Method for synthesizing 1,2,4-butanetriol by enzymatic reaction

A technology of enzymatic reaction and butanetriol, which is applied in the field of bioengineering, can solve the problems of low conversion rate and high production cost, and achieve the effects of improving capacity, avoiding many by-products, and shortening reaction time

Active Publication Date: 2018-02-09
NANJING UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The technical problem to be solved in the present invention is to provide a method for synthesizing 1,2,4-butanetriol by enzymatic reaction to solve the problem of high production cost and low conversion rate of D-1,2,4-butanetriol in the prior art The problem

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  • Method for synthesizing 1,2,4-butanetriol by enzymatic reaction

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Experimental program
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Effect test

Embodiment 1

[0035] The preparation of pure enzyme solution related to in vitro synthesis of 1,2,4-butanetriol, the specific steps are as follows:

[0036] (1) Introduce restriction sites BamHI and HindIII at the 5' end and 3' end of the D-xylose dehydrogenase xylB gene, respectively, perform double enzyme digestion on the xylB gene and PRSFDuet-1, and then connect the xylB gene to PRSFDuet- 1 on the carrier;

[0037] (2) at the 5' end and 3 end of the D-xylonate dehydratase xylD gene ’ Introduce restriction sites BamHI and XhoI into the end primers, perform double enzyme digestion on the xylD gene and PRSFDuet-1, and then connect the xylD gene to the PRSFDuet-1 vector;

[0038] (3) 5 in the benzoylformate decarboxylase mdlC gene ’ end and 3 ’ Restriction sites BamHI and SacI were introduced at the end; double restriction digestion was performed on the mdlC gene and PRSFDuet-1, and then the mdlC gene was connected to the PRSFDuet-1 vector;

[0039] (4) 5 in alcohol dehydrogenase adhP g...

Embodiment 2

[0043] D-1,2,4-butanetriol was synthesized in vitro, the specific steps are as follows:

[0044] Reaction system: the concentration of D-xylose dehydrogenase is 350U / mL, the concentration of D-xylose dehydratase is 250U / mL, the concentration of benzoylformate decarboxylase is 200U / mL, alcohol dehydrogenase The concentration is 300U / mL.

[0045] PBS (NaHPO 4 , NaH 2 PO 4 )50mM pH7.0, xylose 20g / L, MgCl 2 6mM, NAD + 0.5 mM, NADH 0.5mM, TPP 0.4mM; finally add PBS to 10mL, the reaction temperature is 37°C, the reaction time is 24h, and the product D-1,2,4-butanetriol is obtained.

[0046] According to the in vitro synthesis system of Example 2, 1,2,4-butanetriol was detected by high performance liquid chromatography, and the final yield was 12 g / L, and the yield was 85%.

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Abstract

The invention discloses a method for synthesizing 1,2,4-butanetriol by enzymatic reaction. The method is characterized in that D-xylose dehydrogenase, D-xylonic acid dehydratase, benzoylformate decarboxylase and alcohol dehydrogenase are taken as catalysts for catalyzing conversion of D-xylose into D-1,2,4-butanetriol in a reaction system. The reaction system contains the following components: PBS(NaHPO4 and NaH2PO4) with concentration being 50 mM, NAD<+> with concentration being 0.5 mM, NADH with concentration being 0.5mM and TPP with concentration being 0.4 mM. The capability of synthesizing butanetriol in vitro is improved by controlling the quantity of enzymes, and the adverse effects of multiple byproducts, complex subsequent separation process and the like caused by a microbial synthesis method are avoided.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a method for synthesizing 1,2,4-butanetriol by enzymatic reaction. Background technique [0002] D-1,2,4-Butanetriol is a colorless, odorless, transparent, viscous four-carbon polyol. Industrial D-1,2,4-butanetriol is an important organic synthesis intermediate, widely used in medicine, agriculture, cosmetics, papermaking, polymer materials, tobacco, military industry and other fields. The nitro compound of D-1,2,4-butanetriol has low impact sensitivity, good thermal stability, low toxicity, and good hygroscopicity. When mixed with other energetic plasticizers, it can significantly improve the Low temperature mechanical properties of base propellants. [0003] At present, the production of 1,2,4-butanetriol at home and abroad is mainly realized by chemical synthesis, but a series of disadvantages such as harsh reaction conditions, high cost, and many by-produ...

Claims

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Application Information

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IPC IPC(8): C12P7/18
CPCC12P7/18
Inventor 陈可泉高倩王昕胡社伟许娜娜欧阳平凯
Owner NANJING UNIV OF TECH
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