Ferulic acid producing engineering strain, construction method and biotransformation method

Abstract

The invention discloses a ferulic acid producing engineering strain, a construction method and a biotransformation method. The provided engineering strain integrates a novel enzyme coupling system, a strong promoter is used to co-express a pandan alcohol oxidase gene, a coniferyl alcohol dehydrogenase gene and a coniferyl aldehyde dehydrogenase gene in escherichia coli, and eugenol is catalyzed through enzymatic reaction to generate ferulic acid. In the system, a xylose reductase is simultaneously coupled, xylose is reduced in a production process, meanwhile, oxidation type coenzyme nicotinamide adenine dinucleotide and nicotinamide-adenine dinucleotide phosphate are regenerated, and coenzyme balance of a ferulic acid biotransformation system is kept, so that the transformation rate and the yield of the product, i.e. the ferulic acid are improved. Compared with the prior art, the method has the advantages that an effective and simple implementation method for safe biotransformation of a medical intermediate product, i.e. the ferulic acid is provided, the transformation rate is up to over 95 percent, the concentration of the ferulic acid (end-product) is 11g / L, and the method has a relatively high industrialization 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 Recombinant Strains of Escherichia coli. Jorg Overhage, Alexander Steinbuchel, and Horst Priefert, Applied and Environmental Microbiology, 2003, 6569-6576) The enzymatic reaction of hydrogenase 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 this The two coenzymes are continuously consumed during the conversion process, and the reduced coenzyme produced 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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