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Application of lyase in preparation of oligomeric proanthocyanidins and method for preparing oligomeric proanthocyanidins through combination of steam explosion and lyase

A low-polymerization proanthocyanidin and high-polymerization proanthocyanidin technology is applied in the direction of fermentation, which can solve the problems of low substrate conversion rate, complex process, environmental pollution, etc., and achieve the effect of mild reaction conditions, high extraction rate and simple degradation process

Pending Publication Date: 2021-11-02
WEIHAI BAIHE BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The object of the present invention is to provide an application of lyase in the preparation of oligomeric proanthocyanidins and a method for preparing oligomeric proanthocyanidins by steam explosion combined with lyase, which overcomes the low extraction rate of grape seed proanthocyanidins and serious environmental pollution in the traditional production of oligomeric proanthocyanidins. The substrate conversion rate is low, the process is complicated, etc.

Method used

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  • Application of lyase in preparation of oligomeric proanthocyanidins and method for preparing oligomeric proanthocyanidins through combination of steam explosion and lyase

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Add 130-mesh grape seed powder into deionized water and soak for 35 minutes. Then, the treated grape seed powder was subjected to steam explosion treatment, maintained at a pressure of 3.0MPa for 105s, and then the pressure was released within 0.00087s to complete the explosion. After blasting, collect blasting samples. Add 60mL of petroleum ether to 3.00g of the exploded grape seed sample, and degrease for 5h under the shaking condition of 150rpm. After degreasing, petroleum ether was removed by evaporation under reduced pressure to obtain a sample of defatted grape seed powder. Take 1.00g of degreased grape seed sample and place it in 20mL of acetone:ultrapure water:acetic acid solution with a volume ratio of 75:24.5:0.5, and ultrasonicate for 23min at 30°C and 20kHZ. After the sonication, shake for 1.5 h at room temperature under light-shielding conditions, the content of highly aggregated proanthocyanidins in the crude extract is 24.4%. After calculation, the ext...

Embodiment 2

[0042] Add 60-mesh grape seed powder into deionized water and soak for 20 minutes. Then, the treated grape seed powder was subjected to steam explosion treatment, maintained at a pressure of 2.5MPa for 90s, and then the pressure was released within 0.00087s to complete the explosion. After blasting, collect blasting samples. Add 20mL of petroleum ether to 1.00g of the exploded grape seed sample, and degrease for 2h under the shaking condition of 50rpm. After degreasing, petroleum ether was removed by evaporation under reduced pressure to obtain a sample of defatted grape seed powder. Take 1.00g of defatted grape seed sample and place it in 15mL of acetone:ultrapure water:acetic acid solution with a volume ratio of 72.5:24.5:3, and ultrasonicate for 15min at 20°C and 10kHZ. After the sonication, shake for 1 h at room temperature under light-shielding conditions, the content of highly aggregated proanthocyanidins in the crude extract is 21.4%. After calculation, the extractio...

Embodiment 3

[0044] Add 200-mesh grape seed powder into deionized water and soak for 50 minutes. Then, the treated grape seed powder was subjected to steam explosion treatment, maintained at a pressure of 3.5MPa for 120s, and then the pressure was released within 0.00087s to complete the explosion. After blasting, collect blasting samples. Add 100mL of petroleum ether to 5.00g of the blasted grape seed sample, and degrease for 8h under the shaking condition of 250rpm. After degreasing, petroleum ether was removed by evaporation under reduced pressure to obtain a sample of defatted grape seed powder. Take 3.00g of defatted grape seed samples and place them in 30mL of acetone:ultrapure water:acetic acid solution with a volume ratio of 73:26.5:0.5, and ultrasonicate for 30min at 35°C and 30kHZ. After the sonication, shake for 2 hours at room temperature under light-shielding conditions, the content of highly aggregated proanthocyanidins in the crude extract is 24.4%. After calculation, the...

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Abstract

The invention provides application of lyase in preparation of oligomeric proanthocyanidins and a method for preparing the oligomeric proanthocyanidins by combining steam explosion with the lyase, and relates to the technical field of biotransformation. The amino acid sequence of the lyase is one of SEQ ID NO. 1 to SEQ ID NO. 5. The method for preparing the oligomeric proanthocyanidins by combining steam explosion with lyase comprises the following steps: soaking grape seeds with water, and then carrying out explosion treatment to obtain an explosion sample; degreasing the obtained blasting sample to obtain a degreased sample; mixing the degreased sample with the extracting solution, and extracting to obtain a high polymer procyanidine crude extract; and mixing the obtained high polymer proanthocyanidins crude extract with lyase, and reacting under an anaerobic condition to obtain the oligomeric proanthocyanidins. The method for preparing the oligomeric proanthocyanidins has the advantages of simple degradation process of the oligomeric proanthocyanidins, mild reaction conditions, small environmental pollution, high yield of the oligomeric proanthocyanidins and the like.

Description

technical field [0001] The invention relates to the technical field of biotransformation, in particular to an application of a lyase in preparing oligomeric proanthocyanidins and a method for preparing oligomeric proanthocyanidins by combining steam explosion with lyase. Background technique [0002] Grape seeds account for 3-6% of the total weight of grapes. They are the main by-products of industries such as wine and grape beverages, and are rich in natural resources. According to statistics, China's annual grape production reaches more than 10 million tons, which are mainly used in the production of wine and grape beverages, while the remaining grape seeds cannot be fully utilized, resulting in a huge waste of resources. Grape seeds contain a large amount of proanthocyanidins (content 3.9%). Proanthocyanidins in grape seeds mainly exist in the outer integument and are tightly wrapped by lignin, protein, cellulose, and hemicellulose. In addition, proanthocyanidins also bin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P17/16
CPCC12P17/162
Inventor 袁其朋张杰王文雅程磊雨刘新力刘旭东李静马荣荣石晔
Owner WEIHAI BAIHE BIOTECH
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