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Method for preparing lactoferricin and method for applying lactoferricin in bacterial inhibition of foods

A technology of bovine lactoferrin peptide and fusion protein, which is applied in the preparation of lactoferrin peptide and its application in food bacteriostasis, and can solve the problem of recombinant expression bovine lactoferrin peptide and recombinant bovine lactoferrin peptide not involving Escherichia coli. problem, to achieve good inhibition effect, simple method and wide application range

Inactive Publication Date: 2012-06-20
SHANGHAI OCEAN UNIV
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  • Claims
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Problems solved by technology

[0004] None of the above-mentioned patents (applications) involves the use of Escherichia coli to recombinantly express bovine lactoferrin peptide and recombinant bovine lactoferrin peptide for antibacterial treatment of food, especially milk and aquatic products, so as to improve the food safety of food, especially raw seafood sex

Method used

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  • Method for preparing lactoferricin and method for applying lactoferricin in bacterial inhibition of foods

Examples

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Effect test

Embodiment 1

[0034] The preparation method of lactoferrin peptide:

[0035] (i) Activation of genetically engineered bacteria: use an inoculation loop to inoculate a loop of genetically engineered bacteria in 5 mL of LB liquid medium containing kanamycin sulfate after activation, then inoculate in the fermentation medium at a ratio of 1.5% (v / v) Medium fermentation, 7 hours after lactose induction;

[0036] Among them, the fermentation medium is: soluble starch 7g / L, beef extract 30g / L, sodium chloride 6.7g / L

[0037] (ii) Bacterial lysis: Add about 80 mL of lysis buffer to 20 g of induced cells, mix well and incubate overnight at 37°C, centrifuge to remove the supernatant (8000 r / min, 20 min), add an equal volume of inclusion body washing buffer, 37 After incubating at ℃ for 2-3 hours, centrifuge to remove the supernatant (8000r / min, 20min);

[0038] Wherein, the cell lysis buffer is: lysozyme (Lysozyme), its mass percentage is 0.02%, polyethanol octyl phenyl ether (Triton X-100), its v...

Embodiment 2

[0045] Put 25g of arctic shellfish inoculated with Vibrio parahaemolyticus into 225mL of bacteriostatic solution [bovine lactoferrin peptide isolated in Example 1: 2000μg / mL] for 30 minutes, store it at a low temperature of 4°C for 3 days, and measure it by the national standard method Vibrio parahaemolyticus and the total number of bacteria, and normal saline as a blank control. The results showed that compared with the control group, the content of Vibrio parahaemolyticus in arctic shellfish treated with antibacterial solution was reduced by 4.1×10 5 CFU / g.

Embodiment 3

[0047] Put 25g of arctic shellfish inoculated with Vibrio parahaemolyticus into 225mL of bacteriostatic solution [concentration of bovine lactoferrin peptide isolated in Example 1: 3000 μg / mL] for 30 minutes, then store at low temperature 4°C for 3 days, using the national standard method The total number of Vibrio parahaemolyticus and bacteria was determined, and normal saline was used as a blank control. The results showed that compared with the control group, the content of Vibrio parahaemolyticus in arctic shellfish treated with antibacterial solution was reduced by 5.1×10 5 CFU / g.

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Abstract

The invention relates to a method for preparing lactoferricin and a method for applying the lactoferricin in bacterial inhibition of foods. The method for preparing the lactoferricin comprises the following steps of: (i) activating genetic engineering bacteria; (ii) performing bacterial lysis; (iii) separating an inclusion body and performing lysis; (iv) extracting fusion protein and performing acid hydrolysis; and (v) separating by using a chromatographic column. The method is feasible, easy to operate and low in cost; and the lactoferricin prepared by the method can be used for bacterial inhibition of aquatic products. The method for applying the lactoferricin comprises the following steps of: cleaning an aquatic product, and soaking in a solution of the lactoferricin at normal temperature or at the temperature of between 0 and 4 DEG C for 0.5 to 10 hours.

Description

technical field [0001] The invention relates to a preparation and application method of a bacteriostatic agent, in particular to a preparation method of lactoferrin peptide and its application method in food bacteriostasis. Background technique [0002] Food safety is a major issue related to the national economy and the people's livelihood, and the development and application of all-natural green antibacterial agents is a frontier topic in China and the world. Lactoferrin (LF) is a naturally occurring iron-binding glycoprotein widely distributed in human and livestock milk, saliva, tears and other exocrine fluids. It is safe and harmless. Lactoferrin polypeptide (Lactoferricin, Lfcin) is a small peptide of LF hydrolyzed by acid pepsin in the digestive tract. A large number of studies have shown that Lfcin has broad-spectrum antibacterial, antiviral, and antifungal activities. Although they do not have iron ion binding sites, their bactericidal activity is higher than that ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/02C07K1/36C07K1/18C07K1/16C07K1/12A23L3/3526C12R1/19
Inventor 刘承初巴特邓强鲁建章李家乐
Owner SHANGHAI OCEAN UNIV
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