Construction and application of recombinant protein based on lactobacillus LPxTG motif

A technology of recombinant protein and lactic acid bacteria, applied in the field of lactic acid bacteria, to achieve the effect of good tolerance

Active Publication Date: 2022-03-11
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are few studies on the combination of lactic acid bacteria LPxTG surface structural proteins and functional peptides in the prior art, so it can be further explored

Method used

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  • Construction and application of recombinant protein based on lactobacillus LPxTG motif
  • Construction and application of recombinant protein based on lactobacillus LPxTG motif
  • Construction and application of recombinant protein based on lactobacillus LPxTG motif

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1: Construction method based on lactic acid bacteria LPxTG motif recombinant protein

[0054] 1.1 obtain the gene fragment of LPxTG-functional peptide (step S1)

[0055] According to the gene sequence of the surface structural protein containing the LPxTG sequence in Lactobacillus reuteri published on NCBI:

[0056] (As shown in SEQ ID NO.1), use CE Design to design primers to obtain primers F1 and R1, and increase the sequence of functional peptides in primer F1 (in this embodiment, functional peptides are antioxidant peptides, and its amino acid sequence is shown in the table 1. After the functional peptide was synthesized by Shanghai Sangon Biological Co., Ltd., corresponding to the F1' of the primer, see Table 1, and the designed primer sequence was sent to Shanghai Sangon Biological Co., Ltd. for synthesis. The whole genome of Lactobacillus reuteri SH23 was extracted with the Easy Pure BacteriaGenomic DNA Kit, and the DNA concentration was verified with...

Embodiment 2

[0070] Example 2: Comparative study on the functional properties of LPxTG-functional peptides and functional peptides

[0071] In Example 2, in order to reflect the antioxidant capacity of the functional peptide, the functional peptide was named as an antioxidant peptide. That is, the LPxTG-functional peptide in Example 1 is the LPxTG-antioxidative peptide in Example 2; the functional peptide in Example 1 is the antioxidant peptide in Example 2.

[0072] In order to compare the scavenging activities of LPxTG-antioxidant peptides and antioxidant peptides on hydroxyl radicals, ABTS radicals and DPPH radicals, the following experiments were carried out.

[0073] 2.1 Hydroxyl radical scavenging activity

[0074] Divided into 8 groups. Among them, group 1 has only antioxidant peptide (i.e. peptide), the concentration is 5 μg / ml, that is, dissolving 5 μg antioxidant peptide per milliliter of distilled water; group 2 is only LPxTG-antioxidant peptide (i.e. LPxTG-peptide), the conce...

Embodiment 3

[0088] Example 3: Test method for gastrointestinal tolerance of LPxTG-functional peptides

[0089] In Example 3, in order to reflect the antioxidant capacity of the functional peptide, the functional peptide was named antioxidant peptide. That is, the LPxTG-functional peptide in Example 1 is the LPxTG-antioxidative peptide in Example 3; the functional peptide in Example 1 is the antioxidant peptide in Example 3.

[0090] In order to test the gastrointestinal tolerability properties of LPxTG-antioxidant peptides, the following experiments were performed. Artificial gastric juice (hereinafter referred to as gastric juice for short) and artificial intestinal juice (hereinafter referred to as intestinal juice for short) were prepared respectively. The preparation method of artificial gastric juice is to add 0.5g NaCl, 0.3g pepsin and dilute to 100ML with ultrapure water, pH=2. The preparation method of artificial intestinal juice is to add 0.5g NaCl, 0.5g ox bile salt, 0.3g trypsi...

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Abstract

The invention discloses a recombinant protein based on a lactic acid bacteria LPxTG motif. The recombinant protein (LPxTG-functional peptide) has better tolerance to the gastrointestinal tract environment, so that the functional peptide can be protected, and the damage of gastric juice and intestinal juice to the functional peptide in the oral administration process is reduced. The invention also discloses a construction method of the recombinant protein based on the lactic acid bacteria LPxTG motif, an application of the construction method of the recombinant protein based on the lactic acid bacteria LPxTG motif in improving the oxidation resistance of functional peptides, and an application of the recombinant protein based on the lactic acid bacteria LPxTG motif in oral preparations.

Description

technical field [0001] The invention relates to the technical field of lactic acid bacteria, in particular to the construction and application of a recombinant protein based on lactic acid bacteria LPxTG motif. Background technique [0002] As an important component of living organisms, protein is an important material basis for living organisms. Many studies have found that the peptides after protein degradation have many physiological and functional properties, such as anti-oxidation, anti-hypertension, anti-bacterial, anti-cancer and other activities, which are referred to as functional peptides for short. Among them, functional peptides with antioxidant properties are called for antioxidant peptides. Compared with proteins, functional peptides not only have the same amino acid composition as homologous proteins, but also are easier to be absorbed by the intestines than proteins. More studies have shown that the binding of functional peptides to intestinal receptors may ...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/70A61K38/16A61K9/00A61P39/06
CPCC07K14/335C07K7/06C12N15/70A61K9/0053A61P39/06C07K2319/00A61K38/00Y02A50/30
Inventor 吴振卢淑怡潘道东劳立峰孙杨赢蓝航镇
Owner NINGBO UNIV
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