Novel flavone reductase and application thereof

A technology of reductase and flavone, applied in the field of new flavone reductase and its application, can solve the problem of unclear reaction mechanism

Pending Publication Date: 2022-05-17
艾博锐克生物科技(山东)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the enzyme gene responsible for the key reaction of the first step in the degradation of flavonoids - the hydrogenation of C2-C3 double bonds to dihydroflavones has not been discovered and identified, and the internal reaction mechanism is not clear. This is the current understanding of microbial metabolism of flavones. An important gap in the class of compounds

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  • Novel flavone reductase and application thereof
  • Novel flavone reductase and application thereof
  • Novel flavone reductase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0148] Example 1 Discovery and verification of flavone reductase FLR

[0149] experimental method:

[0150] Human intestinal flora can metabolize flavonoids, and the key enzymes responsible for the initial reaction in the metabolic pathways of the two most important types of compounds—flavonoids and flavonols have not yet been discovered and identified ( figure 1 a). The possible candidate genes of flavone reductase were predicted by the method of comparative genomics, and the candidate genes were cloned into plasmids one by one and introduced into Escherichia coli BL21(DE3) for expression, and anaerobic fermentation was carried out in the medium supplemented with apigenin , to determine whether there is target flavonoid generation.

[0151] Experimental steps:

[0152] 1) We tested whether the reported flavonoid reductase - isoflavone reductase IFR can catalyze the reduction of flavones, and found that IFR can only specifically catalyze the reduction of isoflavones. It sh...

Embodiment 2

[0158] The biochemical characteristic analysis of embodiment 2 flavone reductase

[0159] experimental method:

[0160] A coupling reaction system based on flavin cofactors was constructed to realize the catalytic reaction of flavone reductase in vitro, and the catalytic efficiency of flavone reduction reaction and dihydroflavone oxidation reaction of the enzyme was detected at different pH values, different temperatures and different substrate concentrations. To analyze the enzymatic properties of FLR.

[0161] Experimental steps:

[0162] 1), according to the two characteristic absorption peaks of 380nm and 447nm that appear in the full-wavelength scanning result of enzyme extract ( figure 2 a), Preliminary judgment that FLR is a flavin-dependent oxidoreductase.

[0163] 2). A coupled reaction system based on flavin cofactors was constructed to analyze the enzymatic properties of FLR. Such as figure 2 As shown in b, the reaction system produces reduced flavin cofactor...

Embodiment 3

[0168] The universality of embodiment 3 flavone reductase substrates

[0169] experimental method:

[0170] Under the established enzyme catalytic system, different kinds of representative flavone substrates were selected to test the catalytic activity of flavone reductase.

[0171] Experimental steps:

[0172] We selected 14 representative flavonoids from the main classes of flavonoids (flavonoids, flavonols, isoflavones, anthocyanins and chromones) for enzyme activity testing.

[0173] Experimental results:

[0174] The results showed that FLR had catalytic activity for these several substances, showing good substrate adaptability ( image 3 ). Moreover, it was found through comparison that if more or larger substituent groups appear on the B ring of these flavones, the catalytic efficiency of FLR to the substrate will be reduced. For example, the ability of FLR to catalyze the reduction of chrysin, apigenin, and luteolin shows a decreasing trend, and the efficiency of ...

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Abstract

The invention relates to novel flavonoid reductase and application thereof, in particular to flavonoid reductase capable of catalyzing a flavonoid compound/flavonol compound to be reduced into a flavanone compound/flavanol compound. And catalyzing the flavanone compound/flavanonol compound to be oxidized into a flavonoid compound/flavanonol compound.

Description

technical field [0001] The invention relates to the field of biotechnology; more specifically, the invention relates to a new type of flavone reductase and its application. Background technique [0002] An important class of natural products in human food is flavonoids, which are the most common polyphenolic compounds in nature and widely exist in plant foods such as vegetables, fruits, and grains. Flavonoids have inhibitory and killing effects on a variety of bacteria in vivo and in vitro experiments, and have anti-inflammatory, anti-tumor, and anti-oxidative physiological activities on mammalian cells, and can be used as drugs for clinical treatment of patients with cardiovascular diseases . In the coexistence of flavonoids in the intestinal environment, intestinal bacteria have been subjected to antibacterial stress from flavonoids for a long time, and have evolved a variety of enzymes that can degrade flavonoids, making them undergo chemical catalytic processes such as ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N15/53C12P17/06
CPCC12N9/001C12P17/06
Inventor 不公告发明人
Owner 艾博锐克生物科技(山东)有限公司
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