66 results about "Clostridium organisms" patented technology

The invention discloses a camel source single-domain antibody GH12 for recognizing a conformation B cell epitope in clostridium difficile glutamate dehydrogenase and application of the camel source single-domain antibody GH12. The nucleotide sequence of the camel source single-domain antibody GH12 is shown as SEQ ID NO: 1, and the amino acid sequence of the camel source single-domain antibody GH12 is shown as SEQ ID NO: 2. The camel source single-domain antibody GH12 disclosed by the invention has the advantages of strong specificity, high sensitivity, good accuracy and the like, and it is proved that the camel source single-domain antibody GH12 can be used for detecting recombinant expressed CD-GDH and endogenous CD-GDH by enzyme-linked immunosorbent assay (ELISA), protein dot blot hybridization (DB) and other methods.

The invention relates to the technical field of microorganisms, in particular to application of paraclostridium benzene to promotion of degradation of methyl mercury. The preservation number of the paraclostridium behenolyticum is KCTC (Korean Center of Type Culture Collection) 15476 or LMG (Leukocyte Molecular Growth) 28745, and the preservation number of the paraclostridium behenolyticum is KCTC 15476. The invention discloses the capability of degrading the toxic pollutant methyl mercury of the paraclostridium behenoelyticum for the first time, and the paraclostridium behenoelyticum has important significance for deeply knowing the form transformation effect of microorganisms on mercury, understanding the biogeochemical cycle of mercury and repairing and treating microorganisms polluted by methyl mercury in the environment.

Provided are methods and compositions for treating and / or preventing C. difficile infections, particularly recurring C. difficile infections. The compositions for use in treating and / or preventing C. difficile infections include in some embodiments at least one agent that inhibits an alpha 2 adrenergic receptor, and the presently disclosed methods include administering at least one such composition to a subject in need thereof, optionally in combination with other therapeutically active agents including but not limited to an enhancer of an IL-13 biological activity, optionally an IL-13 peptide or a fragment or homolog thereof; an interleukin-13 receptor subunit alpha-2 (IL-13Ra2) inhibitor; an enhancer of an Interleukin-33 (IL-33) biological activity, optionally an IL-33 polypeptide or a biologically active fragment or homolog thereof; or any combination thereof.

The application relates to the field of biotechnology, in particular to a Faecalibacterium prausnitzii microbial anti-inflammatory molecule and a screening method and application thereof. The amino acid sequence of the Faecalibacterium prausnitzii microbial anti-inflammatory molecule is shown in (a) or (b); (a) the amino acid sequence is shown in SEQ ID NO. 1; (b) a protein derived from (a) with anti-inflammatory activity obtained by substituting, deleting or adding one or more amino acids in the amino acid sequence in (a). Compared with the microbial anti-inflammatory molecule protein of a representative strain A2-165 of the Faecalibacterium genus, the inhibiting effects of the Faecalibacterium prausnitzii microbial anti-inflammatory molecule on the NF-kappa B signal pathway and the inflammatory factor TNF alpha are 50% and 56% respectively, and the anti-inflammatory effect (inhibiting the NF-kappa B signal pathway) of the Faecalibacterium prausnitzii microbial anti-inflammatory molecule screened in the application can reach more than 90%, and the Faecalibacterium prausnitzii microbial anti-inflammatory molecule has the advantages of small use dosage and strong anti-inflammatory effect.

The invention relates to the technical field of pet feed additives, and discloses a pet coated enzyme-containing metagen feed additive and a preparation method thereof, the additive comprises a core particle formed by mixing a metagen and a slow-release coated compound enzyme, and an enteric layer coated outside the core particle. The preparation method comprises the following steps: respectively preparing the metagen and the slow-release coated compound enzyme, mixing the metagen and the slow-release coated compound enzyme to prepare core particles, and then carrying out enteric coating on the core particles. The activity of functional components is protected through a double coating structure, so that the functional components are released in an intestinal tract in a targeted manner and have a synergistic effect. The additive can effectively improve pet protein digestion, regulate intestinal microenvironment, remarkably proliferate beneficial bacteria such as lactobacillus and clostridium praeparatum and inhibit escherichia coli, so that soft stool is reduced, and intestinal health of pets is maintained.

The invention discloses a mutant of glutamate dehydrogenase derived from clostridium symbiosum, and an application of the mutant in synthesis of L-glufosinate-ammonium, and particularly discloses a mutant of glutamate dehydrogenase derived from clostridium symbiosum and an application of the mutant in synthesis of L-glufosinate-ammonium. According to the invention, single-site or multi-site mutation is carried out on wild-type glutamate dehydrogenase derived from clostridium symbiotic as shown in SEQ ID NO.1 by virtue of a mutation transformation method which is designed by carrying out rational analysis on amino acid residues in a binding pocket of glutamate dehydrogenase and a substrate, a geometrical shape in a molecular tunnel and interaction of domains in a structural domain; according to the obtained single-site or multi-site mutant, the catalytic activity to 2-carbonyl-4-(hydroxymethylphosphonyl) butyric acid is remarkably improved, and the mutant has an application prospect in preparation of L-glufosinate-ammonium through biological catalysis.

The invention belongs to the technical field of biology, and particularly relates to a Sardinia clostridium 7alpha-hydroxysteroid dehydrogenase mutant S161G. The amino acid sequence of the mutant S161G is as shown in SEQ ID NO: 2, and the amino acid sequence of the mutant S161G is obtained by mutating Ser at the 161st site of a wild type sequence SEQ ID NO: 1 into Gly. The catalytic efficiency of the mutant S161G on substrates NADP < + > and TCDCA is 3.82 times that of a wild type. The mutant S161G provided by the invention has huge application potential in the industrial production process of obtaining TUDCA or UDCA through biotransformation.

Form-specific cell based assays for the characterization of Clostridia botulinum neurotoxins and genetically modified cells utilized in such assays are described. Such assays and genetically modified cells can discriminate between complexed and non-complexed forms of botulinum neurotoxin having the same serotype, for example serotype E. Methods are also provided for generating and identifying cell lines that can be utilized in such cell based assays.

A piezoelectric plate sensor comprising a piezoelectric layer; two electrodes; and an insulation layer. The insulation layer is produced by soaking the piezoelectric layer and two electrodes in a mercaptopropyltrimethoxysilane solution with an amount of water from 0.1 v / v. % to about 1 v / v % and at pH from about 8 to about 150 for a period from about 8 to about 15 hours, and the mercaptopropyltrimethoxysilane solution has a concentration of mercaptopropyltrimethoxysilane from about 0.01 v / v % to about 0.5 v / v %. A method of detecting a biomolecule in a sample using the piezoelectric plate sensor in particular, that of detecting a genetic marker with PCR sensitivity and specificity without the need of DNA isolation or amplification is also provided. The piezoelectric plate sensor may be used to diagnose various diseases including breast cancer, myocardial infarction, diarrhea, Clostridium difficile infection, and hepatitis B infection.

The invention discloses a mutant of glutamate dehydrogenase derived from clostridium symbiosum, and an application of the mutant in synthesis of L-glufosinate-ammonium, and particularly discloses a mutant of glutamate dehydrogenase derived from clostridium symbiosum and an application of the mutant in synthesis of L-glufosinate-ammonium. According to the invention, single-site or multi-site mutation is carried out on wild-type glutamate dehydrogenase derived from clostridium symbiotic as shown in SEQ ID NO.1 by virtue of a mutation transformation method which is designed by carrying out rational analysis on amino acid residues in a binding pocket of glutamate dehydrogenase and a substrate, a geometrical shape in a molecular tunnel and interaction of domains in a structural domain; according to the obtained single-site or multi-site mutant, the catalytic activity to 2-carbonyl-4-(hydroxymethylphosphonyl) butyric acid is remarkably improved, and the mutant has an application prospect in preparation of L-glufosinate-ammonium through biological catalysis.

This invention belongs to the field of biotechnology and discloses a monoclonal antibody that specifically binds to the recombinant protein of Clostridium perfringens (Cp) β2 toxin, a hybridoma cell line, and its applications. This invention provides a monoclonal antibody that specifically binds to the β2 toxin protein and a hybridoma cell line CPB2-2A4 (accession number CCTCC NO: C2024312) that secretes this monoclonal antibody. The monoclonal antibody reacts only with the Cp β2 toxin protein and does not cross-react with other pathogens infecting cattle and sheep, exhibiting extremely high specificity. The kit utilizes competitive ELISA technology, using a mouse monoclonal antibody that specifically binds to the Cp β2 toxin recombinant protein as a competing antibody, enabling the detection of Cp β2 toxin antibodies in serum from various animal sources, avoiding species limitations of sample sources. Furthermore, the kit exhibits good reproducibility. The kit has relatively relaxed technical requirements during operation and can be widely used in production for applications such as Cp epidemiological surveys and vaccine immunization level monitoring.

This present invention provides C-TAB.G5 and C-TAB.G5.1 isolated polypeptides comprising the receptor binding domains of C. difficile toxin A and toxin B as set forth in the amino acid sequences of SEQ ID NO: 2 and SEQ ID NO: 4. The C-TAB.G5 and C-TAB.G5.1 isolated polypeptides may be used to neutralize toxic effects of C. difficile toxin A and / or toxin B.

The present invention relates to the technical field of vaccine preparation, and in particular to an ASP-ETX fusion protein and a Clostridium perfringens epsilon toxin nanoparticle vaccine and a preparation method thereof. The ASP-ETX fusion protein of the present invention comprises Clostridium perfringens epsilon toxin and an activation-related secretory protein; the amino acid sequence of the ASP-ETX fusion protein is shown in SEQ ID NO.1. The Clostridium perfringens epsilon toxin nanoparticle vaccine is obtained by co-incubating the ASP-ETX fusion protein with cell membrane-wrapped nanoparticles. The vaccine exhibits excellent biosafety in both cells and animals, and induces the production of up to 10 5 ETX-specific antibodies with a neutralizing antibody titer of up to 64 can protect mice against 100×LD 50 GST-ETX toxin attack has great potential to replace traditional ETX vaccines.

C. difficile is a leading cause of antibiotic-associated diarrhea and nosocomial infection in the United States. The symptoms of C. difficile infection (CDI) are associated with the production of two homologous protein toxins, TcdA and TcdB, and the toxins are considered bone fide targets for clinical diagnosis as well as the development of novel prevention and therapeutic strategies. Disclosed herein are nanobodies that bind specific structural and functional domains of TcdA and TcdB.

The invention provides antibacterial protein targeting clostridium scintillator as well as a preparation method and application of the antibacterial protein. Specifically, the invention provides an antibacterial protein targeting clostridium scintillator or a chimeric compound thereof, and the antibacterial protein comprises one or more of proteins formed by the following amino acid sequences: (a) an amino acid sequence as shown in any one of SEQ ID NO: 1 to SEQ ID NO: 13; (b) a derivative sequence which is obtained by replacing, adding and / or deleting one or more amino acids from the amino acid sequence of (a) and has the same function as (a); and (c) a derived amino acid sequence of which the identity with any one sequence from SEQ ID NO: 1 to SEQ ID NO: 13 is 85% or more or 90% or more. The antibacterial protein disclosed by the invention can be used for efficiently killing clostridium scintillation and has specificity.

The invention discloses a nucleic acid composition for rapidly detecting type B clostridium botulinum and application of the nucleic acid composition, the composition is composed of a primer pair and a probe formed by single-chain molecules with the name of BoNT / B-P. The primer pair is composed of single-chain DNA with the name of BoNT / B-F3 and single-chain DNA with the name of BoNT / B-R3. According to the invention, a real-time fluorescence RPA rapid detection system of the B-type clostridium botulinum is established, and a new technical support is provided for on-site rapid diagnosis and epidemic disease investigation of the B-type clostridium botulinum.

The application provides a bacteriostatic and deodorizing composite preparation for pets and application thereof, which comprises 30-40% of enzyme preparation, 20-30% of plant extract, 20-30% of probiotic powder, and 0-10% of inorganic salt and filling aid. The application can efficiently remove ammonia and effectively reduce the ammonia gas yield in excrement; can efficiently inhibit E. coli, Salmonella, MRSA, C. perfringens and the like; can reduce fecal odor and effectively reduce fecal odor substances in pet excrement, such as indole, cadaverine, spermidine, putrescine and skatole and the like; is safe and harmless, avoids the negative influence of chemical synthetic substances on pet health, prevents clumping, and maximally reduces the influence on cat litter clumping, thereby guaranteeing the use effect.

The invention provides a subunit vaccine and application thereof in resisting avian clostridium perfringens, and the subunit vaccine is characterized in that non-toxic CPA and NetB proteins are truncated, and three gene sequences of auxiliary protein metallopeptidase are connected in series and fused through Linker to obtain a recombinant gene. The recombinant protein with good solubility and high expression quantity can be obtained from the recombinant gene under the induction conditions that the IPTG concentration is 0.2 mM and the temperature is 25 DEG C. Immunogenicity detection on the N-C-M recombinant protein finds that the N-C-M recombinant protein can induce SPF chicken to generate antibodies. Immune protection test results show that a subunit vaccine prepared from the N-C-M protein and an adjuvant finds that 100% immune protection effect can be achieved for immunizing 100 [mu] g / chicken and 200 [mu] g / chicken, and the subunit vaccine can be used as a subunit vaccine for resisting various toxin types.

The present invention discloses a camel-derived single-domain antibody GA4 that recognizes a linear B-cell antigen epitope in Clostridium difficile glutamate dehydrogenase and its application. The nucleotide sequence of the camel-derived single-domain antibody GA4 is shown in SEQ ID NO: 1, and the amino acid sequence is shown in SEQ ID NO: 2. The camel-derived single-domain antibody GA4 of the present invention has the advantages of strong specificity, high sensitivity, and good accuracy. It has been demonstrated that it can be used for detection of recombinantly expressed CD-GDH and endogenous CD-GDH by methods such as enzyme-linked immunosorbent assay (ELISA) and western blot hybridization (WB).

A method for determining cleavage of a VAMP by a clostridial neurotoxin in a cell that has been contacted with the clostridial neurotoxin under conditions suitable for clostridial neurotoxin activity, the method comprising contacting the cytoplasmic content of the cell with an antibody that binds to a resulting C-terminal product of such cleavage under suitable conditions in vitro or ex vivo and detecting the binding of the antibody to the C-terminal cleavage product. The antibody may, for example, be capable of binding an antigenic polypeptide consisting of 10 to 65 amino acid residues and comprising an epitope comprising an amino acid sequence that is at least 90% identical to an amino acid sequence of at least 8 amino acid residues that is immediately C-terminal to a clostridial neurotoxin cleavage site in the VAMP.

The invention belongs to the technical field of biological enzyme engineering, and particularly relates to a D-psicose 3-epimerase mutant derived from rumen clostridium and application of the D-psicose 3-epimerase mutant. The mutant disclosed by the invention is obtained through specific single-point mutation on the basis of a wild type amino acid sequence of D-psicose 3-epimerase, and specifically, the mutant is obtained through mutation of the following five sites: L159V, Q164E, N238T, E288G and C289I; wherein mutants containing all five single point mutations are named as XT mutants. The mutants significantly improve the thermal stability of the enzyme and the catalytic conversion rate under high substrate concentration, especially under the high substrate concentration of 300-700 g / L, the conversion rate of D-psicose of the XT mutant is improved by 2-3% compared with that of a wild type, and the thermal stability of the XT mutant is improved by 28 times compared with that of the wild type. The mutant is suitable for a high-temperature and high-substrate-concentration industrial conversion system, the production cost of D-psicose can be remarkably reduced, and the mutant has important industrial application value.

The invention discloses a camel source single-domain antibody CB3 for recognizing linear B cell epitope in clostridium difficile methylase CamA and application of the camel source single-domain antibody CB3. The nucleotide sequence of the camel source single-domain antibody CB3 is as shown in SEQ ID NO: 1, and the amino acid sequence of the camel source single-domain antibody CB3 is as shown in SEQ ID NO: 2. The camel source single-domain antibody CB3 disclosed by the invention has the advantages of strong specificity, high sensitivity, good accuracy and the like, and it is proved that the camel source single-domain antibody CB3 can be used for detecting recombinant expressed CD-CamA and endogenous CD-CamA by methods such as enzyme-linked immunosorbent assay (ELISA) and western blot hybridization (WB).

The invention relates to the technical field of gene engineering and enzyme engineering, in particular to a galactosidase mutant and application thereof in production of human milk oligosaccharide. According to the galactosidase mutant, threonine at the 192nd position of galactosidase which is derived from pseudoclostridium thermosuccinate and has an amino acid sequence shown as SEQ ID NO: 1 is changed into tryptophan, or aspartic acid at the 193rd position of the galactosidase is changed into proline, or methionine at the 194th position of the galactosidase is changed into alanine, or methionine at the 195th position of the galactosidase is changed into threonine, or aspartic acid at the 193rd position of the galactosidase is changed into proline. Or the aspartic acid at the 237 position is changed into asparagine. The galactosidase mutant obtained by the invention not only has efficient enzymatic activity, but also can maintain the enzymatic activity at a relatively high temperature, and is suitable for catalyzing the synthesis of human milk oligosaccharide.

The present disclosure is directed to fusion proteins and propeptide fusions comprising a diphtheria toxin catalytic domain and translocation domain with a receptor binding domain from a Clostridial neurotoxin. The fusion protein has a therapeutic cargo positioned upstream of the catalytic domain. Also disclosed are therapeutic agents, treatment methods, propeptide fusions, isolated nucleic acid molecules, expression systems, host cells, and methods of expressing fusion proteins.

The invention relates to the technical field of biosensing and medical instruments, in particular to an organic electrochemical transistor sensor and a preparation method and application thereof. According to the specific technical scheme, the surface of a grid electrode is sequentially modified with a mercaptoacetic acid self-assembly layer, an anti-clostridium difficile toxin B (TcdB) antibody (such as E3), ethanolamine and bovine serum albumin (BSA), and a biological function interface with high specific recognition and strong non-specific adsorption resistance is constructed. The preparation method is simple and stable. The obtained sensor has extremely high detection sensitivity (fg / mL level), good selectivity and a wide linear range on TcdB, can complete detection within a few minutes, is particularly suitable for rapid and accurate detection of TcdB in a complex biological sample, and has a wide application prospect in the field of bedside diagnosis of clostridium difficile infection.

The application discloses a kind of non-small cell lung cancer prognosis immunity and nutrition state related intestinal microorganism marker and its application, intestinal microorganism marker specifically includes: Eubacterium hallii, Akkermansia, Lachnospira pectinosus, Lachnospira viviparous, Paraclostridium, Haemophilus influenzae.The relative abundance of intestinal flora is used with patient PNI index, and the correlation of the diversity of intestinal flora, intestinal type and specific strain and PNI is established.The relative abundance of 6 specific strains of intestinal flora is used with patient PNI index to construct the evaluation model of nutrition immunity, by using this model, the relative abundance information of 6 specific strains of new individual is input, and the result of nutrition immunity evaluation is output, without invasive examination, can repeatedly detect, and improve the nutrition immunity state of patient by the intervention of specific intestinal flora.This provides a new way for personalized medicine, helps doctor to formulate more accurate treatment strategy.

The invention discloses a method for increasing the viable count of clostridium butyricum through fermentation-separation coupling, which adopts anion exchange resin to perform in-situ separation on organic acid generated in the fermentation process so as to increase the viable count of clostridium butyricum. The clostridium butyricum is cultured in a constant-pH (7.0) fed-batch fermentation mode, and when the glucose content in fermentation liquor is lower than 5 g / L, a certain amount of fresh culture medium is added to enable the glucose concentration to reach 10-15 g / L; when the pH value of the fermentation environment is lower than 6.5, a proper amount of anion exchange resin is added into the culture medium, and the pH value of the fermentation liquid is adjusted to 7.0. Ion exchange resin is used for establishing a fermentation-separation coupling process, growth inhibition of organic acid on clostridium butyricum in the fermentation process is relieved, the viable count is increased, and a good foundation is laid for the preparation process of the clostridium butyricum microbial inoculum. And the ion exchange resin can be recycled after being eluted, so that the purpose of reducing the cost is achieved.

The present invention discloses a mutant of glutamate dehydrogenase derived from Clostridium symbiosum ( Clostridium symbiosum ) and its application in the synthesis of L - glufosinate. The present invention uses a rational analysis and design mutation modification method for amino acid residues in the substrate - binding pocket of glutamate dehydrogenase, the geometry in the molecular tunnel, and the interaction between domains in the domain, and performs single - site or multi - site mutations on the wild - type glutamate dehydrogenase derived from Clostridium symbiosum shown in SEQ ID NO.1. The obtained single - site or multi - site mutants significantly improve the catalytic activity towards 2 - oxo - 4 - (hydroxymethylphosphinyl) butyric acid and have application prospects in the biocatalytic preparation of L - glufosinate.

The invention discloses an RNA methylation detection kit based on a CRISPR / CtCas12a system and a detection method, and belongs to the technical field of gene detection and molecular diagnosis. According to the invention, heat-resistant Cas12a protein derived from clostridium butyricum is utilized, and an ss-ds DNA activator is rationally designed, so that the difference between trans-cleavage activity of CtCas12a activated by methylated RNA and trans-cleavage activity of CtCas12a activated by unmethylated RNA is remarkably amplified; by further combining with a detection probe, high-sensitivity and single-base-resolution quantitative analysis of methylation modification of multiple RNAs such as m6A and m1A is synchronously realized under the condition that nucleic acid amplification is not needed, and an efficient tool is provided for disease marker screening and clinical precise diagnosis and treatment; besides, the method overcomes antibody dependence and amplification deviation, and is suitable for rapid detection of clinical samples and dynamic monitoring of epitranscriptome, so that the method has a better application prospect in RNA methylation detection.

The invention belongs to the technical field of molecular biology and biology, and particularly relates to application of YnjE protein in preparation of a clostridium perfringens resisting product. The amino acid sequence of the YnjE protein is as shown in SEQ ID NO. 1. The YnjE protein shows a high protection rate of 85.7% and high immunogenicity in immune protection in clostridium perfringens, in addition, as a surface exposed protein coded by a core gene, the YnjE protein is highly conserved in various clostridium perfringens, and due to the characteristic that the YnjE protein is easily recognized by an immune system, the YnjE protein can stimulate effective immune response, so that the immunogenicity of the YnjE protein is improved, and the immunogenicity of the YnjE protein is improved. The polypeptide has the potential of becoming a vaccine candidate.