Extraction method of efficient active enzymes of fig latex

A high-efficiency active enzyme and extraction method technology, applied in the field of high-efficiency active enzyme extraction from fig milk, can solve the problems of unfavorable industrial production of refined fig protease, restriction of fig protease application, complex production process, etc., to shorten the separation time and reduce enzyme activity The effect of reduction and high purity

Inactive Publication Date: 2015-10-21
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In recent years, there have been many reports on the extraction and application of ficin, but most of the crude enzymes used in the market are crude ficins. Because the extraction process of crude enzymes is simple and the production cost is low, hot air drying is generally used to obtain ficins. Vitality is only about 10000u / g; at the same time, due to the complex production process and high production cost of other refined ficinase, it is not conducive to the industrial production of refined ficinase, which also limits the application of ficinase in high-end medicine and health care products and beauty cosmetics. Industry application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Fresh latex was collected from the incision of fig branches, and the latex was centrifuged at 10,000 rpm for 30 minutes to remove gum and other insoluble components. The remaining clear liquid was natural latex, which was stored at 4°C for enzyme purification. All subsequent steps were carried out at 4°C to prevent enzyme degradation and depolymerization.

[0029] The protein content was determined by the Coomassie brilliant blue method, and bovine serum albumin was used as the standard curve.

[0030] For the detection of enzyme activity, tyrosine was used as the standard curve, casein was used as the substrate, and Folin’s phenol reagent was used for color reaction.

[0031] 1. Acetone precipitation method

[0032] Latex is first precipitated with acetone. The enzymes in the latex were precipitated in 20–40% (v / v) acetone solution, and then centrifuged at 10,000 rpm. The enzyme was vacuum-dried to remove acetone and dissolved in 10 mM Tris-HCl buffer (pH 8.0). Det...

Embodiment 2

[0040] The specific extraction steps are the same as those in Example 1, except that in Step 2, Sephadex G-50 column is used for filtration. The enzyme activity is 25600U / mg, and the final yield is 9.15%.

Embodiment 3

[0042] The specific extraction steps are the same as in Example 1, except that the Sephadex G-100 column is used for filtration in step 2. The enzymatic activity of the finally extracted ficin was 25650U / mg, and the final yield was 9.21%.

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PUM

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Abstract

The invention discloses an extraction method of efficient active enzymes of fig latex. The method comprises the steps that acetone precipitation, gel chromatography separation, anion chromatography and vacuum freeze-drying are sequentially conducted on the fig latex which is pretreated at the temperature of 4 DEG C to obtain efficient active enzyme powder. Due to the fact that pretreatment and heat exchange are conducted and the extraction process is conducted under a low-temperature condition, the influence of conditions such as long-term high temperature on reducing enzyme activity can be effectively reduced; by means of milling action of a colloid mill, fig enzymes are dissolved in material liquid as many as possible, and the subsequent extraction is facilitated. By means of continuous separation through a disc-type centrifuge and a tubular bowl centrifuge, clear fig protease material liquid is obtained. The vacuum freezing and drying technology is adopted, due to the fact that when water in materials becomes ice, the water directly sublimates to be discharged, it is guaranteed that drying is conducted on the enzymes in the low-temperature condition, and the loss of enzyme mass and reducing of the enzyme activity in the drying process are reduced.

Description

technical field [0001] The invention relates to the field of food. Specifically, it relates to a method for extracting high-efficiency active enzymes from fig milk. Background technique [0002] Fig (Ficus carica Linn.) is a flowering plant belonging to the genus Moraceae. It mainly grows in some tropical and temperate regions and is a subtropical deciduous small tree. There are currently 800 known varieties of figs, most of which are evergreen varieties, and only those that grow in temperate regions are deciduous varieties. The fruit is bulbous with a small hole in the tail, and the pollen is spread by wasps. Because of its good medicinal value, it is more and more popular. [0003] Ficin (Papain) widely exists in the stems, leaves and fruits of figs, and the content of immature fruit milk is the highest. It can be prepared by chemical methods (precipitation method) and biotechnology methods. Ficin is a plant protease, referred to as ficin, because of its good thermal s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/50
CPCC12N9/63C12Y304/22003
Inventor 孙锐
Owner QILU UNIV OF TECH
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