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CN-116925786-B - Recombinant gleditsia sinensis saponin and preparation method thereof

CN116925786BCN 116925786 BCN116925786 BCN 116925786BCN-116925786-B

Abstract

The invention relates to the field of natural product separation and recombination, in particular to a recombinant gleditsia sinensis saponin and a preparation method thereof. The method comprises the steps of 1) extracting crude gleditsia sinensis saponin, 2) removing impurities, namely loading the crude gleditsia sinensis saponin aqueous solution into macroporous adsorption resin AB-8, eluting to remove impurities, 3) separating saponin components, namely eluting with 25-35% ethanol solution, 40-50% ethanol solution, 55-65% ethanol solution and 70-80% ethanol solution in sequence to respectively obtain S30, S45, S60 and S75 gleditsia sinensis saponin components, 4) hydrolyzing the S45 gleditsiae saponin components to obtain hydrolyzed saponin, and 5) recombining the gleditsia sinensis saponin components obtained in the steps 3) and 4) and the hydrolyzed saponin to obtain recombined gleditsiae saponin. The method can realize the purification of the saponin and improve the surface activity of the gleditsia sinensis saponin.

Inventors

  • CHEN DIANSONG
  • MA LING
  • JIANG JIANXIN
  • ZHU LIWEI
  • ZHANG FENGLUN

Assignees

  • 广州德谷个人护理用品有限公司
  • 北京林业大学

Dates

Publication Date
20260505
Application Date
20230706

Claims (13)

  1. 1. A preparation method of recombinant gleditsia sinensis saponin is characterized by comprising the following steps: 1) Extracting gleditsia sinensis saponin, namely extracting gleditsia sinensis pericarp with water to obtain a gleditsia sinensis saponin solution; 2) Impurity removal, namely loading the crude gleditsia sinensis saponin solution into macroporous adsorption resin AB-8, and eluting to remove impurities; 3) Separating saponin components, namely eluting with 25-35% ethanol solution, 40-50% ethanol solution, 55-65% ethanol solution and 70-80% ethanol solution in sequence to obtain S30, S45, S60 and S75 gleditsia sinensis saponin components respectively; 4) Hydrolyzing the S45 gleditsia sinensis saponin component to obtain hydrolyzed saponin; 5) And (3) recombining the saponin components, namely recombining the gleditsia sinensis saponin components obtained in the steps 3) and 4) and the hydrolyzed saponin to obtain recombined gleditsia sinensis saponin, wherein the gleditsia sinensis saponin component is one or more selected from the group consisting of an S30 gleditsia sinensis saponin component, an S45 gleditsia sinensis saponin component, an S60 gleditsia sinensis saponin component and an S75 gleditsiae saponin component.
  2. 2. The method for preparing recombinant gleditsia sinensis saponin according to claim 1, wherein in the step 1), deionized water is used as an extractant, and the solid-to-liquid ratio is 1:6-10.
  3. 3. The preparation method of the recombinant gleditsia sinensis saponin according to claim 2, wherein in the step 1), gleditsia sinensis pericarp is crushed and sieved by a 30-50 mesh sieve, deionized water is used as an extracting agent, the solid-liquid ratio is 1:6-8, and the raw gleditsia sinensis saponin aqueous solution is obtained through stirring, extracting and filtering.
  4. 4. The method for preparing recombinant gleditsia sinensis saponin according to claim 1, wherein in the step 2), deionized water and 5-15% ethanol solution are used for eluting sequentially.
  5. 5. The method for preparing recombinant gleditsia sinensis saponin according to claim 4, wherein in the step 2), the crude gleditsia sinensis saponin solution is loaded into macroporous adsorption resin AB-8, and is sequentially eluted by deionized water with 2-3 times of resin volume and 5-15% ethanol solution with 2-3 times of resin volume.
  6. 6. The method for preparing recombinant gleditsia sinensis saponin according to claim 1, wherein in the step 3), 30-35% ethanol solution, 45-50% ethanol solution, 60-65% ethanol solution and 75-80% ethanol solution are used for elution.
  7. 7. The method for preparing recombinant gleditsia sinensis saponin according to claim 6, wherein in the step 3), the 30-35% ethanol solution is 3-8 times of resin volume, the 45-50% ethanol solution is 8-12 times of resin volume, the 60-65% ethanol solution is 3-8 times of resin volume, and the 60-65% ethanol solution is 2-3 times of resin volume.
  8. 8. The method for preparing recombinant gleditsia sinensis saponin according to claim 1, wherein in the step 4), the S45 gleditsia sinensis saponin component is prepared into an aqueous solution with a concentration of 15-25%, the pH is adjusted to 1.5-2.5, and then the aqueous solution is hydrolyzed at a temperature of 75-85 ℃.
  9. 9. The method for preparing recombinant gleditsia sinensis saponin according to any one of claims 1-8, wherein in step 5), the ratio of the gleditsia sinensis saponin component to the hydrolyzed saponin is adjusted, so that the critical micelle concentration and the surface tension of the recombinant gleditsia sinensis saponin are adjustable.
  10. 10. A recombinant gleditsia sinensis saponin obtained by the process of any one of claims 1-9.
  11. 11. The recombinant gleditsia sinensis saponin according to claim 10, wherein said recombinant gleditsia sinensis saponin is a hydrolyzed saponin and a complex of one or more selected from the group consisting of S30 gleditsia sinensis saponin fraction, S45 gleditsia sinensis saponin fraction, S60 gleditsia sinensis saponin fraction and S75 gleditsiae saponin fraction.
  12. 12. The recombinant gleditsia sinensis saponin according to claim 11, wherein the mass ratio of the S30 gleditsia sinensis saponin component, the hydrolyzed saponin, the S60 gleditsia sinensis saponin component and the S75 gleditsia sinensis saponin component is 1-2:15-20:15-20:1.5-2.
  13. 13. The recombinant gleditsia sinensis saponin according to any one of claims 10-12, wherein the recombinant gleditsia sinensis saponin has a surface tension of 30-35 mN/m and a critical micelle concentration of 0.08-0.12 g/L.

Description

Recombinant gleditsia sinensis saponin and preparation method thereof Technical Field The invention relates to the field of natural product separation and recombination, in particular to a recombinant gleditsia sinensis saponin and a preparation method thereof. Background The gleditsia sinensis (L.) Schott belongs to the genus gleditsia of the family Leguminosae, is a typical ecological and economic useful tree species, is mainly distributed in areas such as China, southeast Asia, north America, south America and the like, and is widely planted in areas such as Shanxi, henan, shandong, guizhou, yunnan and the like of China. The gleditsia sinensis seed endosperm contains rich galactomannans, and the gleditsia sinensis polysaccharide has high economic and nutritional values due to high carbohydrate content, low protein content and low fat content. Meanwhile, the gleditsia sinensis galactomannan is similar to guar gum and locust gum polysaccharide in structure and function, and is a non-traditional industrial polysaccharide gum. Gleditsia sinensis galactomannans are often used as thickeners, binders, stabilizers and flocculants in a wide variety of industries such as food, daily chemicals, medicine, oil drilling and printing. While utilizing the Gleditsia galactomannans, we have found that the pericarp of Gleditsia contains a large amount of natural saponin. The saponin structure has the hydrophilic large polar groups such as sugar, ligand, organic acid and the like, while the aglycone structure is larger, has low polarity and lipophilicity, has the structural characteristics similar to a surfactant on the molecular structure, can generate lasting foam similar to a soap solution after being shaken by water solution, is a nonionic natural surfactant, and has good detergency. In the production process of natural saponin, the impurities are difficult to remove, the purification cost is high, partial saponin structure is unstable, and the saponin surface activity is required to be further improved. Disclosure of Invention In order to solve the technical problems, the invention provides recombinant gleditsia sinensis saponin and a preparation method thereof. In a first aspect, the present invention provides a method for preparing recombinant gleditsia sinensis saponin, comprising: 1) Extracting gleditsia sinensis saponin, namely extracting gleditsia sinensis pericarp with water to obtain a gleditsia sinensis saponin solution; 2) Impurity removal, namely loading the crude gleditsia sinensis saponin aqueous solution into macroporous adsorption resin AB-8, and eluting to remove impurities; 3) Separating saponin components, namely eluting with 25-35% ethanol solution, 40-50% ethanol solution, 55-65% ethanol solution and 70-80% ethanol solution in sequence to obtain S30, S45, S60 and S75 gleditsia sinensis saponin components respectively; 4) Hydrolyzing the S45 gleditsia sinensis saponin component to obtain hydrolyzed saponin; 5) And 4) recombining the saponin components, namely recombining the gleditsia sinensis saponin components obtained in the steps 3) and 4) and the hydrolyzed saponin to obtain the recombined gleditsia sinensis saponin. The invention firstly adopts a water extraction method to prepare a crude product of the natural gleditsia sinensis saponin, then the crude saponin water solution is subjected to macroporous adsorption resin AB-8 to remove impurities and separate the gleditsia sinensis saponin components, the separated saponin S45 is hydrolyzed and recombined with other saponin components in an optimized way, and the surface activity of the saponin is improved while the purification of the saponin is realized. Preferably, in the step 1), deionized water is used as an extracting agent, the solid-liquid ratio is preferably 1:6-10, preferably, the gleditsia sinensis fruit peel is crushed and passes through a 30-50 mesh sieve, the deionized water is used as the extracting agent, the solid-liquid ratio is 1:6-8, stirring extraction is carried out for 5-8 hours, and the crude gleditsia sinensis saponin aqueous solution is obtained through filtration. Further preferably, in the step 1), crude gleditsia sinensis saponin is extracted by crushing gleditsia sinensis pericarp, sieving with a 40-mesh sieve, taking deionized water as an extracting agent, stirring and extracting for 6-7 hours according to the solid-to-liquid ratio of 1:7-8, and filtering to obtain crude gleditsia sinensis saponin aqueous solution. In the invention, the water extraction method is adopted to treat the gleditsia sinensis peel, extract crude saponin and prepare the crude gleditsia sinensis saponin aqueous solution, thus not only having low extraction cost but also having high extraction rate of saponin. Preferably, in the step 2), deionized water and 5-15% ethanol solution are used for eluting sequentially, preferably, the crude gleditsia sinensis saponin solution is filled into macroporous adsorption resin AB-8, and deionized water w