10 results about "Lysostaphin" patented technology

Unglycosylated lysostaphin variant protein, nucleic acid molecule, vector and host cell, as well as a method for production of unglycosylated lysostaphin variant protein in a yeast expression system are provided. The proteins are produced in a Pichia pastoris expression system and have been shown to have activity equivalent to wild-type lysostaphin. The lysostaphin variant proteins can be used as therapeutic proteins for treatment of diseases such as Staphylococcus aureus infection.

When bacteria with different bacteriolysis targets are detected, it was necessary to lyse the individual bacteria separately with lysis solutions containing lysis enzymes suitable for each bacteriolysis target, resulting in a complicated and delayed bacteriolysis process. The purpose of this invention is to provide a new bacteriolysis method for lysing bacteria belonging to two or more species in a sample, making the bacteriolysis process more efficient and rapid.Provided is method for lysing a group of bacteria in the sample, comprising the step of lysing the group of bacteria in the sample in a mixture solution obtained by mixing the sample, a bacteriolysis enzyme, and a bacteriolysis aid, wherein the bacteriolysis enzyme includes at least one bacteriolysis enzyme selected from the group consisting of lysostaphin, lysozyme, acetyl glucosaminidase, and endopeptidase, and wherein the mixture solution has a pH of from 6.0 to 7.0 and an electrical conductivity of from 2.5 to 8.5 mS / cm, whereby bacteriolysis of two or more species of bacteria becomes possible.

The invention discloses a method for separating, screening and identifying staphylococcus in a chick embryo sample, which comprises the following steps: S1, sterilizing collected chick embryo tissues, cutting into pieces, mixing with an SCB liquid culture medium, and culturing in a shaking table to obtain a culture solution; s2, scribing the obtained culture solution into a BP (Back Propagation) culture plate, putting the plate into an incubator for standing culture, after monoclone grows out of a culture dish, picking the monoclone, and uniformly blowing and beating the monoclone in PBS (Phosphate Buffer Solution) to obtain a mixed bacteria solution; s3, incubating the obtained mixed bacteria solution in lysostaphin to obtain a mixed solution I; s4, Tris-HCl and protease K are added into the obtained mixed solution I, incubation is conducted firstly, then heating is conducted, and a mixed solution II is obtained; and S5, carrying out PCR amplification by taking the obtained mixed solution II as a template to obtain a product, and identifying. The method is accurate, rapid, economical and efficient.

The invention discloses a nano-motor composite material with an eccentric core-spinous shell structure and a preparation method and application thereof.The nano-motor composite material is of a core-shell structure, the core of the nano-motor composite material is composed of a carbon material, the space between the core and the shell is filled with indocyanine green and lysostaphin materials, the shell is composed of a SiO2 carbon material, and the nano-motor composite material is of a spinous porous structure; indocyanine green and lysostaphin are loaded on the surface; and the sphere centers of the core and the shell do not coincide. The nano-motor composite material disclosed by the invention is combined with the photosensitizer ICG and the carbon material in the material under the irradiation of NIR, and the asymmetric eccentric structure of the nano-motor composite material can enable the composite material to be non-uniformly heated to generate self-heating driving force, so that the targeting effect of lysostaphin and ROS on an MRSA biological membrane is realized. The nano motor is applied to preparation of drugs for promoting healing of diabetic wounds, has excellent tissue infiltration capacity and drug delivery efficiency, and shows efficient synergistic antibacterial performance.

The invention provides lysostaphin with high thermal stability as well as a preparation method and application thereof, and belongs to the field of gene engineering and enzyme engineering. Specifically, a plurality of amino acid residue pairs possibly forming disulfide bonds are predicted on the basis of structural analysis of lysostaphin and assistance of various thermal stability prediction software. And verifying and screening to obtain a plurality of mutants with improved thermal stability. Therefore, the method has good practical use value.

The invention relates to the technical field of biology, and particularly discloses a phage-sourced staphylococcus aureus antibacterial peptidase lysolytic protein mutant with high thermal stability as well as a preparation method and application of the phage-sourced staphylococcus aureus antibacterial peptidase lysolytic protein mutant. According to the mutant, stability design is carried out on wild type staphylococcus aureus antibacterial peptidase lysostaphin (APL) through a PROSS online prediction tool, nine mutation schemes (design 1-design 9) are obtained, the thermal stability of the mutant APL (design 9) is remarkably improved, the optimal temperature is increased to 55 DEG C, 70 DEG C and 80 DEG C, and the half-life period reaches 57 + / -4 min and 21 + / -1 min respectively and is 1.4 times and 2.1 times of the half-life period of the wild type. The preparation method comprises the steps of gene synthesis, vector construction, recombinant expression and purification. The mutant can be used for preventing and treating staphylococcus aureus infection, veterinary antibiotic substitutes, food preservatives and compound disinfectants, is suitable for a high-temperature processing environment, and has industrialization advantages.

The application relates to the technical field of biotechnology, and particularly discloses a bacteriophage-derived lysostaphin mutant with high thermal stability and an application and a preparation method thereof. The mutant is obtained by using a PROSS online prediction tool to design the stability of wild-type antimicrobial peptidase lysostaphin (APL), and nine mutation schemes (design 1-design 9) are obtained. The thermal stability of the mutant APL (design 9) is significantly improved, the optimum temperature is increased to 55 DEG C, and the half-life at 70 DEG C and 80 DEG C is 57+ / -4 min and 21+ / -1 min, respectively, which is 1.4 times and 2.1 times that of the wild type. The preparation method comprises gene synthesis, vector construction, recombinant expression and purification. The mutant can be used for the prevention and treatment of staphylococcus aureus infection, veterinary antibiotic substitutes, food preservatives and compound disinfectants, and is suitable for high-temperature processing environment and has industrialization advantages.

The invention provides a method for improving the expression quantity of bacterial strain lysostaphin, belongs to the technical field of biology, and particularly relates to a method for improving the expression quantity of the bacterial strain lysostaphin by transferring a plasmid pET-28a-lacUV5 (49)-T7RNAP and a plasmid pET-22b-lys containing a coding lysostaphin gene lys into an expression bacterial strain EcN P so as to improve the expression quantity of the bacterial strain lysostaphin. The nucleotide sequence of the plasmid pET-28a-lacUV5 (49)-T7RNAP is shown as SEQ ID NO.1, the amino acid sequence of the lysostaphin is shown as SEQ ID NO.2, and the expression strain EcN P is Escherichia coli Nissle1917 of which the plasmids pMUT1 and pMUT2 are knocked out.

The application provides a recombinant Escherichia coli Nissle 1917 for producing lysostaphin, and belongs to the technical field of biotechnology.The recombinant Escherichia coli Nissle 1917 comprises an expression strain EcN△P, a plasmid pSC101- lacUV5-T7RNAP and a plasmid pET22b- lys containing a lysostaphin gene lys coding sequence. lacUV5-T7RNAP The nucleotide sequence of the plasmid pSC101- Escherichia coli is SEQ ID NO.1; the amino acid sequence of the lysostaphin is SEQ ID NO.2; the expression strain EcN△P is a knock-out plasmid pMUT1 and pMUT2 Escherichia coli Nissle 1917; the application provides a recombinant Escherichia coli Nissle 1917 for producing lysostaphin, and the recombinant bacteria produce lysostaphin with high protein concentration.

This invention discloses an engineered Trichoderma reesei strain that secretes and expresses lysostaphin, its construction method, and its applications, belonging to the field of genetic engineering technology. This invention obtains the lysostaphin proenzyme and its encoding gene by modifying lysostaphin with a secretory peptide. The encoding gene, promoter, and terminator are then ligated into a shuttle plasmid to obtain a lysostaphin expression plasmid. This plasmid is transformed into Trichoderma reesei, and the endogenous protein encoding gene cbh1, which has the highest secretion expression level in Trichoderma reesei, is replaced with the aforementioned encoding gene, thus constructing an engineered Trichoderma reesei strain that secretes and expresses lysostaphin. This invention is the first successful construction of an engineered Trichoderma reesei strain capable of secreting and expressing lysostaphin, which simplifies the production process of lysostaphin, significantly reduces its production cost, and lays the foundation for large-scale industrial production of lysostaphin.