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Method for extracting L-type and E-type phytohemagglutinins (PHAs)

A plant lectin and L-type technology, which is applied in the field of extracting L-type and E-type plant lectins, can solve the problem of no great reference value for the extraction and separation of plant lectins, and achieve high purity and simple operation

Pending Publication Date: 2021-04-27
ZHENGZHOU IMMUNO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing patent CN103833839A discloses a C-type lectin and its preparation method, but the C-type lectin uses the hemolymph of Lepidoptera Cephalidae insects as the raw material, which has no great reference value for the extraction and separation of plant lectins
At present, there are no relevant reports on the extraction methods of the two subtypes of lectin PHA-L and PHA-E from beans in China.

Method used

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  • Method for extracting L-type and E-type phytohemagglutinins (PHAs)
  • Method for extracting L-type and E-type phytohemagglutinins (PHAs)
  • Method for extracting L-type and E-type phytohemagglutinins (PHAs)

Examples

Experimental program
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Embodiment 1

[0028] Example 1: Extraction and purification of L-type and E-type lectins

[0029] 1. Soak kidney beans in purified water at a mass volume ratio of 1:10 at 4°C for 12-24 hours, and then crush them with an electric crusher (the crushing temperature should not exceed 25°C, stop crushing when the crushing temperature reaches 25°C, and pre-cool until 4-25°C before crushing), after the crushing is completed, measure the volume of the crushed liquid with a graduated cylinder, and then stir at 4-25°C for 6-12h, centrifuge at 10,000rpm for 30min, and keep the centrifuged supernatant;

[0030] 2. After centrifugation, adjust the pH of the supernatant to 4.4-5.0, stir for 2 hours; centrifuge again at 10,000 rpm for 30 minutes, and keep the supernatant again;

[0031] 3. Slowly add solid ammonium sulfate to the supernatant component after centrifugation in step 2 to 35% saturation, stir for 30 minutes; centrifuge at 10000rpm for 30 minutes, and continue to retain the supernatant compone...

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Abstract

The invention relates to the technical field of plant extraction, and discloses a method for extracting L-type and E-type phytohemagglutinins (PHAs). According to the method, crude protein extraction is carried out through the steps of soaking, crushing, centrifuging, ammonium sulfate fractional precipitation and the like on beans, and then L-type and E-type PHAs are separated and purified through hydrophobic chromatography and ion exchange chromatography. The method is simple to operate, and the extracted L-type and E-type PHAs are high in purity and suitable for large-scale production of the L-type and E-type PHAs.

Description

technical field [0001] The invention relates to the technical field of plant extraction, in particular to a method for extracting L-type and E-type lectins. Background technique [0002] Phytohemagglutinin (Phytohemagglutinin PHA) is a sugar-containing protein extracted from the seeds of legumes. PHA is further divided into E-type and L-type, among which PHA-E type has erythrocyte agglutination; PHA-L type can stimulate lymphocyte transformation and is a stimulator of human lymphocyte mitosis, that is, under the action of PHA-L, it was originally in G0 Lymphocytes transform into lymphoblasts and undergo mitosis. [0003] There are also methods for extracting phytohemagglutinin PHA in the existing patented technology, and a variety of methods for extracting phytohemagglutinin from leguminous plants have been disclosed so far. For example, the Chinese invention patent application with the publication number CN101508730A discloses a method for extracting bean lectins, which u...

Claims

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Application Information

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IPC IPC(8): C07K14/42C07K1/36C07K1/34C07K1/30C07K1/20C07K1/18
CPCC07K14/42Y02A50/30
Inventor 任晓光岳试超周同喜赵巧辉李桂林张春鸽付豪苏测洋吴学炜付光宇杨增利
Owner ZHENGZHOU IMMUNO BIOTECH
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