Method for preparing 3, 4-dihydroxyphenethyl alcohol through biological enzyme catalysis

Abstract

The invention discloses a method for preparing 3, 4-dihydroxyphenethyl alcohol through bio-enzyme catalysis, which comprises the following steps: by taking escherichia coli fermentation liquor for inducing enzyme production as a raw material, centrifuging to obtain thalli, adding a buffer solution to suspend the thalli, introducing oxygen and the like, and adding a substrate p-hydroxyphenethyl alcohol and bio-enzyme catalysis to react to obtain the 3, 4-dihydroxyphenethyl alcohol. According to the method, the 3, 4-dihydroxyphenethyl alcohol is obtained through biological enzyme catalysis and biological synthesis by taking the p-hydroxyphenethyl alcohol as a substrate, the reaction condition is mild, no by-product is generated, and the residual quantity of the substrate is small; the catalytic efficiency of the biological enzyme is high and reaches 99%, and the yield of the product 3, 4-dihydroxyphenethyl alcohol reaches 13 g / L or above; the preparation method is green, environment-friendly and short in reaction period, can be applied to large-scale industrial production, and meets market requirements.

Description

technical field [0001] The invention relates to a method for preparing 3,4-dihydroxyphenethyl alcohol catalyzed by biological enzymes, belonging to the technical field of bioengineering. Background technique [0002] 3,4-Dihydroxyphenylethanol, also known as hydroxytyrosol, molecular formula C 8 h 10 o 3 , is a natural polyphenolic compound, found in olive oil and wastewater from olive oil processing, mainly esterified oleuropein can be hydrolyzed to obtain free 3,4-dihydroxyphenylethanol, studies have shown that 3 , 4-Dihydroxyphenylethanol has strong antioxidant activity. In addition, it also has a variety of biological and pharmacological activities, which can prevent osteoporosis, help diabetes, obesity and other diseases related to mitochondrial dysfunction. It can reduce the incidence of such diseases and is widely used in food industry, nutritional health care and cosmetics. Therefore, the extraction and preparation of 3,4-dihydroxyphenylethanol has certain practi...

AI Technical Summary

Problems solved by technology

[0003] 3,4-Dihydroxyphenylethanol, the traditional preparation method is to extract from olive oil or olive leaves, a large amount of flammable and explosive organic reagents such as ethyl acetate, absolute ethanol, dichloromethane, etc. are needed in the extraction process, and the extraction The yield is only 8.4%, the yield is low, and more by-products will be contained in the extraction process, resulting in lower purity of p-hydroxyphenethyl alcohol; the commonly used preparation method is generally prepared by synthesis, for example, Xiaoyan , Zhang Yang, etc. disclosed the synthesis method of hydroxytyrosol, reported that catechol and glyoxylic acid were used as raw materials to synthesize 3,4-dihydroxyphenylethanol through 3,4-dihydroxymandelic acid, and synthesized by four-step reaction 3,4-Dihydroxyphenylethanol was discovered, the total yield was 52.7%, and the yield was relatively low; there are other preparation methods using biological preparation methods, for example, a kind of engineering bacteria and its application disclosed in Chinese patent CN201710659225.1, reported co-express E. coli with L-amino acid oxidase, α-ketoacid decarboxylase, alcohol dehydrogenase, and NAD(P) reductase four enzymes to biosynthesize 2-phenylethanol, tyrosol, and hydroxytyrosol. Among them, The yield of 3,4-dihydroxyphenylethanol is 435mg / L, and the yield is relatively low; Chinese patent CN201710659225.1 discloses Escherichia coli expressing hydroxytyrosol and hydroxytyrosol glucoside and its construction method and application, and reports that the constructed Escherichia coli contains And be able to express ARO10, HpaBC and UGT genes, recombinant Escherichia coli tyrosine is expressed through related genes to obtain 3,4-dihydroxyphenylethanol, its yield can reach 401mg / L, and the yield is relatively low

Images