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Method for preparing hydroxytyrosol by combining enzymatic catalysis and high-temperature hydrolysis

A technology of hydroxytyrosol and high-temperature hydrolysis, which is applied in the fields of biochemistry and medical and health care products, can solve the problems of high corrosion, high synthesis cost, and troublesome neutralization, and achieves the effects of cost saving and yield improvement.

Active Publication Date: 2017-09-29
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to its complicated preparation process, the price of commodity hydroxytyrosol is generally more expensive
When using acid and alkali to prepare hydroxytyrosol, because acid and alkali hydrolysis have high requirements on equipment, high corrosiveness, and troublesome neutralization, the small molecule sodium chloride or calcium sulfate generated after neutralization is easy to absorb small molecule hydroxytyrosol , making subsequent separation and purification more difficult
Although there are a small amount of reports on the synthesis of hydroxytyrosol by chemical methods, the cost of synthesis is high and the yield is low

Method used

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  • Method for preparing hydroxytyrosol by combining enzymatic catalysis and high-temperature hydrolysis
  • Method for preparing hydroxytyrosol by combining enzymatic catalysis and high-temperature hydrolysis
  • Method for preparing hydroxytyrosol by combining enzymatic catalysis and high-temperature hydrolysis

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Experimental program
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Effect test

Embodiment 1

[0029] Get concentration and be that 50mL of oleuropein solution of 10.0g / L is placed in the Erlenmeyer flask of 125mL, add the cellulase (Novozyme, β-glucosidase) that is rich in high activity β-glucosidase of 10FPU / g oleuropein The ratio of enzyme activity to filter paper enzyme activity is not less than 25), enzymolysis at 50°C for 12h, then raise the temperature to 90°C, and continue hydrolysis for 36h. After the enzymolysis, the enzymolysis solution was centrifuged at 8000r / min for 10min. After centrifugation, the filtrate was passed through a 0.22 μm filter membrane, and after dilution, HPLC detection and analysis showed that the degradation rate of oleuropein was 100%, and the yield of hydroxytyrosol was 88.90% (the content was equivalent to 2.53g / L).

Embodiment 2

[0031]Measure the oleuropein solution 50mL that concentration is 12.5g / L and place in the conical flask of 125mL, add the cellulase (Novozyme, β-glucosidase that is rich in the high activity β-glucosidase of 60FPU / g oleuropein The ratio of glucosidase activity to filter paper enzyme activity is not less than 25), hydrolyze at 50°C for 12h, then increase the temperature to 90°C, and continue hydrolysis for 36h. After the enzymolysis, the enzymolysis solution was centrifuged at 8000r / min for 10min. After centrifugation, the filtrate was passed through a 0.22 μm filter membrane, and after dilution, HPLC detection and analysis showed that the degradation rate of oleuropein was 100%, and the yield of hydroxytyrosol was 83.89% (the content was equivalent to 2.99g / L).

Embodiment 3

[0033] Measure 50 mL of oleuropein solution with a concentration of 10.0 g / L and place it in a 125 mL conical flask, add 50 FPU / g oleuropein rich in cellulase (Novozyme, β-glucosidase) The ratio of glucosidase activity to filter paper enzyme activity is not less than 25), hydrolyze at 50°C for 1 hour, then increase the temperature to 90°C, and continue hydrolysis for 47 hours. After the enzymolysis, the enzymolysis solution was centrifuged at 8000r / min for 10min. After centrifugation, the filtrate was passed through a 0.22 μm filter membrane, and after dilution, HPLC detection and analysis showed that the degradation rate of oleuropein was 100%, and the yield of hydroxytyrosol was 82.38% (the content was equivalent to 2.35g / L).

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Abstract

The invention discloses a method for preparing hydroxytyrosol by combining enzymatic catalysis and high-temperature hydrolysis. The method comprises the following steps of performing enzymatic hydrolysis on oleuropein by cellulose which is rich in beta-glucosaccharase, wherein the using amount of cellulose is 10 to 60FPU / g of oleuropein; and directly controlling temperature to 80 to 100 DEG C without processing to continuously hydrolyze for 24 hours or more, and obtaining hydroxytyrosol solution, wherein the degradation rate of the obtained oleuropein is 95 percent or more, and the yield of the hydroxytyrosol is 80 percent or more. The hydroxytyrosol is prepared from an olive leaf extract by the method of combining enzymatic catalysis and high-temperature hydrolysis, reaction is performed through mild enzymatic catalysis, the using amount of enzyme is relatively small, cost is saved, the degradation rate of the obtained oleuropein is 95 percent or more, the yield of the hydroxytyrosol is 80 to 90 percent, and particularly the yield of the hydroxytyrosol is obviously improved, so that a foundation is laid for industrial production.

Description

technical field [0001] The invention relates to the fields of biochemistry and medical health products, and relates to a method for preparing hydroxytyrosol from natural product olive leaf extracts, in particular to a method for preparing hydroxytyrosol by combining enzyme catalysis and high-temperature hydrolysis. Background technique [0002] Olive (Olea europaea L.), also known as olean, belongs to the evergreen tree of Oleaceae Oleaceae. Mediterranean countries, such as Spain, Greece, Italy, Tunisia, etc. are its concentrated production areas, and the planting area accounts for about 95% of the global planting area. . my country began to introduce olive trees on a large scale in the 1960s, mainly distributed in the central and western regions such as Gansu and Sichuan, with an area of ​​more than 450,000 mu and a total of more than 5 million trees. About 3-5kg of olive leaves can be obtained for every 100kg of olive fruit picked. Currently olive leaves are generally bu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/22C07C37/055C07C39/11
CPCC07C37/0555C12P7/22C07C39/11
Inventor 余世袁刘敏勇强连之娜朱均均
Owner NANJING FORESTRY UNIV
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