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Method for producing plant sterol ester by immobilized whole-cell enzyme catalysis in solvent-free system

A phytosterol ester, whole-cell technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, immobilized on/in organic carriers, etc., can solve poor solubility and bioavailability, small solubility, difficult and other problems, to achieve the effect of saving equipment investment, high recovery rate, and avoiding enzyme loss

Inactive Publication Date: 2010-06-02
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many literatures report that sterols play a role in preventing cardiovascular diseases, because it can effectively reduce the level of harmful low-density cholesterol (LDL), but the small solubility of free sterols in water and oil limits its application, and phytosterols are used in applications There are the following problems: 1) the crystalline form makes its solubility and bioavailability in the human body relatively poor; 2) it is insoluble in water, and its solubility in oil is also very small, which makes it difficult to apply to the food industry

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A method for catalyzing the production of phytosterol esters by immobilized whole-cell enzymes in a solvent-free system, comprising the steps of:

[0026] 1) Preparation of immobilized whole-cell enzyme:

[0027] (1) Commercially available Rhizopus stolonifer (Rhizopus stolonifer) was prepared into 10 2 Spore suspension, get Rhizopus stolonifer (Rhizopus stolonifer) spore suspension, subsequent use;

[0028] (2) Shake flask medium preparation: according to the mass percentage of each component: full-fat soybean powder (crushed after drying, passing through 30 mesh) 2.0%, rapeseed oil (commercially available) 0.3%, dipotassium hydrogen phosphate 1.2% %, Ammonium Sulfate 0.3%, Magnesium Sulfate 0.3%, Sodium Dihydrogen Phosphate 0.4%, Water 95.5%; choose full-fat soy flour, rapeseed oil, dipotassium hydrogen phosphate, ammonium sulfate, magnesium sulfate, sodium dihydrogen phosphate and water Mix to obtain a shaker flask culture medium and set aside;

[0029] (3) Polyur...

Embodiment 2

[0034] A method for catalyzing the production of phytosterol esters by immobilized whole-cell enzymes in a solvent-free system, comprising the steps of:

[0035] 1) Preparation of immobilized whole-cell enzyme:

[0036] (1) Commercially available Rhizopus stolonifer (Rhizopus stolonifer) was prepared into 10 3 Spore suspension, get Rhizopus stolonifer (Rhizopus stolonifer) spore suspension, subsequent use;

[0037] (2) Shake flask medium preparation: according to the mass percentage of each component: full-fat soybean powder (crushed after drying, passing through 40 mesh) 2.0%, rapeseed oil (commercially available) 0.5%, dipotassium hydrogen phosphate 1.2 %, Ammonium Sulfate 0.3%, Magnesium Sulfate 0.3%, Sodium Dihydrogen Phosphate 0.4%, Water 95.3%; choose full-fat soy flour, rapeseed oil, dipotassium hydrogen phosphate, ammonium sulfate, magnesium sulfate, sodium dihydrogen phosphate and water Mix to obtain a shaker flask culture medium and set aside;

[0038] (3) Polyure...

Embodiment 3

[0043] A method for catalyzing the production of phytosterol esters by immobilized whole-cell enzymes in a solvent-free system, comprising the steps of:

[0044] 1) Preparation of immobilized whole-cell enzyme:

[0045] (1) Commercially available Rhizopus stolonifer (Rhizopus stolonifer) was prepared into 10 3 Spore suspension, get Rhizopus stolonifer (Rhizopus stolonifer) spore suspension, subsequent use;

[0046] (2) Shake flask medium preparation: according to the mass percentage of each component: full-fat soybean powder (crushed after drying, passed through 30-50 mesh) 2.0%, rapeseed oil (commercially available) 0.5%, dihydrogen phosphate Potassium 1.2%, Ammonium Sulfate 0.3%, Magnesium Sulfate 0.3%, Sodium Dihydrogen Phosphate 0.4%, Water 95.3%; choose full-fat soybean flour, rapeseed oil, dipotassium hydrogen phosphate, ammonium sulfate, magnesium sulfate, sodium dihydrogen phosphate Mix it with water to get a shaker flask culture medium for later use;

[0047] (3) P...

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Abstract

The invention relates to a method of producing phytosterol esters, or a phytosterol esters production method with immobilized whole cell enzyme catalysis in a solvent free system. The invention is characterized in that the method includes the following steps: 1) the preparation of the immobilized whole cell enzyme; 2) the esterification: adding sterol and the immobilized whole cell enzyme in oleicacid, evening promose oil or edible soybean oil and putting the mixture in a water bath constant temperature oscillator for the esterification reaction of 46-50h at the temperature of 40-60 DEG C andthe rotation speed of 100-300r / min to obtain a reaction product after the esterification. The weight ratio of the oleic acid, the evening promose oil or the edible soybean oil to the sterol is 3-7 to1. The immobilized whole cell enzyme is that: (1) the oleic acid, the evening promose oil or the edible soybean oil; (2) sterol, which is 1.0-1.7 percent of the weight sum of the two; (3) the centrifugal separation and the drying: implementing the centrifugal separation on the reaction product after the esterification. The upper liquid after the centrifugation is a phytosterol esters product. Theinvention has the advantages of simple manufacturing process, low cost, repeated use and environmental protection and can be applied to the industrialized production.

Description

technical field [0001] The present invention relates to a method for producing phytosterol esters. Background technique [0002] Phytosterols are a natural product with physiological functions. Many literatures report that sterols play a role in preventing cardiovascular diseases, because it can effectively reduce the level of harmful low-density cholesterol (LDL), but the small solubility of free sterols in water and oil limits its application, and phytosterols are used in applications. There are the following problems: 1) the crystalline form makes its solubility and bioavailability in the human body relatively poor; 2) it is insoluble in water, and its solubility in oil is also very small, which makes it difficult to apply to the food industry. Since phytosterol esters have good fat solubility, the conversion of phytosterols and fatty acids into oil-soluble phytosterol esters can solve the limitation of application in the food industry. In addition, since oleic acid and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P33/00C12N11/08C12R1/845
Inventor 胡小加陈洪江木兰董绪燕刘昌盛魏芳张银波黄凤洪
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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