Method for synthetizing 1,6-fructose diphosphate through bioconversion of lignocellulose biomass

A technology of lignocellulose and fructose diphosphate, applied in the field of bioengineering, can solve the problems of unsatisfactory industrial production, unstable ATP, expensive raw materials, etc., achieve good economic and social benefits, alleviate environmental pollution problems, and simple operation Effect

Inactive Publication Date: 2019-06-07
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method suffers from the need to add expensive ATP, although it can be regenerated in vitro, ATP is unstable at higher temperatures, so biocatalysis cannot be performed at high reaction temperatures
[0005] It can be seen that the current market potential of FDP is relatively large, but the biological synthesis of FDP still has problems such as expensive raw materials and low efficiency, which cannot meet the needs of industrial production.

Method used

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  • Method for synthetizing 1,6-fructose diphosphate through bioconversion of lignocellulose biomass
  • Method for synthetizing 1,6-fructose diphosphate through bioconversion of lignocellulose biomass

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1: Synthesis of fructose 1,6-diphosphate by biotransformation of rice straw

[0039] (1) Double hydrogen bond donor deep eutectic solvent (choline chloride: formic acid: acetic acid) pretreatment of rice straw

[0040] Take 100g of choline chloride: formic acid: acetic acid (1:1:1), add 10g of rice straw (mass ratio of deep eutectic solvent to rice straw is 10:1), stir at 140°C for 2h, then centrifuge the mixture , and washed the precipitate 3 times with deionized water. After drying to constant weight, sieve. Then treat with 0.5% sodium carbonate solution at 80° C. for 1.5 h, dry and sieve to obtain the rice straw combined with deep eutectic solvent and sodium carbonate.

[0041] (2) Dual-enzyme conversion of cellulose to synthesize cellobiose

[0042] Mix the pretreated rice straw with citric acid-sodium citrate buffer (pH 5.0, 100mM) at a solid-to-liquid ratio of 10%, and then add 50U / g of endocellulase and 50U / g of exocellulase Enzymatic hydrolysis, enzyma...

Embodiment 2

[0045] Example 2: Synthesis of fructose 1,6-diphosphate by biotransformation of rice straw

[0046] (1) Pretreatment of rice straw with new deep eutectic solvent (ethylamine hydrochloride: lactic acid)

[0047] Take 100g of ethylamine hydrochloride: lactic acid (1:1), add 10g of rice straw (mass ratio of deep eutectic solvent to rice straw is 10:1), stir at 140°C for 2h, then centrifuge the mixture, and use The precipitate was washed 3 times with deionized water. After drying to constant weight, sieve. Then treat with 0.5% sodium carbonate at 80° C. for 1.5 h, dry and sieve to obtain the rice straw combined with deep eutectic solvent and sodium carbonate.

[0048] (2) Dual-enzyme conversion of cellulose to synthesize cellobiose

[0049] Mix the pretreated rice straw with citric acid-sodium citrate buffer (pH 5.0, 100mM) at a solid-to-liquid ratio of 10%, and then add 50U / g of endocellulase and 50U / g of exocellulase Enzymatic hydrolysis, enzymatic hydrolysis on a water-bath...

Embodiment 3

[0052] Example 3: Synthesis of fructose 1,6-diphosphate by biotransformation of rice straw

[0053] (1) Double hydrogen bond donor deep eutectic solvent (choline chloride: formic acid: acetic acid) pretreatment of rice straw

[0054] Take 100g of choline chloride: formic acid: acetic acid (1:1:1), add 5g of rice straw (mass ratio of deep eutectic solvent to rice straw is 20:1), stir at 120°C for 3h, then centrifuge the mixture , and washed the precipitate 3 times with deionized water. After drying to constant weight, sieve. Then treat with 5% sodium carbonate at 50° C. for 6 hours, dry and sieve to obtain rice straw combined with deep eutectic solvent and sodium carbonate.

[0055] (2) Dual-enzyme conversion of cellulose to synthesize cellobiose

[0056] Mix the pretreated rice straw with citric acid-sodium citrate buffer (pH 5.0, 100mM) at a solid-to-liquid ratio of 5%, and then add 50U / g of endocellulase and 50U / g of exocellulase Enzymatic hydrolysis, enzymatic hydrolysi...

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Abstract

The invention discloses a method for synthetizing 1,6-fructose diphosphate through bioconversion of lignocellulose biomass, belongs to the technical field of bioengineering, and discloses a technologyfor synthetizing high added value 1,6-fructose diphosphate through the lignocellulose biomass. A dual-hydrogen-bond eutectic solvent is used as a pretreatment reagent to remove hemicellulose and lignin in biomass, so that the accessibility of cellulose can be improved; incision enzymes and excision enzymes of the cellulose are added, and the cellulose is converted into cellobiose; and cellobiosephosphorylase, plastidial phosphoglucomutase, phosphoglucose isomerase and ppi dependent phosphofructokinase are added for synthetizing the 1,6-fructose diphosphate. The lignocellulose biomass which is cheap and easy to obtain is used as a starting material, and a high-energy phosphoric acid compound is synthetized through bioconversion; and the method has the characteristics of being low in cost,mild in reaction, simple to operate and the like; and solving the problem of environmental pollution caused by a conventional treatment manner of straw is facilitated, and the method has excellent economic benefits and social benefits.

Description

technical field [0001] The invention relates to a method for biotransforming lignocellulosic biomass to synthesize fructose 1,6-diphosphate, belonging to the technical field of bioengineering. Background technique [0002] Fructose-1,6-diphosphate (FDP), also known as fructose 1,6-diphosphate, is an important metabolic intermediate product of the glycolysis pathway in organisms, and is usually widely used in medicine as a drug. The World Health Organization Collaborating Center for Pharmaceutical Statistics Methodology classifies it as "Other cardiac preparations" (ATC code: C01EB07). Its cytoprotective activity has been documented in severe conditions such as convulsions, septic shock, diabetic complications, hypothermia-induced injury, UV-induced skin damage, and other processes including apoptosis and excitotoxicity. Clinical data suggest that this compound is useful in the treatment of various ischemic conditions, eg, bypass surgery, myocardial infarction, and transplan...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): C12P19/18C12P19/02
Inventor贾鹏王显众倪晔许国超邢婉茹龚磊
OwnerJIANGNAN UNIV