Application of helicase in directionally hydrolyzing rutin for biosynthesizing Quereetin

A biosynthesis and helicase technology, applied in the direction of fermentation, etc., can solve the problems of unrealized directional transformation of quercetin, no rutinoside bonds, etc., and achieve good industrial application prospects, high catalytic efficiency and specificity, and enzyme catalysis High conversion effect

Inactive Publication Date: 2010-11-10
北京优联宏远生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(2) Enzyme-catalyzed hydrolysis mainly uses glycosidase as a catalyst, such as the method for preparing isoquercetin and quercetin by enzymatically hydrolyzing rutin disclosed in CN03133636, using Sophora japonica as high-temperature aerobic bacteria or Aspergillus or Lorenzo Fermentation of Saccharomyces coccus to produce enzyme indu

Method used

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  • Application of helicase in directionally hydrolyzing rutin for biosynthesizing Quereetin
  • Application of helicase in directionally hydrolyzing rutin for biosynthesizing Quereetin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A. The free enzyme of helicase catalyzes the directional biosynthesis of quercetin by hydrolyzing rutin:

[0030] Rutin was dissolved in a pH 7 buffer to obtain a rutin substrate solution with a concentration of 0.005 g / L. The substrate solution was placed in a 50 mL Erlenmeyer flask with a stopper, the helicase crude enzyme preparation was added in a ratio of 50 g / L, the cork was tightly ground, and the enzymatic hydrolysis reaction was carried out in a rotary shaker at 120 rpm. The reaction temperature is 40°C, and the reaction time is 0.5h. After the reaction was completed, the molar conversion rate of the hydrolyzed quercetin detected by HPLC was 92%.

[0031] B. The immobilized enzyme of helicase catalyzes the directional biosynthesis of quercetin by hydrolyzing rutin:

[0032] Prepare 0.5%wt colloidal solution for immobilized enzyme with carrageenan, add 50g / L helicase crude enzyme powder, mix well, drop into cross-linking agent solution and solidify for 3 hours...

Embodiment 2

[0035] A. The free enzyme of helicase catalyzes the directional biosynthesis of quercetin by hydrolyzing rutin:

[0036] Rutin was dissolved in a pH3 buffer to obtain a rutin substrate solution with a concentration of 0.100 g / L. Place the substrate solution in a 50mL conical flask with a stopper, add the crude helicase preparation at a ratio of 0.1g / L, plug the cork tightly, and carry out the enzymatic hydrolysis reaction in a rotary shaker at 120rpm , the reaction temperature is 10°C, and the reaction time is 36h. After the reaction was completed, the molar conversion rate of the hydrolyzed quercetin detected by HPLC was 22%.

[0037]B. The immobilized enzyme of helicase catalyzes the directional biosynthesis of quercetin by hydrolyzing rutin:

[0038] Use gelatin to make 15%wt colloidal solution for immobilized enzyme, add 0.100g / L helicase crude enzyme powder, mix well, drop into crosslinking agent solution and solidify for 12h, and make square immobilized particles with ...

Embodiment 3

[0041] A. The free enzyme of helicase catalyzes the directional biosynthesis of quercetin by hydrolyzing rutin:

[0042] Rutin was dissolved in a pH 9 buffer to obtain a rutin substrate solution with a concentration of 0.025 g / L. Place the substrate solution in a 50mL conical flask with a stopper, add the crude helicase preparation at a rate of 0.5g / L, plug the cork tightly, and carry out the enzymatic hydrolysis reaction in a rotary shaker at 120rpm , the reaction temperature is 70°C, and the reaction time is 0.1h. After the reaction was completed, the molar conversion rate of the hydrolyzed product quercetin detected by HPLC was 23%.

[0043] B. The immobilized enzyme of helicase catalyzes the directional biosynthesis of quercetin by hydrolyzing rutin:

[0044] Prepare a 5%wt colloidal solution for immobilized enzymes with sodium alginate, add 0.5g / L helicase crude enzyme powder, mix well, drop into the crosslinking agent solution for 0.5h, and make spherical shapes with u...

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Abstract

The invention provides an application of helicase in directionally hydrolyzing rutin for biosynthesizing Quereetin. The method provided by the invention comprises the following steps: adding a crude helicase preparation in rutin substrate solution with the concentration of 0.005-0.100g/L, enabling the helicase concentration in the solution to be 0.1-50g/L and the pH value to be 3-9, performing enzymatic hydrolysis reaction under the conditions that the reaction temperature is 10-70 DEG C and the reaction time is 0.1-36h, and acquiring the hydrolyzed product Quereetin after reaction. The invention adopts the crude helicase preparation or immobilized helicase preparation to directionally catalyze and hydrolyze rutin for biosynthesizing Quereetin, thus having the advantages of wide sources of catalyst, easy preparation and low cost, and the helicase preparation has high stability, can be stored easily, has high catalytic efficiency and high specificity. Therefore, by utilizing the helicase, the production cost of the Quereetin can be greatly reduced, the enzymatic conversion rate is high and the product is specific.

Description

1. Technical field [0001] The invention belongs to the field of natural product biosynthesis, and specifically relates to an application method of helicase in hydrolyzing rutin to prepare quercetin. 2. Background technology [0002] Prior art: quercetin (Quercetin, 3, 5, 7, 3', 4'-pentahydroxyflavanone) is a kind of flavonoid aglycone compound widely distributed in nature, is a kind of excellent natural antioxidant, has expectorant Cough and asthma, anti-inflammatory, anti-allergic, antispasmodic, cardiotonic, lowering blood pressure, lowering blood fat, anti-arrhythmia, anti-platelet aggregation, anti-oxidation, anti-tumor, anti-virus and other extensive pharmacological effects. Therefore, quercetin has always been one of the hot compounds in the pharmaceutical industry, and a lot of research has been done on its physical and chemical properties, extraction and separation, pharmacological activity and clinical application at home and abroad. [0003] Quercetin, as an aglyc...

Claims

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Application Information

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IPC IPC(8): C12P17/06
Inventor 王俊吴福安王蒙张妍谢谢
Owner 北京优联宏远生物技术有限公司
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