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Bee venom peptide and separation and purification method thereof

A technology for separation and purification of melittin, applied in the biological field, can solve the problems of high reagent consumption cost, harsh purification conditions, low separation purity, etc., and achieve the effects of less loss of target substances, high separation purity, and simple purification steps.

Active Publication Date: 2022-07-22
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The current separation and purification methods of melittin need to be carried out under a series of process conditions such as specific pH impurity removal and ultrafiltration, and there are problems such as complex process, harsh purification conditions, high cost of reagent consumption, and low separation purity.

Method used

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  • Bee venom peptide and separation and purification method thereof
  • Bee venom peptide and separation and purification method thereof
  • Bee venom peptide and separation and purification method thereof

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Embodiment 1

[0054] (1) Solvent extraction

[0055] The crude bee venom was mixed according to the material-to-liquid ratio (crude bee venom: acetonitrile water extractant) 1g:1mL. Specifically, add 20% acetonitrile water extractant to the centrifuge tube, add crude bee venom to the 20% acetonitrile water extractant, vortex for 25min, centrifuge at 14000rpm for 20 minutes at 4°C, and take the supernatant. Repeat the vortex centrifugation step above and pool the supernatants.

[0056] (2) The first separation and purification of melittin

[0057] The supernatant was separated by reverse-phase C18 semi-preparative column, and 0.2% (volume percent) formic acid aqueous solution and acetonitrile system were used for gradient elution. The elution procedure was shown in Table 1. figure 1 The retention time is in the range of 14min-15min, and it is freeze-dried at low temperature to obtain melittin in white powder with a chromatographic purity greater than 90%, such as figure 2 shown.

[0058...

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Abstract

The invention discloses melittin and a separation and purification method thereof, and relates to the technical field of biology. The method comprises the following steps: carrying out primary reversed-phase C18 semi-preparative column separation on a bee venom crude extract to be separated and purified, carrying out gradient elution by taking a formic acid aqueous solution and acetonitrile as mobile phases, collecting an eluent, then freeze-drying the eluent, redissolving, and carrying out secondary reversed-phase C18 semi-preparative column separation on the redissolved solution, in the secondary reversed-phase C18 semi-preparative column separation, a trifluoroacetic acid aqueous solution and methanol are used as mobile phases for isocratic elution. The method can achieve the technical effects of high separation purity, less target substance loss and less organic solvent dosage without combining with an ultrafiltration process for impurity removal in a specific pH environment, and has the technical advantages of environmental friendliness, simple device, simplicity and convenience in process operation and suitability for industrial production.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a melittin and a method for separation and purification thereof. Background technique [0002] Peptides are composed of the carboxyl group (-COOH) of one amino acid and the amino group (-NH) of another amino acid. 2 ) products formed by dehydration condensation. Peptides play a key role in the growth and development, metabolism, disease, aging and death of natural organisms. In recent years, more and more research reports on peptides have been reported. Peptides can promote wound healing; anti-inflammatory, antibacterial, and antiviral infection; anti-aging, eliminating excess free radicals in the body; and anti-tumor, inducing tumor cell apoptosis. There are also many peptides with research value in agriculture worthy of our in-depth study. For example, plant peptides include wolfberry peptides, ginseng peptides, etc., and animal peptides include melittin, scorpion venom peptide...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K1/36C07K1/34C07K1/20
CPCC07K14/43572
Inventor 杨梦瑞王敏史浩川周剑
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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