A ferulic acid production engineering strain, construction method and biotransformation method

Abstract

The invention discloses a ferulic acid production engineering strain, a construction method and a biotransformation method. The provided engineering strain integrates a new enzyme coupling system, and utilizes a strong promoter to make vanillin alcohol oxidase gene, coniferyl alcohol dehydrogenase The gene and coniferous aldehyde dehydrogenase gene were co-expressed in Escherichia coli, and then eugenol was catalyzed to generate ferulic acid through enzymatic reaction. The system is coupled with xylose reductase at the same time, reducing xylose during the production process, while regenerating the oxidized coenzyme nicotinamide adenine dinucleotide and nicotinamide adenine dinucleotide phosphate, maintaining the ferulic acid biotransformation system Coenzyme balance, thereby increasing the conversion rate and yield of the product ferulic acid. Compared with the current prior art, the present invention provides an effective and simple implementation method for the safe biotransformation of the pharmaceutical intermediate product ferulic acid, the conversion rate reaches more than 95%, and the concentration of the final product ferulic acid reaches 11g / L, which has a relatively high High industrial value.

Description

technical field [0001] The invention belongs to the technical field of production of pharmaceutical raw materials, and in particular relates to an engineering strain capable of catalyzing eugenol to generate ferulic acid, a construction method and a biotransformation method. Background technique [0002] The chemical name of ferulic acid is 4-hydroxy-3-methoxycinnamic acid, which is one of the derivatives of cinnamic acid. Because ferulic acid can be used as a precursor for microbial fermentation to produce vanillin, as well as its anti-oxidation, anti-thrombotic, blood lipid-lowering, and immune-regulating properties, ferulic acid and its derivatives are widely used in Lowering blood fat, preventing and treating coronary heart disease and cancer, antibacterial and anti-inflammatory, anti-mutation, promoting blood circulation, eliminating chloasma, and medical treatment of insomnia, backache and leg sinking, health care product production and food additives. [0003] Indust...

AI Technical Summary

Problems solved by technology

The extraction process of this traditional production method is complex, involving multiple steps of high energy consumption and heavy pollution, resulting in a large amount of acid-base waste and waste water; at the same time, due to the low content of ferulic acid in rice bran, the yield of the final extraction product is insufficient 0.2%, the above factors have greatly limited the development of ferulic acid application industry

(Highly Efficient Biotransformation of Eugenol to Ferulic Acid and Further Conversion to Vanillin in RecombinantStrains of Escherichia coli. Jorg Overhage, Alexander Steinbuchel, and Horst Priefert, Applied and Environmental Microbiology, 2003, 6569-6576) In the above reaction, the The enzymatic reaction needs the oxidized coenzyme nicotinamide adenine dinucleotide phosphate (NADP), and the enzymatic reaction of coniferyl aldehyde dehydrogenase needs the oxidized coenzyme nicotinamide adenine dinucleotide (NAD), so the two coenzymes It is continuously consumed during the conversion process, and the reduced coenzyme generated at the same time feedbacks the catalytic efficiency of the enzyme, resulting in a large accumulation of intermediate products coniferyl alcohol and coniferaldehyde, with low production capacity and no industrial production value

Adding corresponding coenzymes, namely NADP and NAD, during the production transformation process can effectively solve the problem of insufficient oxidized coenzymes and greatly improve the efficiency and yield of ferulic acid biotransformation, but due to the high price and low stability of such coenzymes, It is not feasible to add a large amount of coenzyme

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