Enzyme-catalyzed method for synthesizing isoquercitrin

A technology of isoquercitrin and enzymatic synthesis, applied in the direction of fermentation, etc., can solve the problems of poor solubility and low catalytic efficiency of α-L-rhamnosidase, and achieve the effects of mild reaction conditions and good industrial application prospects

Inactive Publication Date: 2011-12-21
上海诺能生物科技有限公司
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AI Technical Summary

Problems solved by technology

[0005] At present, the biosynthesis of isoquercitrin has not yet achieved large-scale industrial application. The reason is that in the traditional reaction system using water as the solvent, the solubility of most natural glycoside substrates such as rutin is poor, and the catalytic activity of α-L-rhamnosidase less efficient

Method used

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  • Enzyme-catalyzed method for synthesizing isoquercitrin

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The selected frequency is 10kHz and the intensity is 0.1w / cm 2 Ultrasound, 1min / h acts on the reaction system of α-L-rhamnosidase catalyzing the hydrolysis of rutin to synthesize isoquercitrin. With 0.01mol / L disodium hydrogen phosphate-citric acid buffer solution, prepare 0.01g / L rutin mother solution with pH=5. Add 0.1 g / L α-L-rhamnosidase solution to the rutin mother solution at a volume ratio of 1%, and shake and react at 10° C. at 50 r / min for 1 h. After the reaction was detected by HPLC, the conversion rate of the product isoquercitrin was increased by 0.2% compared with the method under the same conditions but without ultrasonic enhancement.

Embodiment 2

[0019] The selected frequency is 150kHz and the intensity is 1.5w / cm 2 The ultrasonic wave, 60min / h, acts on the reaction system in which α-L-rhamnosidase catalyzes the hydrolysis of rutin to synthesize isoquercitrin. Use 1mol / L disodium hydrogen phosphate-citric acid buffer solution to prepare 1 g / L rutin mother solution with pH=9. Add 100 g / L α-L-rhamnosidase solution to the rutin mother liquor at a volume ratio of 50%, and shake and react at 60° C. and 200 r / min for 48 hours. After the reaction was detected by HPLC, the conversion rate of the product isoquercitrin was increased by 2% compared with the method under the same conditions but without ultrasonic enhancement.

Embodiment 3

[0021] The selected frequency is 40kHz and the intensity is 0.28w / cm 2 The ultrasonic wave, 10min / h, acts on the reaction system in which α-L-rhamnosidase catalyzes the hydrolysis of rutin to synthesize isoquercitrin. With 0.2mol / L disodium hydrogen phosphate-0.1mol / L citric acid buffer solution, prepare 0.3g / L rutin mother solution with pH=7. Add 10 g / L α-L-rhamnosidase solution to the rutin mother solution at a volume ratio of 5%, and shake and react at 120 r / min at 40° C. for 20 h. After the reaction was detected by HPLC, the conversion rate of the product isoquercitrin was increased by 31% compared with the method under the same conditions but without ultrasonic enhancement.

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Abstract

The invention relates to an enhanced enzymic synthesis method for producing isoquercitrin, which comprises the following steps of: selecting ultrasonic waves with the intensity being 0 to 1.5w / cm<2> in a range being 10 to 150kHz for continuously acting on a reaction system for catalyzed hydrolysis of rutin to synthetize isoquercitrin through alpha-L-RhaR at 1 to 60min / h; using disodium hydrogen phosphate-citric acid buffer solution with the concentration being 0.01 to 1 mol / L for preparing 0.01 to 1g / L rutin mother liquid with pH being 5 to 9; adding 0.1 to 100g / L alpha-L-RhaR liquid in the rutin mother liquid according to the volume ratio being 1 to 50 percent; and carrying out oscillation reaction for 1 to 48h under the conditions of 10 to 60 DEG C and 50 to 200 r / min. The enhanced enzymic synthesis method has the advantages that the conversion rate of the isoquercitrin of the ultrasonic field enhancing enzyme synthesis products is higher than that of a method without ultrasonic enhancing under the same condition, in addition, the operation is simple and convenient, the condition is mild, and the speed and the intensity of the enzymatic reaction can be respectively accelerated and improved.

Description

technical field [0001] The invention belongs to the field of enzymatically synthesizing food additives, in particular to a method for enzymatically synthesizing isoquercitrin induced by an ultrasonic field. 2. Background technology [0002] Prior art: Isoquercitrin (Isoquercitrin), that is, quercetin-3-O-β-D-glucoside (Quercetin-3-glucose), is very rare in nature, and is a kind of Anti-tumor and other biologically active flavonoids advanced food additives and active ingredients of natural medicines are also the synthetic raw materials for new functional food additives EMIQ (Enzymatically Modified Isoquercitrin) in the world in recent years (Toxicology, 2010, 268(3): 213-218) . [0003] Patent CN 101985639 A uses rutin, which is widely distributed in nature and has abundant sources, as raw material, and isoquercitrin is prepared by glycosidase-catalyzed hydrolysis reaction, and an enzymatic hydrolysis process for the directional synthesis of isoquercitrin by biotransformatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/60
Inventor 王俊吴福安许晏张旭东
Owner 上海诺能生物科技有限公司
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