50 results about "Phage lysis" patented technology

The invention provides a kit for rapidly detecting MRSA and drug-resistant gene nucleic acid POCT of the MRSA based on a phage lyase mutant. The kit comprises nucleic acid extraction and purification and triple PCR rapid amplification detection reagents. The staphylococcus aureus bacteriophage lyase mutant nucleic acid release liquid containing the amino acid sequence as shown in SEQ ID NO: 1 is adopted, wall breaking can be rapidly conducted on staphylococcus aureus, and nucleic acid is released. The effect of the nucleic acid release liquid acting for 5 minutes is equivalent to or even better than that of the lysozyme acting for 60 minutes; according to the present invention, with the combination of the MRSA and drug-resistant gene triple PCR rapid amplification detection reagent, the total detection time is only 35 min, the time is saved by 55 min or more than 55 min compared with the lysozyme method, and the MRSA and drug-resistant gene nucleic acid POCT rapid detection can be achieved;

The invention provides engineering lyase Art-15 with broad-spectrum antibacterial activity, antibacterial peptide NZ2114 and bacteriophage lyase PlySs2 are subjected to functional fusion through an optimally designed flexible linker, and an efficient bactericidal effect on Staphylococcus aureus (Staphylococcus aureus) and Streptococcus spp.) is achieved. The engineering lyase Art-15 has the advantages that the engineering lyase Art-15 has broad-spectrum antibacterial activity, the antibacterial peptide NZ2114 and the bacteriophage lyase PlySs2 are subjected to functional fusion through the optimally designed flexible linker, and the engineering lyase Art-15 has broad-spectrum antibacterial activity; the lyase Art-15 overcomes the defects of narrow host spectrum and insufficient splitting activity of the lyase through a synergistic destruction mechanism of targeting bacterial cell wall peptidoglycan. Experiments show that the minimal inhibitory concentration (MIC) of the lyase Art-15 on staphylococcus aureus in vitro is lower than 2 mu g / mL, the minimal inhibitory concentration of the lyase Art-15 on streptococcus agalactiae is lower than 1 mu g / mL, the minimal inhibitory concentration of the lyase Art-15 on streptococcus dysgalactiae is lower than 2 mu g / mL, and the sterilization rate of the lyase Art-15 can reach 97% or above within 30 minutes by 2 times of MIC. The lyase Art-15 disclosed by the invention has important application value in the fields of drug-resistant bacterium infection treatment, medical instrument disinfection and livestock breeding.

The invention discloses a bacteriophage lyase and application thereof, and particularly relates to a bacteriophage lyase with an amino acid sequence as shown in SEQ INNO.2. The bacteriophage lyase can inhibit the growth of escherichia coli, staphylococcus and salmonella, can be used as an antibacterial substance in splitting gram-negative bacteria and can be used for preparing drugs for resisting gram-negative bacteria. The lyase can be used independently or compounded with other substances, the use concentration of antibiotics can be reduced by combining the lyase with the antibiotics, toxic and side effects (such as renal toxicity of polymyxin) of drugs are reduced, and meanwhile, generation of bacterial drug resistance is delayed. After the lyase and the chitosan are combined for use, remarkable antibacterial activity is generated, the antibacterial effect can be achieved without pre-treatment on bacteria, and the practicability is improved.

The invention belongs to the technical field of biology, and particularly relates to perforin derived from staphylococcus warneri mild bacteriophage and application. Perforin has a nucleotide sequence as shown in SEQ ID NO: 1. The perforin is a key gene for bacteriophage splitting growth and plays an important role in the splitting process of host bacteria. Therefore, the research aims to verify whether the perforin of which the coding site is positioned on a staphylococcus warneri mild bacteriophage genome can be used as a cell lysis gene and can be used as a potential antibacterial substance or not, and a theoretical basis is provided for prevention and treatment of gram-negative pathogenic bacteria.

The utility model discloses a bacteriophage separating device for culturing bacteriophages, which relates to the technical field of bacteriophage culturing and separating devices and comprises a base, vertical columns fixedly mounted at four corners of the bottom of the base, a container arranged at the upper end of the base, a feeding pipe communicated and fixedly mounted at the upper end of the container, and a discharging pipe communicated and fixedly mounted at the bottom end of the container. A sealing plug is arranged at the discharge pipe, and one end of the sealing plug is in threaded connection with the interior of the discharge pipe and can block the sealing plug. According to the bacteriophage separation device for culturing the bacteriophage, the annular plate is driven to rotate by rotating the connecting piece, the annular plate is divided into a plurality of strip shapes by the strip-shaped grooves, and the annular plate can block the filter groove, so that the annular container is separated from the container, and a bacteriophage lysis solution is prevented from flowing back into the container from the filter groove. The backflow phenomenon is avoided, so that the phage can keep high purity in the separation process, and a high-quality phage sample is provided for subsequent experiments and applications.

The invention relates to the technical field of biological medicine, in particular to a composition for blocking formation of phage treatment tolerance and application of the composition. The composition comprises a folium ilicis purpurea ethyl acetate extraction active component and a clostridium perfringens specific bacteriophage, the folium ilicis purpurea ethyl acetate extraction active component is prepared from the following components in percentage by mass: 8 to 15 percent of protocatechuic acid and 5 to 10 percent of ethyl gallate, and the mass ratio of the protocatechuic acid to the ethyl gallate is (1.5 to 1) to (2.5 to 1); the clostridium perfringens specific bacteriophage is a virulent bacteriophage capable of specifically cracking a clostridium perfringens NetB toxin positive strain. According to the invention, the shallow thought that the existing compounding technology only pursues synergistic sterilization is subverted, and a deep synergistic mode of bacteriophage splitting attack and folium ilicis purpurea component disintegration bacterial defense program is proposed and verified for the first time. And the formation of tolerance is fundamentally blocked, so that a more thorough and more lasting treatment effect is realized, and the comprehensive production performance of animals is accidentally and greatly improved on the basis of the treatment effect.

The invention discloses application of bacteriophage lyase LysFEC14 in preparation of a composition for resisting gram-negative bacteria. The amino acid sequence of the bacteriophage lyase LysFEC14 is as shown in SEQ ID NO.01. The invention further discloses a preparation method of the bacteriophage lyase LysFEC14. The antibacterial composition disclosed by the invention shows high-efficiency cracking activity on various escherichia coli standard strains, still has a remarkable antibacterial effect at low concentration, and provides a green and efficient novel antibacterial composition for preventing and treating gram-negative bacterium infection, particularly drug-resistant escherichia coli infection.

This invention belongs to the field of biotechnology, specifically relating to a perforin derived from *Staphylococcus warwickiana* temperate bacteriophage and its applications. The perforin has the nucleotide sequence shown in SEQ ID NO:1. This perforin is a key gene for phage lysis and growth, playing a crucial role in the lysis of host bacteria. Therefore, this study aims to verify whether the perforin, whose coding site is located on the genome of *Staphylococcus warwickiana* temperate bacteriophage, as a cell lysis gene, can serve as a potential antibacterial substance, providing a theoretical basis for the prevention and control of Gram-negative pathogens.

The invention discloses recombinant bacteriophage lyase and application thereof. The amino acid sequence of the recombinant bacteriophage lyase E8-3 is as shown in SEQ ID NO: 43, and an in-vitro minimum inhibitory concentration experiment shows that the recombinant bacteriophage lyase E8-3 has a bacteriostatic effect on helicobacter pylori and acinetobacter baumannii, and particularly has a wide application prospect in the development of anti-helicobacter pylori drugs.

The invention discloses a fusion protein containing bacteriophage lyase and LPS (lipopolysaccharide) binding peptide, and further modifications including amino acid mutation, combination mode change and chemical modification are carried out on the fusion protein. The bactericidal activity of the bacteriophage lyase is improved, and meanwhile, the in-vivo stability is improved. The fusion protein provided by the invention is suitable for preventing and treating diseases caused by bacterial infection, especially gram-negative bacterial infection.

The invention provides application of aeromonas hydrophila bacteriophage lyase Lys56 in preparation of a bacteriostatic agent, and belongs to the technical field of biology. The aeromonas hydrophila bacteriophage lyase Lys56 disclosed by the invention has an inhibiting effect on certain gram-negative bacteria and gram-positive bacteria, and has no splitting effect on certain probiotics. The Lys56 is not resistant to high temperature and has high acid-base tolerance, the inhibition effect of the Lys56 can be enhanced by low-concentration divalent metal ions, the inhibition effect of the Lys56 can be enhanced by combined use of 1-10 mM of ethylenediamine tetraacetic acid (EDTA) and the Lys56, and the Lys56 can effectively inhibit and remove biofilms of host bacteria. In an artificial pollution model of the fish blocks, the antibacterial effect of the Lys56 is in direct proportion to the concentration of the Lys56. Lys56 has a good sterilization effect and has a good application prospect.

The invention discloses application of an antibacterial agent and bacteriophage lyase. The bacteriophage lyase has a remarkable splitting effect on gram-negative bacteria, so that the bacteriophage lyase can be used as a broad-spectrum antibacterial protein. For example, the antibacterial effect on gram-negative bacteria such as escherichia coli, pseudomonas aeruginosa and klebsiella pneumoniae is remarkable. The antibacterial protein has relatively high temperature tolerance and pH tolerance, can replace antibiotics, and is applied to prevention and treatment of bacterial diseases as an external medicine or an internal medicine and sterilization of medical instruments and medical places; and the product can also be used as an additive to be applied to the fields of feed processing, food industry, animal husbandry, beer industry, aquaculture, cosmetic production, fruit preservation and the like.

The invention relates to the technical field of biological medicines, discloses a broad-spectrum bacteriophage composition for preventing and controlling swine atrophic rhinitis and application of the broad-spectrum bacteriophage composition, and particularly relates to a pasteurella bacteriophage SD-PM1 and a bordetella bronchiseptica bacteriophage SD-Bb1, and the preservation numbers of the pasteurella bacteriophage SD-PM1 and the bordetella bronchiseptica bacteriophage SD-Bb1 are CCTCC NO: M20232599 and CCTCC NO: M20232589 respectively. The bacteriophage and the composition thereof have a strong broad-spectrum lysis effect on porcine pasteurella and porcine bordetella bronchiseptica, and the bacteriophage can be independently compounded with cocktail or compounded with cocktail, and can be used for preparing medicines for preventing and treating porcine respiratory diseases caused by independent infection or mixed infection of pasteurella and bordetella bronchiseptica. The traditional Chinese medicine composition especially has a remarkable effect on atrophic rhinitis of pigs. In addition, the bacteriophage provided by the invention is a strong bacteriophage, has a wide lysis spectrum, has good biological characteristics of acid resistance and heat resistance, and can also be applied to various actual scenes.

The invention relates to the technical field of biology, in particular to a broad-spectrum bacteriophage lyase mutant, an extraction and purification reagent and a kit. The invention provides a broad-spectrum bacteriophage lyase mutant, an extraction and purification reagent and a kit. A broad-spectrum bacteriophage lyase mutant nucleic acid release liquid containing protein with an amino acid sequence as shown in SEQ ID NO: 1 is adopted; the compound can efficiently act on two important clinical drug-resistant bacteria, namely methicillin-resistant staphylococcus aureus MRSA and vancomycin-resistant enterococcus VRE, at the same time. Through unique molecular design and preparation process optimization, the lyase shows excellent synchronous wall breaking capacity, cell wall destruction can be completed in an extremely short time, and the nucleic acid release efficiency is remarkably superior to that of a traditional method.

The invention provides bacteriophage lyase or a mutant thereof. The bacteriophage lyase comprises any one of the following items: (1) the bacteriophage lyase has an amino acid sequence as shown in SEQ ID NO.1 or has at least 80% identity with the amino acid sequence as shown in SEQ ID NO.1; (2) mutation in the amino acid sequence shown in (1) and in a polypeptide structural domain, a cell wall binding structural domain and / or an amidase structural domain with cysteine and histidine-dependent aminopeptidase (CHAP) activity; the temperature stability of the bacteriophage lyase and the cracking activity of the bacteriophage lyase on staphylococcus aureus are improved, dependence of the bacteriophage lyase on refrigeration conditions is reduced, side effects caused by antibiotics can be effectively avoided, and the bacteriophage lyase can be used for preventing or treating diseases caused by the staphylococcus aureus and has good application prospects. The growth of the staphylococcus aureus in the environment, production facilities, food or animal feed is inhibited, and biological membranes generated by the staphylococcus aureus are prevented and treated.

This invention discloses a strain of Salmonella choleraesuis phage SCP512 and its applications, belonging to the field of microbial technology. Salmonella choleraesuis phage SCP512 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 47021, dated February 4, 2026. It belongs to the family Myocaudae within the class Caudata. This phage SCP512 possesses multivalent lytic characteristics, excellent biological properties and environmental adaptability, and can disrupt pathogen biofilm formation. It addresses the technical problems of existing phages, such as limited lytic spectrum, insufficient coverage of common foodborne pathogens, weak biofilm removal ability, and limited application potential in practical food matrices.

The invention relates to the field of biology, and provides a bacteriophage lyase or a chimera and application thereof, and the bacteriophage lyase or the chimera comprises: (1) a natural bacteriophage lyase selected from an amino acid sequence of GNL14, GNL24, GNL25, GNL46, GNL53, GNL66, GNL68, GNL73, GNL80 or GNL82; (2) an antibacterial peptide, wherein the antibacterial peptide is selected from an amino acid sequence of GNP6, GNP24, GNP27, GNP30 or GNP45; and (3) a linker, wherein the linker is selected from the amino acid sequence of GNLK1, GNLK2, GNLK4, GNLK5, GNLK6, GNLK7, GNLK8, GNLK9 or GNLK11. The invention can be used for treating or preventing bacterial infection caused by gram-negative bacteria, especially infection of acinetobacter baumannii, klebsiella and pseudomonas.

Provided are a bacteriophage lytic enzyme or a chimera and a use thereof. The lytic enzyme or chimera thereof comprises: (1) a natural bacteriophage lytic enzyme, the natural bacteriophage lytic enzyme being selected from the amino acid sequences of GNL14, GNL24, GNL25, GNL46, GNL53, GNL66, GNL68, GNL73, GNL80, or GNL82; (2) an antimicrobial peptide, the antimicrobial peptide being selected from the amino acid sequences of GNP6, GNP24, GNP27, GNP30, or GNP45; and (3) a linker, the linker being selected from the amino acid sequences of GNLK1, GNLK2, GNLK4, GNLK5, GNLK6, GNLK7, GNLK8, GNLK9, or GNLK11. The lytic enzyme or chimera thereof can be used for treating or preventing bacterial infections caused by Gram-negative bacteria, particularly infections of Acinetobacter baumannii, Klebsiella, and Pseudomonas.

The present invention discloses a prophage lytic enzyme of Streptococcus equi subspecies equi, and its preparation method and application, which belong to the technical field of veterinary medicine. The amino acid sequence of the lytic enzyme is shown in SEQ ID NO.29. The polynucleotide encoding the prophage lytic enzyme of Streptococcus equi subspecies equi, the expression vector containing the polynucleotide, and the host bacteria containing the expression vector are also within the protection scope of the present invention. Among them, preferably, the sequence of the polynucleotide is shown in SEQ ID NO.30. The present invention compares the prophage lytic enzyme genes in the genome sequence of Streptococcus equi subspecies equi, designs specific primers, amplifies and clones the lytic enzyme genes in the Streptococcus equi strains preserved in the laboratory, performs recombinant expression, and evaluates the antibacterial effects of different lytic enzymes, and finally screens out the lytic enzyme with the best bactericidal effect, thereby providing a new technical means for the prevention and treatment of equine strep.

A bacteriophage lytic enzyme and its application. The name of the bacteriophage lytic enzyme is PlyNJ3; the gene fragment encoding the lytic enzyme PlyNJ3 is amplified from streptococcus suis by PCR, and a prokaryotic expression vector plasmid pET32a-PlyNJ3 containing the PlyNJ3 gene is constructed, the recombinant plasmid is obtained by transforming DH5 alpha competent cells, and the protein is massively expressed in an escherichia coli expression system. Then, the lytic enzyme PlyNJ3 is purified by affinity purification medium, the high-efficiency expression of the lytic enzyme PlyNJ3 in vitro is realized, and the lytic effect of the lytic enzyme PlyNJ3 on different serotypes of streptococcus suis, streptococcus agalactiae and other common streptococci is verified in vitro. The bacteriophage lytic enzyme PlyNJ3 prepared by the application can selectively lyse target pathogenic bacteria, and has a good application prospect in the development of new bacteriophage drugs.

The application provides a microcapsule of prophage lysozyme and a preparation method thereof, and the preparation method comprises the following steps: transforming an expression plasmid containing a recombinant Sply181 lysozyme gene into engineering bacteria, collecting bacterial bodies after culture induction, obtaining lysozyme through ultrasonic crushing, centrifugation and metal ion affinity chromatography purification; preparing the enzyme into a prophage lysozyme mother liquor by mixing with a calcium chloride solution, adding an emulsifier and an anti-acid antioxidant, homogenizing and emulsifying to form a core material suspension; mixing the core material suspension with a sodium alginate-agarose colloidal solution, homogenizing, dropping into a calcium lactate solution through a spraying or extrusion method, solidifying, collecting microcapsule primary bodies and performing low-temperature vacuum drying and shaping; and mixing the dried microcapsule primary bodies with talc, fishy peptide, vanillin and taurine, and then performing a powder coating operation to obtain the microcapsule of prophage lysozyme. The application has the effects of removing endotoxins in engineering bacteria and adding broad-spectrum attractants, and can significantly improve the feeding desire and intake of fish.

The invention provides a prophage lyase microcapsule and a preparation method thereof, and the preparation method comprises the following steps: transforming expression plasmids containing recombinant Sply181 lyase genes into engineering bacteria, collecting thalli after culture induction, and obtaining lyase through ultrasonic disruption, centrifugation and metal ion affinity chromatography purification; preparing the prophage lyase and a calcium chloride solution into prophage lyase mother liquor, adding an emulsifier and an anti-acid and anti-oxidant, and homogenizing and emulsifying to form core material suspension; mixing and homogenizing the core material suspension and the sodium alginate-agarose colloidal solution, dropwise adding the mixture into a calcium lactate solution through a spraying or extrusion method for curing, collecting a microcapsule primary body, and performing low-temperature vacuum drying and shaping; and mixing the dried microcapsule primary body with talcum powder, fishy peptide, vanillin and taurine, and then carrying out powder coating operation to prepare the prophage lyase microcapsule. The feed additive has the effects of removing toxins in engineering bacteria and adding a broad-spectrum phagostimulant, and can remarkably improve the appetite and intake of fishes.

The present application relates to a kind of bacteriophage lytic enzyme LysSPYS2, containing bacteriophage lytic enzyme composition and its application, bacteriophage lytic enzyme LysSPYS2 sequence is as shown in SEQ ID No.1, the bacteriophage lytic enzyme has high-efficiency, broad-spectrum bactericidal effect;It has lytic effect to a variety of bacteria, can be used as bacteriostatic agent;Further, the bacteriophage lytic enzyme is combined with polymyxin B, can exert synergistic antibacterial effect, significantly reduce the amount of antibiotic;The bacteriophage lytic enzyme can also be used with Magnolol, realize outstanding bacteriostatic effect.

This invention relates to a phage lysin LysSPYS1, a composition containing the phage lysin, and their applications. The sequence of the phage lysin LysSPYS1 is shown in SEQ ID No. 1. This phage lysin exhibits highly efficient and broad-spectrum bactericidal activity; it has a lytic effect on a variety of bacteria and can be used as a bacteriostatic agent. Furthermore, combining this phage lysin with polymyxin B can exert a synergistic antibacterial effect, significantly reducing the amount of antibiotics used. The phage lysin can also be used in combination with magnolol to achieve a prominent antibacterial effect.

The invention discloses a preparation method for inducing staphylococcus aureus extracellular vesicles by utilizing phage lyase PlySs2 and application of the staphylococcus aureus extracellular vesicles. The extracellular vesicles are obtained by subjecting staphylococcus aureus in a logarithmic phase or a stable phase to PlySs2 treatment, filtration, ultrafiltration and ultracentrifugation purification. Compared with natural extracellular vesicles, the extracellular vesicles induced by the PlySs2 are higher in yield and good in uniformity, the toxicity is remarkably reduced, and the extracellular vesicles can be used for immunogenicity research and research and development of vaccine compositions.

The invention discloses a protein for enhancing foreign protein expression and application thereof. The target protein-OMP43 recombinant protein can be obtained by fusing and connecting the protein OMP43 to the C end of the target protein, so that the expression level of the target protein in a heterologous expression system is improved. The recombinant protein En6-OMP43 is constructed and obtained by taking bacteriophage lyase En6 as an example. Experimental results show that after the OMP43 is fused, the antibacterial activity of the recombinant protein lyase on acinetobacter baumannii is maintained. The protein OMP43 serving as an expression enhancing element is high in universality, simple and convenient to construct and suitable for being modularly applied to foreign protein expression and preparation.

A phage lyase with an amino acid sequence set forth in SEQ IN NO.2. The lyase can inhibit the growth of Staphylococcus, Escherichia, Klebsiella, and Salmonella bacteria, and can be used as an antibacterial substance in milk pollution control, belonging to the field of bioengineering. The phage lyase can be used for preparing an enzyme formulation alone or in a compounded manner to specifically inactivate bacteria such as Staphylococcus aureus, thereby providing an enzyme formulation source which is safe and free of toxic and side effects for controlling Staphylococcus aureus pollution in milk at present.

The invention relates to the technical field of biology, in particular to listeria monocytogenes bacteriophage lyase Lys039 and application, and the amino acid sequence of the lyase Lys039 is shown as SEQ ID NO.1; the lyase Lys039 has the cracking activity, and shows the cracking activity on all listeria; the molecular marker has a wide splitting spectrum, but cannot split staphylococcus, bacillus subtilis and gram-negative bacteria, and has specificity. The sterilization capability is relatively high, and the concentration dependence is realized; good temperature tolerance is achieved, and the highest temperature can reach 80 DEG C; the listeria monocytogenes strain has the advantages that the listeria monocytogenes strain shows bactericidal activity in the pH range of 3-12, the listeria monocytogenes viable count can be obviously reduced in both acidic and alkaline environments, and when the pH is 6, the listeria monocytogenes strain has higher activity and shows higher acid and alkali resistance.

The utility model provides a device for extracting lysate of obligate bacteriophage of soil ralstonia solanacearum, and relates to the technical field of enrichment and separation of bacteriophage, the device comprises a centrifugal tube and an enrichment tube, the centrifugal tube is provided with a sample inlet and a sampling port, the inside of the centrifugal tube is provided with an accommodating cavity communicated with the sample inlet and the sampling port, and the enrichment tube is arranged in the accommodating cavity. The enriching pipe is detachably mounted in the accommodating cavity, the enriching pipe is provided with a sample receiving port and a filter port, the sample receiving port is communicated with the filter port, the sample receiving port is arranged towards the sample inlet, and a filter screen is arranged at the filter port. According to the utility model, enrichment and separation of bacteriophages can be completed in the same equipment.

The invention discloses a saline-alkaline-tolerant vibrio-inhibiting aquatic probiotic and a preparation process thereof. According to the invention, the gradient domestication culture of the core flora and the directed expression technology of the engineering bacteria realize the active adaptive capacity of the probiotics to the high saline-alkaline environment. After stepped salinity and pH adaptive evolution, the halophilic bacillus and the alkali-resistant lactobacillus can be stably colonized in a culture water body and continuously secrete antibacterial metabolites; and the bacteriophage lyase expressed by the engineering bacteria can accurately destroy the cell wall structure of the vibrio to form a physical-biochemical dual antibacterial barrier. Meanwhile, due to the collaborative design of a functional synergist and ecological auxiliary materials, the flora metabolism activity is optimized through components such as alginate oligosaccharide and a nano-carrier, and a dynamic micro-ecological regulation and control network is constructed by virtue of materials such as modified zeolite and tetrahymena cysts, so that vibrio virulence gene expression is effectively blocked, and environmental toxins are removed; a three-dimensional protection system from strains to an ecological system is formed.