New way for synthesizing acetyl coenzyme A and derivative product thereof using glycolic aldehyde

Abstract

The invention discloses a new way for synthesizing acetyl coenzyme A and a derivative product thereof using glycolic aldehyde. The method comprises reaction that glycolic aldehyde reacts with 3-glyceraldehyde phosphate under enzyme catalysis to generate 5-phosphoric acid arabinose, wherein the enzyme is selected from aldolase, transaldolase and isozyme thereof and mutate enzyme. The method is relatively high in catalytic rate; the theoretical yield of carbon of a reaction route is 100%; the carbon loss avoided; G3P, an enzyme and a coenzyme all can be recycled; the reaction efficiency is relatively high; and the cost is reduced. In addition, the coenzymes RpiA and Rpe from a pentose phosphate pathway are high in affinity with a substrate and high in catalytic activity. According to the method, more obvious advantages are developed in the production processes, of in vitro continuous polyenzyme catalysis, fed-batch fermentation and continuous fermentation, capable of controlling the substrate level.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for synthesizing acetyl coenzyme A and derivatives thereof by using glycolaldehyde. Background technique [0002] Acetyl coenzyme A (AcCoA) is a key intermediate in the synthesis of essential biological compounds, including polyketides, fatty acids, isoprenoids, alkaloids, vitamins, and amino acids, among others. Metabolites derived from AcCoA are primary and secondary metabolites, which include compounds of industrial utility. AcCoA can be generated from acetyl phosphate (AcP) (for example, it can be catalyzed by phosphoacetyltransferase Pta (EC 2.3.1.8)), and then generate a series of biological products based on AcCoA platform, which are widely used in biocatalysis and other fields. The synthesis methods of AcP are: [0003] Patent application WO2015 / 144447A1 discloses a phosphoketolase (EC 4.1.2.9, fructose-6-phosphate phosphoketolase EC4.1.2.22) or sulfoacetal...

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Problems solved by technology

[0008] The reaction rate of the preparation method of above-mentioned acetyl phosphate is lower, and the conversion number K of phosphoketolase is for fructose cat Only 0.1 / s, the efficiency is very low, and the long reaction time (18h) will test the stability of the enzyme and product

In addition, the third step, the cracking of D-erythrose into glycolaldehyde is a reversible process, and its reaction rate is affected by the concentration of glycolaldehyde after cracking, making it difficult to accumulate high concentrations of glycolaldehyde in the system, which will lead to the formation of acetyl phosphate The speed is difficult to guarantee

The acetyl phosphate generated by the second enzymatic reaction is likely to affect the yield of the last enzymatic reaction due to product inhibition. If the second enzymatic reaction generates acetyl phosphate and removes it immediately, it will complicate the entire preparation process.

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