24 results about "Bacillus smegmatis" patented technology

The invention provides recombinant escherichia coli with high yield of ergothioneine and a method for preparing ergothioneine, the recombinant escherichia coli co-expresses ergothioneine anabolic enzymes Egt1 and Egt2, the methyltransferase structural domain of the Egt1 is derived from trichoderma reesei, mycobacterium smegmatis EgtD or rhizopus nigricans, and the methyltransferase structural domain of the Egt2 is derived from the trichoderma reesei, the mycobacterium smegmatis EgtD or the rhizopus nigricans. The histidine sulfoxide synthase structural domain is derived from trichoderma reesei, schizosaccharomyces pombe or rhizopus nigricans, and the Egt2 is derived from claviceps purpurea, neurospora crassa or aspergillus niger. After the recombinant escherichia coli obtained by screening is cultured for 68 hours in a 5L fermentation tank, the yield of ergothioneine can reach 5g / L or above; wherein after recombinant escherichia coli co-expressing Egt1 enzyme (the structural domain of methyltransferase is derived from mycobacterium smegmatis EgtD, and the structural domain of histidine sulfoxide synthase is derived from trichoderma reesei) and Egt2 enzyme (derived from neurospora crassa) are cultured in a 5L fermentation tank for 68 hours, the yield of ergothioneine can reach 8.7 g / L, and the production intensity reaches 127.9 mg / (L.h).

The invention discloses a mycobacteriophage and application thereof, and relates to the technical field of bacteriophages. The mycobacteriophage is named as Mycobacter phage BZNK001, is preserved in Guangdong Microbial Culture Collection Center, has a preservation number of GDMCC No: 67907-B1, does not contain antibiotic drug-resistant genes and integrase genes, has excellent biological safety and good environmental tolerance, can maintain high biological activity in a temperature range of 37-50 DEG C under an acid-base condition of pH 5-9, and can be used for preparing the antibiotic drug-resistant gene and integrase gene-containing mycobacteriophage for preventing and treating the antibiotic drug-resistant gene and integrase gene-containing mycobacteriophage for preventing and treating the antibiotic drug-resistant gene and integrase gene-containing mycobacteriophage. The bacillus subtilis has broad-spectrum and high-efficiency non-tuberculous mycobacterium cracking activity, and can effectively crack four clinical key mycobacteria such as mycobacterium abscessus, mycobacterium avium, mycobacterium intracellularis and mycobacterium smegmatis. According to the invention, a mycobacteriophage resource library is enriched, a novel and stable biological prevention and control means is provided for efficient prevention and control of mycobacteria, and the mycobacteriophage has good clinical application and industrialization values.

The invention relates to a photoactivated carbonyl manganese (I) complex and a preparation method and application thereof in antibiosis. The manganese (I) complex has a good photoactivation antibacterial effect. Specifically, the manganese carbonyl (I) complex can be photoactivated to release carbon monoxide and ligands under the irradiation of white light. By detecting MIC90 of the photoactivated carbonyl manganese (I) complex to different bacteria, the result shows that the photoactivated carbonyl manganese (I) complex has a better antibacterial effect on mycobacterium smegmatis.

Provided herein are Mycobacterium smegmatis porin nanopores, systems that comprise these nanopores, and methods of using and making these nanopores. Such nanopores may be wild-type MspA porins, mutant MspA porins, wild-type MspA paralog porins, wild-type MspA homolog porins, mutant MspA paralog porins, mutant MspA homolog porins, or single-chain Msp porins. Also provided are bacterial strains capable of inducible Msp porin expression.

This invention provides a method for constructing a high-yield ergothionein-producing Bacillus amyloliquefaciens engineered strain and its application. The engineered strain uses the food-grade safe strain Bacillus amyloliquefaciens HZ12 as the starting strain. Through genetic engineering, a key enzyme gene for ergothionein biosynthesis is heterologously expressed in Bacillus amyloliquefaciens. An RBS co-expression strategy is employed to express the mutant gene NcEgt1 derived from Neurospora crassa. Y174F / E51A Co-expression of the EgtE gene derived from *Mycobacterium smegmatis* successfully achieved heterologous synthesis of ergothioneine in *Bacillus amyloliquefaciens*. Under these conditions, the yield of ergothioneine reached 402.68 mg / L, marking the achievement of ergothioneine production in *Bacillus amyloliquefaciens* from scratch. Subsequently, through chassis strain modification, the yield of ergothioneine was further increased to 690.43 mg / L.

Antimicrobial peptides (AMPs), small compounds that often exhibit broad spectrum antimicrobial activity, are garnering interest as potential therapeutics against antibiotic-resistant bacterial pathogens. Development of new AMPs is arduous due to the practical limitations of classical protein-based discovery approaches. A high throughput bioinformatics approach is described which is able to confirm identification of known AMPs from the North American bullfrog (Rana (Lithobates) castebeiana) genome, and a bioinformatics approach is used to develop new AMPs, as described herein as SEQ ID NOs: 1-178. The described AMPs exhibit antimicrobial activity against Mycobacterium smegmatis via microtitre broth dilution assays, indicating broader efficacy.

The application discloses a methyl bacillus subtilis sulfoxide synthase MpEgtB mutant and application thereof, and belongs to the technical field of bioengineering. The application provides a methyl bacillus subutilis-derived sulfoxide synthase MpEgtB mutant capable of efficiently preparing ergothioneine and a modification method of the mutant, and the mutant protein is used to catalyze histidine trimethyl inner salt, L-Cys is used as a sulfur donor to directly convert the histidine trimethyl inner salt into histidine trimethyl inner salt-based cysteine sulfoxide, and the obtained mutant is coupled with Mycobacterium smegmatis-derived MsEgtD and MsEgtE to prepare L-EGT by using L-His and L-Met as substrates. The optimal mutant strain is used as a biological catalyst to perform whole-cell conversion, a continuous feeding strategy is adopted in a 5L fermenter, and the conversion is performed for 48h, so that the yield of L-EGT reaches 8.32g / L. The production process is simple, has few impurities, and has an important industrial application prospect.

The present invention relates to a photoactivated manganese (I) carbonyl complex, its preparation method, and its use in antibacterial applications. The manganese (I) complex exhibits significant photoactivated antibacterial effects. Specifically, under white light irradiation, the manganese (I) carbonyl complex can be photoactivated to release carbon monoxide and a ligand. Testing of the MIC90 values ​​of the photoactivated manganese (I) carbonyl complex against various bacteria revealed that the complex exhibited significant antibacterial effects against Mycobacterium smegmatis.

Provided herein are mutant single-chain Mycobacterium smegmatis porin (Msp) and uses thereof.

The invention discloses mycobacterium bacteriophage bactericidal protein and a preparation method thereof. The preparation method comprises the following steps: S1, constructing a bacteriophage gene gp48 induced expression strain; s2, determining a phage gene expression strain; s3, constructing a phage metallophosphatase induced expression strain; s4, purifying the phage metal phosphatase protein; according to the invention, a metal phosphoesterase is identified in a bacteriophage genome with relatively strong lysis activity on mycobacterium tuberculosis and mycobacterium smegmatis, and the expression of the metal phosphoesterase has relatively strong killing activity on bacteria and relatively strong phosphodiesterase activity in vitro; the invention also provides a safe and convenient protein preparation method of the phage metal phosphatase.

The invention discloses application of a caragana crude extract and an n-butanol layer extract in antituberculous activity and preparation methods of the caragana crude extract and the n-butanol layer extract, and the preparation method of the caragana crude extract comprises the following steps: weighing dried and crushed caragana roots, adding 90% absolute ethyl alcohol with the same volume as the caragana roots, extracting by adopting a heating reflux extraction method, and concentrating under reduced pressure to obtain the caragana crude extract. Sequentially extracting with petroleum ether, dichloromethane and n-butyl alcohol to obtain caragana petroleum ether layer extract, dichloromethane layer extract and n-butyl alcohol layer extract; tests prove that the caragana crude extract and the caragana n-butyl alcohol layer extract have remarkable bacteriostatic activity on mycobacterium smegmatis, have a strong bacteriostatic effect on a mycobacterium tuberculosis standard strain H37Rv (ATCC27294) and have certain bacteriostatic activity on a multidrug-resistant tubercle bacillus strain Mtb706, and by means of activity-oriented extraction process optimization, the content of the caragana n-butyl alcohol layer extract is greatly improved. The screening range of the antituberculous active ingredients in the caragana roots is effectively narrowed, and a key foundation is laid for the subsequent separation and purification of the active ingredients and the research on the action mechanism.

The invention discloses application of an STX6 gene as a target spot in prevention and treatment of tuberculosis, the survival of mycobacterium bovis, mycobacterium tuberculosis, BCG and mycobacterium smegmatis in host cells can be remarkably inhibited by knocking out or knocking down the expression of the STX6 gene, the host cell death induced by the mycobacterium bovis and the mycobacterium tuberculosis is inhibited, and the normal proliferation of the host cells is not influenced. Meanwhile, the STX6 is highly conserved in mammals, so that the STX6 can be widely applied to prevention and treatment of tuberculosis caused by humans and animals, can also be used as a target for gene editing animal design, is applied to genetic therapy and disease-resistant breeding, and has rich application and economic values.

A method for endogenously extracting a Mycobacterium smegmatis protein nanocage is provided. The method includes the following steps: introducing a recombinant plasmid containing a CFP29 gene and a 1×Flag affinity tag into a Mycobacterium smegmatis strain to obtain a recombinant Mycobacterium smegmatis strain; extracting a total protein solution of the recombinant Mycobacterium smegmatis strain, and subjecting the total protein solution of the recombinant Mycobacterium smegmatis strain to Flag tag affinity column chromatography purification to obtain a crude extract of the Mycobacterium smegmatis protein nanocage; and subjecting the crude extract of the Mycobacterium smegmatis protein nanocage to gel exclusion chromatography purification to obtain a pure product of the Mycobacterium smegmatis protein nanocage. The method is simple to operate and convenient to implement during extraction and purification, and an obtained Mycobacterium smegmatis background protein nanocage has a high yield, excellent purity, and stable properties.

Provided herein are mutant single-chain Mycobacterium smegmatis porin (Msp) and uses thereof.

The invention discloses a glutamate decarboxylase mutant and application thereof, and belongs to the technical field of biology. Glutamic acid decarboxylase BGAD from escherichia coli and a glutamic acid decarboxylase mutant MsGAD obtained by mutating glutamic acid decarboxylase from mycobacterium smegmatis are used for catalyzing L-glutamic acid to synthesize gamma-aminobutyric acid, and high-purity and high-yield gamma-aminobutyric acid can be obtained; in the catalytic reaction, hydrochloric acid or sulfuric acid does not need to be additionally added to regulate pH, almost no L-glutamic acid is left after catalysis is finished, the yield reaches 98% or above, and the yield of gamma-aminobutyric acid can reach 605g / L or above; meanwhile, the production process provided by the invention is simple and easy to operate, can be applied to large-scale industrial production, and has a good application prospect.

The invention discloses a mycobacterium protein with an anti-inflammatory effect as well as preparation and application of the mycobacterium protein. The invention discloses a mycobacterium protein Rv2723 with an amino acid sequence as shown in SEQ ID NO.1. The protein of the sequence is reported for the first time. Activity studies show that the Rv2723 recombinant protein can significantly inhibit the expression of LPS-induced macrophage inflammatory factors, the Rv2723 overexpression can effectively inhibit the expression of Mycobacterium smegmatis-induced macrophage inflammatory factors, and the Rv2723 overexpression can down-regulate the expression of Mycobacterium smegmatis-infected mouse serum inflammatory factors IL-6. In addition, the Rv2723 does not generate excessive immunological rejection. Therefore, the mycobacterium protein Rv2723 with the amino acid sequence as shown in SEQ ID NO.1, the coding gene of the mycobacterium protein Rv2723, and a recombinant vector, recombinant bacteria or recombinant cells containing the coding gene have the prospect of being developed into anti-inflammatory drugs.

The invention relates to an exosome-coated phosphorus-containing dendrimer / antibacterial peptide composite material as well as preparation and application thereof. The exosome-coated phosphorus-containing dendrimer / antibacterial peptide composite material comprises a phosphorus-containing dendrimer, a peptide and an exosome. The preparation method disclosed by the invention has the characteristics of simplicity, high reaction process controllability, easiness in operation and separation and the like, the intracellular transmission efficiency and antibacterial activity of nisin can be improved, efficient removal of mycobacterium smegmatis is realized by virtue of an exosome-mediated host targeting function, and the preparation method has a good development prospect and application value in treatment of pulmonary tuberculosis patients.

The utility model relates to the technical field of bacteria shaking tubes, and discloses a mycobacterium smegmatis bacteria shaking tube. The mycobacterium smegmatis shaking tube comprises a test tube main body, the stirring plate is positioned in the test tube main body and is used for improving the shaking efficiency of mycobacterium smegmatis; a cover is arranged at the top of the test tube body and connected with the top of the test tube body in a clamping mode, a plug is arranged in the cover, the top of the plug and the cover are integrally formed, and a rotating rod rotates on a supporting block through a connecting block. The stirring plate rotates to stir the mycobacterium smegmatis precipitated at the bottom of the test tube main body, so that the condition that the mycobacterium smegmatis is precipitated when the mycobacterium smegmatis is centrifugally shaken through the test tube main body is prevented, and meanwhile, the mycobacterium smegmatis is stirred through the stirring plate, so that the shaking efficiency of the mycobacterium smegmatis is improved; therefore, the working efficiency is improved, and the practicability of the device is embodied.

The present invention belongs to the field of organic synthesis technology and specifically provides a photoactive ruthenium complex and a preparation method thereof. The photoactive ruthenium complex provided by the present invention not only decomposes and releases isoniazid under light but also generates singlet oxygen under light excitation, exhibiting dual antibacterial effects, including significant antibacterial effects against Mycobacterium smegmatis, and can be used to treat tuberculosis infection.

This invention belongs to the field of microbiology, specifically relating to the application of albizophenolic compounds. These compounds inhibit pathogenic bacteria such as Salmonella, Mycobacterium smegmatis, Enterococcus faecalis, and Staphylococcus aureus, and inhibit the growth of human lung cancer cells and human breast cancer cells, exhibiting bactericidal and anticancer activities. They are suitable for developing products for everyday household use, medical care, and animal husbandry, with broad prospects for industrial application.

The invention discloses application of a PAPSS2 gene as a target spot in prevention and treatment of tuberculosis, the PAPSS2 gene is knocked out or knocked down, so that survival of mycobacterium bovis, mycobacterium tuberculosis, BCG and mycobacterium smegmatis in host cells can be remarkably inhibited, and normal proliferation of the host cells is not affected. Meanwhile, the PAPSS2 is highly conserved in mammals, so that the PAPSS2 can be widely applied to prevention and treatment of tuberculosis caused by humans and animals, can also be used as a target for gene editing animal design, is applied to genetic therapy and disease-resistant breeding, and has rich application and economic values.

The invention belongs to the technical field of biological medicine, provides a method for constructing a biosafety mouse bone tuberculosis model based on mycobacterium smegmatis, and evaluates that under the condition of a standard biosafety level (BSL)-1, the mycobacterium smegmatis induces local and disseminated bone infection through three infection pathways, and the bone tuberculosis model has the advantages that the bone tuberculosis is induced by the mycobacterium smegmatis through three infection pathways; results show that the mouse model for intracentral injection administration of the three infection pathways is closer to the latent progress of human bone tuberculosis. The mouse model based on the mycobacterium smegmatis provides a simple and reliable platform related to immunopathology for exploring host-pathogen dynamics, immunologically-driven bone destruction and early treatment tests.

The application relates to the field of traditional Chinese medicine against tuberculosis, in particular to a compound extracted from Magnolia officinalis, a preparation method and application of the compound, the compound is shown as formula I, the compound can effectively inhibit the growth of Mycobacterium smegmatis, and can prevent and treat Mycobacterium smegmatis infection, and the MIC of the compound on Mycobacterium smegmatis is 60 ug / mL.