Preparing method for extracting polypeptide antibiotics from silkworm

A technology of polypeptide antibiotics and silkworm, which is applied in the fields of biochemistry, insect immunology, and microbiology. It can solve the problems of low expression efficiency of eukaryotic expression system in industrial production, inability to obtain expression products, and suicide of host microorganisms. The preparation process is simple and feasible. , Extensive sources of raw materials and easy operation

Inactive Publication Date: 2003-11-19
成都天创生物科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the toxicity of polypeptide antibiotics to prokaryotic cells (such as microorganisms) may cause host microorganisms to commit suicide and fail to obtain expression products, which limits their application in prokaryotic expression systems to a certain extent.
The lower expression efficiency of the eukaryotic expression system is also an obstacle to its industrial production
Express the polypeptide antibiotic gene in the form of fusion protein, although this shortcoming can be overcome, there is still the problem of few expression products

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] 1. Get 4 or 5 instar silkworm larvae to add food to the mulberry leaves sprayed with 2% or 3% or 5% or 7% or 8.5% or 9.5% concentration of E. coli solution, and add food regularly once a day for four consecutive days.

[0055] 2. While adding food, treat the larvae with 11 or 14 or 16 or 17 or 19 kHz ultrasonic waves, induce once a day, 4 hours each time, and repeat the induction 3 times at intervals of 21, 22 or 23 hours. The induced silkworm larvae are stored at a low temperature of 1° C. or 3° C. or 5° C. or 7° C. or 9° C. or 11° C. for use.

[0056] 3. Take out the induced larvae, add 2 to 3 times the volume of 0.2 mol / L phosphate buffer, pH 6.2, put them into a tissue masher homogenizer, and homogenize for 10 minutes. The obtained homogenate was centrifuged for 15 min, and the supernatant was collected and placed at 4°C for 12 hours.

[0057] 4. Treat the extract in a water bath at 40°C or 45°C or 50°C or 55°C or 60°C or 65°C or 75°C or 80°C for 25 minutes, then q...

Embodiment 2

[0064] 1, at the 5th instar stage of silkworm, feed the mulberry leaves that are sprayed with 3% or 4.5% or 5.5% or 7.5% or 8% or 9% concentration of nuclear polyhedron inactivation solution, regularly add food once a day until Silkworm spins silk.

[0065] 2. After the silkworm larvae form cocoons, treat them with ultrasonic waves of 10 or 10.5 or 13 or 15 or 17.5 or 18 kHz for 4 hours, induce once a day, and repeat the induction three times at intervals of 20 to 24 hours. The induced silkworm chrysalis is stored at a low temperature of 2° C., 4° C., 6° C., 8° C., 10° C. or 12° C. for use.

[0066] 3. The extraction method is the same as in Example 1.

[0067] Yield: 8 mg / kg silkworm chrysalis.

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PUM

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Abstract

A process for extracting polypeptide antibiotic from bombxy mori includes such steps as including polypeptide antibiotic in young silkworm of pupa, preparing extracting liquid, separating and purifying. Its advantages are simple process, low cost, broad antibacterial spectrum, and high purity.

Description

technical field [0001] The invention relates to the fields of insect immunology, biochemistry, microbiology and the like, and more specifically relates to a preparation method for extracting polypeptide antibiotics from silkworm bodies. Background technique [0002] The first antimicrobial peptide discovered in the world was discovered in 1980 by Swedish scientist G.Boman et al. 12 D. 31 ) induces the polypeptide with antibacterial activity produced by the chrysalis chrysalis, named Cecropins. In the following years, people successively discovered and isolated peptides with antibacterial activity from bacteria, fungi, amphibians, insects, higher plants, mammals and even humans. Since it was initially discovered that this type of active peptide has broad-spectrum and high-efficiency bactericidal activity against bacteria, it was named "antibactetial peptides (ABP)", which translates to antibacterial peptides in Chinese, and its original meaning ...

Claims

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Application Information

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IPC IPC(8): A61K31/00C07K1/16C07K14/435
Inventor 张莉郭聪朱家裕
Owner 成都天创生物科技有限责任公司
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