阅读说明：本技术 对多种病原菌具有抑菌活性的细菌素 (Bacteriocin with bacteriostatic activity against various pathogenic bacteria ) 是由 信丙越 李峰 刘舒 曾化伟 曾昕 王光利 张标 徐大勇 于 2020-05-21 设计创作，主要内容包括：本发明公开了一种对多种病原菌具有抑菌活性的细菌素bacicyclicin XIN-1,是一种新型环状细菌素,分子量为5848.2032 Da,其前体肽的核苷酸序列如SEQ ID NO：1所示,氨基酸序列如SEQ ID NO：2所示。所述细菌素bacicyclicin XIN-1产自相山芽胞杆菌XIN-1,对多种病原菌如蜡样芽胞杆菌、单核细胞增生李斯特菌、金黄色葡萄球菌等具有高效抑菌活性。(The invention discloses a bacteriocin XIN-1 with bacteriostatic activity to various pathogenic bacteria, which is a novel annular bacteriocin with molecular weight of 5848.2032Da, and the nucleotide sequence of a precursor peptide is shown as SEQ ID NO:1, and the amino acid sequence is shown as SEQ ID NO:2, respectively. The bacteriocin bacicicin XIN-1 is produced from Bacillus faciopneumoniae XIN-1, and has high-efficiency antibacterial activity on various pathogenic bacteria such as Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus and the like.)

1. A bacteriocin XIN-1 having bacteriostatic activity against a plurality of pathogenic bacteria, characterized in that:

the bacteriocin bacicicin XIN-1 is a cyclic bacteriocin with molecular weight of 5848.2032 Da.

2. The bacteriocin bacicicin XIN-1 according to claim 1, characterized in that:

the nucleotide sequence of the precursor peptide of the bacteriocin bacicicin XIN-1 is shown as a sequence table SEQ ID NO. 1.

3. The bacteriocin bacicicin XIN-1 according to claim 1, characterized in that:

the amino acid sequence of the bacteriocin bacicicin XIN-1 is shown in a sequence table SEQ ID NO. 2.

4. The bacteriocin bacicicin XIN-1 according to claim 1, 2 or 3, characterized in that:

the bacteriocin bacicicin XIN-1 is produced from Bacillus faciaensis XIN-1.

5. The bacteriocin bacicicin XIN-1 according to claim 4, wherein:

the classification of the Bacillus phaericus XIN-1 is named as: bacillus xiangshan XIN-1, deposited unit: china Center for Type Culture Collection (CCTCC), address: wuhan university, preservation date: year 2020, 4/2, accession number: CCTCC NO: m2020064.

Technical Field

The invention belongs to the field of microorganisms, and particularly relates to a bacteriocin XIN-1 with antibacterial activity on various pathogenic bacteria.

Background

With the emphasis on food safety and self health of people, the traditional preservation technology such as chemical preservatives can cause adverse effects on blood pressure, heart, kidney and the like, and even some problems of cancer inducement, teratogenicity and the like can not meet the needs of people, so that the search for natural and safe food preservatives becomes a research hotspot in the food industry.

Bacteriocins are a class of proteins or polypeptides with antibacterial activity produced by bacteria via the ribosome synthesis mechanism. Bacteriocins themselves have many advantageous properties: high bacteriostatic activity in vivo and in vitro; is nontoxic to human body or animals, and is easily degraded by some protease in animal digestive tract; has good selective sterilization effect; the biological modification is easy, and different requirements of people are met; has thermal stability and acid and alkali resistance, and is convenient for storage; tasteless in nature and does not affect the mouthfeel, taste, etc. of foods, feeds when used (Cotter et al, 2013). Therefore, the bacteriocin has wide application value and wide application prospect in the industries of food, feed, medicine and the like.

Bacillus is a group of gram-positive bacteria that are widely distributed in various ecoenvironments and can form spores. Research shows that Bacillus (Bacillus) can produce various bacteriocins, such as bacteriocin CD (human bacterial CD) produced by a strain of Bacillus thuringiensis separated from human stool samples can specifically inhibit clostridium difficile, and can not cause imbalance of intestinal flora in a human colon model, so that Bacillus (Bacillus) is a novel medicament for resisting clostridium difficile infection in the future (read et al, 2010). In addition, 3 novel lantibiotins, namely TiCin A1, TiCin A3 and TiCin A4, are identified in Bacillus thuringiensis BMB3201 by the subject group, have high-efficiency bacteriostatic activity on pathogenic foodborne bacteria Bacillus cereus and Listeria monocytogenes, have extremely high heat resistance and acid-base characteristics, and have application potential (Xin et al, 2015) as a novel food preservative.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide the bacteriocin XIN-1 which has bacteriostatic activity on various pathogenic bacteria. The bacteriocin bacicicin XIN-1 provided by the invention has high-efficiency antibacterial activity on various pathogenic bacteria.

The bacteriocin XIN-1 with bacteriostatic activity to various pathogenic bacteria is a novel cyclic bacteriocin with molecular weight of 5848.2032Da (measured by using a single isotope).

The nucleotide sequence of the precursor peptide of bacteriocin XIN-1 is shown in a sequence table SEQ ID NO. 1, and the amino acid sequence is shown in a sequence table SEQ ID NO. 2.

SEQ ID NO. 1 of the sequence Listing is a nucleotide sequence of a precursor peptide of bacicicin XIN-1, and consists of 195 nucleotides in total.

SEQ ID NO 2 of the sequence Listing is the amino acid sequence of the precursor peptide of bacicicin XIN-1, and codes 64 proteins in total.

The bacteriocin bacicicin XIN-1 is produced from Bacillus faciopneumoniae XIN-1, and the classification and designation of the Bacillus faciopneumoniae XIN-1 are as follows: bacillus xiangshan XIN-1, deposited unit: china Center for Type Culture Collection (CCTCC), address: wuhan university, preservation date: year 2020, 4/2, accession number: CCTCC NO: m2020064.

The invention determines that the Bacillus licheniformis XIN-1(Bacillus xiangshanXIN-1) strain can generate substances with bacteriostatic activity on various pathogenic bacteria (such as Bacillus cereus ATCC14579, Listeria monocytogenes ATCC19115 and staphylococcus aureus ATCC29523) through bacteriostatic activity screening. The bacteriostatic active substance produced by the extracted Bacillus facioplicans XIN-1 strain is a novel cyclic bacteriocin, and the bacteriocin substance is named as bacicicin XIN-1. baciciclin XIN-1 has antibacterial activity against various gram-positive pathogenic bacteria (such as Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, etc.).

Drawings

FIG. 1: and (3) detecting the bacteriostatic activity of fermentation supernatants of different growth periods of the Bacillus phaericus XIN-1 strain on Bacillus cereus ATCC14579, Listeria monocytogenes ATCC19115 and Staphylococcus aureus ATCC 29523.

FIG. 2: HPLC analysis and mass spectrometry analysis results of bacteriocin-bacicicin XIN-1 produced by Bacillus phaeodorus XIN-1 strain.

FIG. 3: comparative analysis of synthetic Gene Cluster and precursor peptide sequence of baciciclin XIN-1. Description of reference numerals: FIG. 3A is a synthetic gene cluster of baciciclin XIN-1, and FIG. 3B is a functional analysis of each gene in the synthetic gene cluster of baciciclin XIN-1; FIG. 3C is an alignment analysis of the sequence of baciciclin XIN-1 precursor peptide with the reported cyclic bacteriocin precursor peptide.

FIG. 4: structure of baciciclin XIN-1.

Detailed Description