A kind of toxin protein and its purification method

A toxin protein, purification method technology, applied in chemical instruments and methods, peptide preparation methods, organic chemistry and other directions, can solve the problems of incomplete purification, no protein, irreversible protein adsorption, etc., to achieve strong practicability and simple operation steps , the effect of reducing adverse effects

A toxin protein, purification method technology, applied in chemical instruments and methods, peptide preparation methods, organic chemistry and other directions, can solve the problems of incomplete purification, no protein, irreversible protein adsorption, etc., to achieve strong practicability and simple operation steps , the effect of reducing adverse effects

CN103497242BActive Publication Date: 2016-01-20HUNAN NORMAL UNIVERSITY

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  • A kind of toxin protein and its purification method
  • A kind of toxin protein and its purification method
  • A kind of toxin protein and its purification method

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Embodiment 1

[0029] A toxin protein, extracted from the egg grains of the spider spider, the molecular weight is 28690, and its N-terminal sequence is Glu-Ser-Ile-Gln-Thr-Ser-Thr-Tyr-Val-Pro-Asn-Thr -Pro-Asn-Gln-Lys-Phe-Asp-Tyr-Glu-Val-Gly-Lys-Asp-Tyr-. The toxicity is slightly lower than some protein or polypeptide toxoids reported in the literature. The toxin protein can cause a series of poisoning symptoms in cockroaches and mice through intraperitoneal injection, and can inhibit tetrodotoxin-insensitive voltage-gated sodium ion channels on nerve cell membranes.

[0030] The purification method of the toxin protein specifically comprises the following steps:

[0031] The first step: the preparation of egg grain extract, homogenate the egg grains of the spider spider, take the supernatant as the extract after centrifugation, extract again after sedimentation and grinding, repeat 2-5 times, combine the extract, and pass through After freeze-drying or concentration, the egg extract is ob...

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Abstract

The invention provides a toxin protein which is separated and purified from latrodectus tredecimguttatus eggs, wherein the N-terminal sequence is Glu-Ser-Ile-Gln-Thr-Ser-Thr-Tyr-Val-Pro-Asn-Thr-Pro-Asn-Gln-Lys-Phe-Asp-Tyr-Glu-Val-Gly-Lys-Asp-Tyr-, and the molecular weight is 28,690. Intraperitoneal injection can induce a series of toxic symptoms of cockroaches and mouse; and a tetrodotoxin-resistant voltage-gated sodium ion channel on the nerve cell membrane can be inhibited. The invention also provides a purifying method of the toxin protein, which specifically comprises preparation of an egg extract, a molecular sieve chromatography step, an ion-exchange chromatography step and an inverse phase-high performance liquid chromatography step; and by introducing a C4 column with low hydrophobicity for purification, the steps are simple, and the purifying effect is good.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a toxin protein extracted from the ovum grains of the spider spider and a purification method thereof. Background technique [0002] L. tredecimguttatus, commonly known as black widow spider or black venomous spider, is a medium-sized spider that belongs to the phylum Arthropoda, Arachnida, Aranea, Ball Spideridae, and the genus Arachnida in taxonomy. . The venomous spider is one of the most poisonous spiders known in the world. Different from other poisonous animals, there is toxin not only in the venom of its venomous glands, but also in other tissues of the body (such as legs, abdominal tissues, etc.) and There are also toxic components in the eggs. It is of great theoretical and practical significance to study the toxic components outside the venom glands of A. [0003] Relevant foreign researchers have systematically and in-depth research on the venomous toxins of the spider ...

Claims

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Application Information

Patent Timeline
20 Jan 2016
Publication
CN103497242B
IPC
C07K14/435; C07K1/20; C07K1/18; C07K1/16
CPC
C07K14/43518
Inventors
王贤纯; 梁宋平