阅读说明：本技术 红曲抗菌肽hqP3及其应用 (Red yeast antibacterial peptide hqP3 and application thereof ) 是由 陈翠贤 王思捷 于 2019-09-14 设计创作，主要内容包括：本发明提供了一种来源于红曲的新型抗菌多肽hqP3,其氨基酸序列如SEQ ID NO：3所示,其基因编码序列如SEQ ID NO：7所示；多肽hqP3对于大肠杆菌和铜绿假单胞菌均具有良好的杀菌作用,最小杀菌浓度分别低至0.16μg/ml和0.08μg/ml,并且不产生耐药菌；此外多肽hqP3对于同时耐哌拉西林、环丙沙星和头孢他啶的铜绿假单胞菌多重耐药菌以及同时耐哌拉西林、甲氧苄啶、环丙沙星和庆大霉素的大肠杆菌多重耐药菌均具有杀灭作用,有望做为抗生素替代品,具有广阔的应用前景。(The invention provides a novel antibacterial polypeptide hqP3 from red yeast rice, the amino acid sequence of which is shown as SEQ ID NO: 3, the gene coding sequence is shown as SEQ ID NO: 7 is shown in the specification; the polypeptide hqP3 has good bactericidal effect on escherichia coli and pseudomonas aeruginosa, the minimum bactericidal concentration is respectively as low as 0.16 mug/ml and 0.08 mug/ml, and drug-resistant bacteria are not generated; in addition, the polypeptide hqP3 has a killing effect on pseudomonas aeruginosa multi-drug resistant bacteria simultaneously resistant to piperacillin, ciprofloxacin and ceftazidime and escherichia coli multi-drug resistant bacteria simultaneously resistant to piperacillin, trimethoprim, ciprofloxacin and gentamicin, is expected to be used as an antibiotic substitute, and has a wide application prospect.)

1. A monascus antimicrobial peptide hqP3 is characterized in that the amino acid sequence of antimicrobial peptide hqP3 is shown in SEQ ID NO: 3, the gene coding sequence is shown as SEQ ID NO: shown at 7.

2. The antibacterial peptide hqP3 of claim 1, wherein the antibacterial peptide hqP3 has good bactericidal effect on both escherichia coli and pseudomonas aeruginosa, has minimum bactericidal concentrations as low as 0.16 μ g/ml and 0.08 μ g/ml respectively, and does not produce drug-resistant bacteria.

3. The antibacterial peptide hqP3 of claim 1, wherein the antibacterial peptide hqP3 has killing effect on pseudomonas aeruginosa multi-drug resistant bacteria simultaneously resistant to piperacillin, ciprofloxacin and ceftazidime and escherichia coli multi-drug resistant bacteria simultaneously resistant to piperacillin, trimethoprim, ciprofloxacin and gentamicin.

4. The use of the antimicrobial peptide hqP3 of claim 1, 2 or 3 in the fields of clinical therapeutics, food preservation and preservation, and animal feed.

Technical Field

The invention relates to the field of molecular biology, in particular to an antibacterial peptide from red yeast rice and application thereof.

Background

Chinese yellow wine is one of three ancient wines in the world and is a special national product in China. It is made up by using grain as raw material and making it by using yeast. It is well received by the masses because of its rich nutrition, low alcohol content and unique flavor, taste and efficacy. The Fujian red rice yellow wine is one of Chinese yellow wine with unique color, and is brewed with glutinous rice and rice as main material and through mixing red rice and white yeast, and is famous for red color, mellow taste and strong fragrance. The red yeast yellow wine not only has rich nutrient components, but also has distinctive physiological effects due to the addition of red yeast for fermentation. The yellow wine brewing can be divided into two stages of yeast making and brewing, the traditional brewing process is a group micro-fermentation process, and needs the participation of various microorganisms, and the yeast can provide the microorganisms needed in the brewing process, so the yeast is indispensable to the brewing. For a long time, the traditional distiller's yeast including red yeast rice is completely prepared by a natural microorganism inoculation mode, and the sources of microorganisms mainly include mother yeast, brewing raw materials, water, utensils, yeast preparation places and the like. These microbial sources contain both brewing-relevant microorganisms and a large number of other microorganisms, and therefore, a corresponding ecological mechanism must exist in the process of forming the distiller's yeast, so that the brewing-relevant microorganisms have a competitive advantage to become the main flora and maintain the stability of the system.

Monascus is currently known to be the major microorganism in monascus, growing relatively slowly, but having unique metabolic properties. Research shows that monascus can produce rich secondary metabolites, mainly comprises monascus pigment, monacolin K, r-aminobutyric acid, and various antioxidant components such as Dimerumic acid, acetylcholine, ergosterol, citrinin and the like, the metabolites are important components of unique quality and physiological effect of monascus yellow wine, and monascus is named due to the monascus pigment produced by monascus. Besides the macroscopic pigments, monascus can also produce various antibacterial substances as the main microorganism in monascus. The ancient times in China have traditions of preserving food by using red yeast rice, for example, in the 'astronomy opening article', records about preserving flesh color for 10 days without rotting by using red yeast rice in hot summer are recorded, which indicates that the red yeast rice has bacteriostatic and bactericidal effects. Modern researches show that monascus in red yeast can generate antibacterial substances such as citrinin and the like, and monascus pigment, orange pigment and other pigments generated by the monascus also have a certain antibacterial function. Therefore, the red yeast rice contains abundant antibacterial substances inevitably, and can resist the breeding of harmful microorganisms and other mixed bacteria so as to generate and maintain the unique microbial ecological stability. In view of the fact that red yeast rice is a source of potential antibacterial substances which may contain harmful microorganisms, the applicant of the present invention has screened polypeptides having good specific bactericidal properties. The antibacterial peptide is used as a novel antibiotic, is not easily resistant to the antibiotic by microorganisms, has wide application prospect, and needs to be continuously developed to be used in the fields of clinical treatment medicines, food preservation and preservation, animal feed and the like.

Disclosure of Invention

The invention provides an antibacterial polypeptide hqP3 from red yeast rice for the first time, discloses a gene coding sequence thereof, explores an antibacterial spectrum thereof, determines the corresponding minimum bactericidal concentration, and also explores the drug resistance of target bacteria to the polypeptide hqP3 and the killing effect of the polypeptide hqP3 to drug-resistant target bacteria.

The technical scheme adopted by the invention for solving the technical problems is as follows:

The invention provides a novel antibacterial polypeptide hqP3 from red yeast rice, the amino acid sequence of which is shown as SEQ ID NO: 3, the gene coding sequence is shown as SEQ ID NO: 7 is shown in the specification; the polypeptide hqP3 has good bactericidal effect on escherichia coli and pseudomonas aeruginosa, the minimum bactericidal concentration is respectively as low as 0.16 mug/ml and 0.08 mug/ml, and drug-resistant bacteria are not generated; in addition, the polypeptide hqP3 has a killing effect on pseudomonas aeruginosa multi-drug resistant bacteria simultaneously resistant to piperacillin, ciprofloxacin and ceftazidime and escherichia coli multi-drug resistant bacteria simultaneously resistant to piperacillin, trimethoprim, ciprofloxacin and gentamicin.

The novel antibacterial polypeptide hqP3 derived from red yeast rice provided by the invention has strong bactericidal action on escherichia coli and pseudomonas aeruginosa, does not generate drug-resistant bacteria, can kill main drug-resistant bacteria of escherichia coli and pseudomonas aeruginosa, is expected to be an antibiotic substitute, and has wide application prospect.

Drawings

FIG. 1 is a diagram showing the relationship between the absorbance and the antibacterial efficacy of Sm monascus protein isolate.

FIG. 2 is a graph showing experimental results of air contact antimicrobial efficacy of polypeptide hqP 1.

FIG. 3 is a graph of experimental results of air contact antimicrobial efficacy of polypeptide hqP 2.

FIG. 4 is a graph of experimental results of air contact antimicrobial efficacy of polypeptide hqP 3.

FIG. 5 is a graph of experimental results of air contact antimicrobial efficacy of polypeptide hqP 4.

Fig. 6 is a graph showing experimental results of air-contact antibacterial efficacy of the blank control group.

Detailed Description

The invention is further illustrated below with reference to specific examples.