3.9g / L.

Embodiment 3

[0038] Select the strain of Saccharomyces cerevisiae, culture the cells on a shaker at 30°C and 150r / min (medium: wort with a sugar content of 12 Berlin, pH 6.5, 100mL in a 500mL Erlenmeyer flask), collect the cells for 24 hours, and freeze them in J-6M large-capacity Yeast cells were obtained by centrifuging with a centrifuge (BECKMAN) at 10,000 r / min at 4°C for 30 minutes.

[0039]Take 1 g of yeast cells obtained by centrifugation, add 9.7% glucose by mass, 5.8% sodium dihydrogen phosphate, and 30 mL of reaction solution with pH 8.8. The conversion temperature is 30 °C and the rotation speed is 150 r / min. After 6 hours of conversion, use 1 mol / L Trichloroacetic acid aqueous solution terminated the reaction (trichloroacetic acid aqueous solution consumption 5ml), and after treatment, the concentration of sodium fructose 1,6-diphosphate in the fermented liquid was measured as 5.07g / L.

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Abstract

The invention discloses a method for biologically synthesizing 1, 6-di- (2- ethylhexyl ) phosphoric acid fructose sodium of a yeast cell, comprising the following steps that: by taking 100mL of sterilizing reaction fluid to count, 1 to 5 grams of thalli which is obtained by pre-culture process is added to react for 4 to 10 hours at a temperature of between 25 and 30 DEG C and at a speed of 100 to 200r / min, trichloroacetic acid is added to stop the reaction, the clear solution of the 1, 6-di- (2- ethylhexyl ) phosphoric acid fructose sodium is obtained by centrifugalization; the thalli is cells which are obtained by pre-culturing for 12 to 36 hours, freezing and centrifuging of Saccharomyces cerevisiae; the pH value of the reaction fluid is between 5.0 and 9.0, the compositions of the reaction fluid in weight percentage are: 2 to 12 percent of glucose, 2 to 15 percent of sodium phosphate dibasic and the balance being water. The 1, 6-di- (2- ethylhexyl ) phosphoric acid fructose sodium synthesized by the method has simple process for culturing the yeast cells, shortens the time, and has simple converting system, easy operation and high FDP sodium converting efficiency.

Description

technical field [0001] The invention belongs to the fields of bioengineering and microbial fermentation, and in particular relates to a method for yeast cell biosynthesis of sodium fructose 1,6-diphosphate. Background technique [0002] 1,6-diphosphate fructose [Fructose-1,6-Diphosphate, referred to as FDP], the molecular formula is C 6 h 14 o 12 P 2 , often in the form of sodium, calcium, zinc and other salts. It is an intermediate product of sugar metabolism, soluble in water, necessary for human life activities, and plays an important role as an attenuator, biocatalyst and cell strengthening agent in the process of cell metabolism. [0003] Since Harden and Young discovered and clarified the basic characteristics of 1,6-diphosphate fructose in 1908, many scientists have studied the preparation and application of 1,6-diphosphate fructose. People such as Nenfeng synthesized FDP in the laboratory with the enzyme system of fresh baker's yeast or brewer's yeast in 1942, w...

Claims

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Application Information

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IPC IPC(8): C12P19/02C12R1/865C12R1/645
Inventor 陈启和章海锋何国庆刘婧傅明亮
Owner ZHEJIANG UNIV
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