Solid fermentation preparation method of antibacterial peptide and application

A technology of solid fermentation and antimicrobial peptides, applied in the application of pharmaceuticals and preservative products, and in the field of preparation of feed additives, can solve problems that affect the correct folding of products and reduce product activity, and achieve easy promotion, simple production process, and easy cultivation and reproduction Effect

Active Publication Date: 2009-03-11
SHENZHEN SUNSMILE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this involves a series of complex molecular biological operations, and there is great uncertainty in cutting the tandem expression product in

Method used

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  • Solid fermentation preparation method of antibacterial peptide and application
  • Solid fermentation preparation method of antibacterial peptide and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The preparation of antimicrobial peptide (Clavanin H) operates as follows:

[0018] (1) Bacterial strain activation: The fungal strains of this embodiment are selected from common fungal strains in the art, preferably Saccharomyces (Saccharomyces) fungi are strains, called baker's yeast (Saccharomyces cerevisiae), preservation number CICC1795, purchased from Guangdong Microbiology graduate School. Baker's yeast, also known as Saccharomyces cerevisiae, contains high-quality protein, dietary fiber, vitamin B complex and organic chromium, zinc, selenium, iron, calcium and other elements that meet the needs of the body. It is a pure natural, pollution-free, biological Source of healthy nutrition. At the same time, because yeast has many identical structures with animal and plant cells that are also eukaryotes, and is similar to prokaryotes for easy cultivation, it is used as a eukaryotic model organism in modern molecular and cell biology, and is an ideal Genetic engineer...

Embodiment 2

[0024] The difference between this embodiment and embodiment 1 is that its solid fermentation step also includes:

[0025] Take the solid fermentation culture based on 121°C, 20min sterilization, and after cooling, insert the third-grade seeds into it according to the inoculation amount of 10% by mass ratio, stir and mix well, keep the humidity at 60%, and the pH at the natural value, at 28°C Cultivate for 1 day to obtain the fourth-grade seeds; take the solid fermentation medium, sterilize it at 121°C for 20 minutes, and cool it down, insert the fourth-grade seeds into it according to the inoculation amount of 10% by mass, stir and mix well, and keep the humidity The pH is 60%, the pH is the natural value, and cultured at 28° C. for 1 day to obtain the fifth-order seeds, that is, the antimicrobial peptide product. Certainly, in this step, the mass ratio of the inoculum amount is all feasible within the range of 5-15%.

[0026] The antimicrobial peptides produced can be direc...

Embodiment 3

[0031] Antimicrobial peptides inhibit the pathogenic bacteria of cow mastitis. In the test, the milk of dairy cows with obvious symptoms of mastitis such as inflamed and swollen breasts, red and warm skin, poor milk discharge, thin yellowish mixed with flocs and curds was selected. In the experimental group, 5 ml of the above-mentioned dairy cow milk was mixed with the antibacterial peptide obtained by the method of the above-mentioned embodiment at a concentration of 0.5 mg / ml for 30 minutes, and in the control group, 5 ml of the above-mentioned cow milk was mixed with sterilized physiological saline for 30 minutes. The above mixed solutions were spread on LB solid medium respectively, and the growth of the pathogenic bacteria of mastitis in dairy cows was observed. The experimental results showed that the milk of cows added with antimicrobial peptides was spread on the medium, and the mastitis pathogenic bacteria did not grow at all; while the milk of cows without antimicrob...

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Abstract

The invention provides a method for preparing antibacterial peptide, which comprises the following steps: firstly, preparation of primary seeds; and secondly, solid fermentation. Because the method adopts the means of solid fermentation to prepare the antibacterial peptide, large-scale equipment investment is not required; the manufacturing technique is simple and convenient; acclimated engineering bacteria are easy to culture, propagate and produce; fermented products do not require separation and purification and can be directly applied; and the method is easy to promote. Moreover, the antibacterial peptide produced by the method has antibacterial activity with broad spectrum, and not only has strong killing effect on harmful bacteria such as golden yellow staphylococcus, mildew and so on and fungus but also has killing effect on viruses.

Description

technical field [0001] The invention relates to a preparation method of an antibacterial peptide and the application of the antibacterial peptide product in the preparation of feed additives, medicines and preservatives. Background technique [0002] Antimicrobial peptides are small molecular polypeptides with biological activity induced by organisms, generally composed of 20-60 amino acids, and the molecular weight is about 2000-7000 Daltons. Since the Swedish scientist Boman et al. isolated the first antibacterial peptide cecropin (Cecropin) from silkworm chrysalis in 1975, people have been widely used in insects, amphibians, aquatic animals, mammals including humans, even plants and bacteria. At least 900 antimicrobial peptides have been found in the biological spectrum. This kind of short peptide not only has a broad-spectrum bactericidal effect on bacteria and fungi, but also has a killing effect on certain viruses, protozoa and tumor cells. Clinical trials have shown...

Claims

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Application Information

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IPC IPC(8): C12P21/00A23K1/16A61K38/10A61K38/16A61P31/00A23L3/3526C12R1/85A23K20/147
Inventor 苏向东彭永鹤李永新黄毓茂皮灿辉
Owner SHENZHEN SUNSMILE BIOTECH
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