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Method for preparing neohesperidin by enzymic method

A technology of neohesperidin and hesperidin, applied in the field of enzymatic preparation of neohesperidin, which can solve the problems of high cost of rhamnosidase and rhamnosyltransferase, increased difficulty in product separation and purification, and complex hydrolyzed products , to achieve the effect of saving separation and purification cost, low cost and wide source of raw materials

Active Publication Date: 2013-07-31
MEIZHOU GOLDEN POMELO HEALTH TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The defect of this method is that the hydrolyzate is complex, mainly including the mixture of hesperetin-7-glucoside and hesperetin, and the mixture can only be converted into neohesperidin under the action of rhamnosyltransferase in the hydrolysis system Hesperetin-7-glucoside, the conversion rate is too low, the difficulty of product separation and purification increases, resulting in higher cost
At the same time, the high cost of rhamnosidase and rhamnosyltransferase also greatly limits their application in industrial production.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Take 0.25 g of Sepabeads EC-EP carrier (80-120 mesh), soak in water for 24 hours, filter out, and set aside. Add to 1.25 ml rhamnosidase solution, add 1 ml of 1.4 M potassium dihydrogen phosphate buffer solution with a pH of 8, add the above-mentioned treated carrier, and shake at a constant temperature of 26±1°C for 72 hours. The carrier after the immobilized enzyme was filtered, washed with distilled water until the eluate was free of protein, and then washed three times with 0.02 M phosphate buffer at pH 8, and the enzyme activity was 150 U / g.

Embodiment 2

[0022] Take 0.25 g of 80-120 mesh Sepabeads EC-HA carrier filtered out after soaking in water for 24 hours, add 1.25 ml of 4% glutaraldehyde solution, shake for 30 min, filter out the carrier and add it to 1.25 ml of rhamnosyltransferase solution, Add 1 ml of 0.02 M potassium dihydrogen phosphate buffer solution with a pH of 8, and shake the reaction system at a constant temperature of 26±1°C for 12 hours. Filter, wash with distilled water until the eluate is free of protein, then wash three times with 0.02 M phosphate buffer with pH 8, and the enzyme activity is 250 U / g.

Embodiment 3

[0024] In Reactor 1, prepare 5×10 with 80% methanol -3 M Hesperidin and 3×10 -3 M CuCl 2 Solution, heated and reacted in a water bath at 40 °C for 1 h, distilled off methanol under reduced pressure to obtain an aqueous solution of hesperidin copper complex with a concentration of 1.5 g / L. Add the immobilized rhamnosidase prepared in Example 1, 45 °C, pH 4.0, react for 1 h, the raw materials are completely converted into hesperetin-7-glucoside copper complex and rhamnose. Separation through the 400-mesh sieve at the bottom of Reactor 1, the immobilized enzyme remained in Reactor 1; the reaction solution entered into Reactor 2, added the immobilized rhamnosyltransferase prepared in Example 2, and reacted at 35°C for 2h, orange Cortex-7-glucoside and rhamnose were completely converted to neohesperidin copper complex. Separation through the 400-mesh screen at the bottom of the reactor, the immobilized rhamnosyltransferase remains in reactor 2, the solution enters reactor 3, a...

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PUM

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Abstract

The invention discloses a method for preparing neohesperidin by an enzymic method, comprising the following steps: performing coordination between hesperidin and metal ions and dissolving in water, and realizing conversion from hesperidin to neohesperidin under the action of immobilized rhamnosidase and immobilized rhamnoside transferase. The method provided by the invention is adopted to solve the problem that production cost of neohesperidin widely used in medical, functional food and its cosmetic fields is high, and simultaneously opens up a new approach for recycling of industrial waste orange peel and high value-added utilization of hesperidin.

Description

technical field [0001] The invention relates to a method for preparing neohesperidin by enzymatic method. Background technique [0002] Neohesperidin belongs to dihydroflavonoids, and is usually distributed in sour orange, citrus aurantium, citrus aurantium, and immature pomelo, with a higher content in citrus aurantium. Pharmacological studies have shown that neohesperidin has antibacterial, anti-inflammatory, anti-allergic, anti-tumor and anti-melanin deposition effects, so it is widely used in medicine, functional food and cosmetics. Neohesperidin is also an important raw material for the synthesis of new sweetener neohesperidin dihydrochalcone. Neohesperidin dihydrochalcone is an important sweetener with intense sweetness, 1800 times sweeter than sucrose, refreshing taste and long-lasting aftertaste. Neohesperidin dihydrochalcone has been approved as a feed additive, food additive and cosmetic raw material in the European Union and the United States; in 1997, it was li...

Claims

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Application Information

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IPC IPC(8): C12P19/60
Inventor 周金林黄宝华卢宇靖
Owner MEIZHOU GOLDEN POMELO HEALTH TECH CO LTD
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