Method for enzymatic synthesis of theaflavin TF by virtue of tea polyphenol oxidase isoenzyme PPO1

Abstract

The invention discloses a method for enzymatic synthesis of theaflavin TF by virtue of tea polyphenol oxidase isoenzyme PPO1. The method is the optimization of a technique and a technology, which is capable of achieving the oriented enzymatic synthesis of a single theaflavin component TF through an enzymatic reaction with a shaking table as well as subsequent steps of extracting, centrifuging, concentrating, freeze-drying and the like by taking tea polyphenol oxidase isoenzyme PPO1 as an enzyme source, catechins EC and EGC components in tea polyphenol a major reaction substrates and a buffer solution at a certain pH value as a reaction system. The method overcomes shortcomings that multiple theaflavin components are generated simultaneously, a technology for separating and purifying the theaflavin components is complex, the preparation of a high-purity theaflavin component is quite difficult and the like in a conventional process which carries out the enzymatic synthesis of theaflavin by virtue of crude enzyme fluid of tea polyphenol oxidase; the method achieves a technological innovation that a single product is generated from enzymatic synthesis and a side product is avoided at reaction end, and the method avoids a technological bottleneck in subsequent separation and purification of the theaflavin component, so as to lay the theoretical and practical foundation for the green, safe and efficient large-scale industrial production and utilization of the theaflavin component.

Description

technical field [0001] The invention belongs to the field of bioengineering preparation, and relates to a preparation method of theaflavin TF, in particular to a tea polyphenol oxidase isoenzyme PPO1 enzyme liquid in a tea polyphenol solution reaction system to synthesize theaflavin TF directional enzymatically Methods. Background technique [0002] Theaflavins (theaflavins, TFs) are found in black tea with polyhydroxy benzotropolone structure mixture, accounting for 0.3%-1.5% of the dry weight of black tea, and play a key role in the color, aroma and quality of black tea. Current studies have shown that theaflavins have a variety of pharmacological effects related to human health, and they all play a role in preventing cardiovascular and cerebrovascular diseases, delaying aging, enhancing immune function, preventing dental caries, anti-tumor, antibacterial and virus-killing, etc. Known as "brain gold", it is one of the natural products with the most obvious efficacy, the m...

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Problems solved by technology

[0008] The technical problem to be solved by the present invention is: to directly extract and prepare theaflavins from tea leaves and prepare theaflavins by chemical oxidation to form the theaflavins mixture according to the above-mentioned prior art, polyphenol oxidase isozymes have complex composition and various enzymatic synthesis products, tea Due to the large differences in biological activity of flavin components, high cost of purification and preparation of theaflavin components, low degree of industrialization, and cumbersome operations, a method for enzymatically synthesizing single theaflavins using tea polyphenol oxidase isoenzyme PPO1 is provided. The method of the flavin component TF, using the specific tea polyphenol oxidase isoenzyme PPO1, to form the theaflavin component TF by catalyzing the oxidation of tea polyphenol solution conversion

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