221 results about "Subtilisin" patented technology

Antagonists of human proprotein convertase subtilisin-kexin type 9 (“PCSK9”) are disclosed. The disclosed antagonists are effective in the inhibition of PCSK9 function and, accordingly, present desirable antagonists for use in the treatment of conditions associated with PCSK9 activity. The present invention also discloses nucleic acid encoding said antagonists, vectors, host cells, and compositions comprising the antagonists. Methods of making PCSK9-specific antagonists as well as methods of using the antagonists for inhibiting or antagonizing PCSK9 function are also disclosed and form important additional aspects of the present disclosure.

New subtilisin homologues (both nucleic acids and proteins) are provided. Compositions which include these new proteins, recombinant cells, shuffling methods involving the new homologues, antibodies to the new homologues, and methods of using the homologues are also provided.

Antagonists of human proprotein convertase subtilisin-kexin type 9 (“PCSK9”) are disclosed. The disclosed antagonists are effective in the inhibition of PCSK9 function and, accordingly, present desirable antagonists for use in the treatment of conditions associated with PCSK9 activity. The present invention also discloses nucleic acid encoding said antagonists, vectors, host cells, and compositions comprising the antagonists. Methods of making PCSK9-specific antagonists as well as methods of using the antagonists for inhibiting or antagonizing PCSK9 function are also disclosed and form important additional aspects of the present disclosure.

The present invention provides antibody antagonists against proprotein convertase subtilisin / kexin type 9a (“PCSK9”) and methods of using such antibodies.

Antagonists of human proprotein convertase subtilisin-kexin type 9 (“PCSK9”) are disclosed. The disclosed antagonists are effective in the inhibition of PCSK9 function and, accordingly, present desirable antagonists for use in the treatment of conditions associated with PCSK9 activity. The present invention also discloses nucleic acid encoding said antagonists, vectors, host cells, and compositions comprising the antagonists. Methods of making PCSK9-specific antagonists as well as methods of using the antagonists for inhibiting or antagonizing PCSK9 function are also disclosed and form important additional aspects of the present disclosure.

The present invention provides antibodies and antigen-binding fragments thereof that specifically bind proprotein convertase subtilisin / kexin-9 (PCSK9) with greater affinity at neutral pH than at acidic pH. The antibodies of the invention may possess one or more amino acid changes as compared to antibodies that do not exhibit pH-dependent binding properties. For example, the present invention includes anti-PCSK9 antibodies which possess one or more histidine substitutions in one or more complementarity determining regions. The antibodies of the invention, with pH-dependent binding properties, remain in circulation and exhibit cholesterol lowering activity for prolonged periods of time in animal subjects as compared to anti-PCSK9 antibodies that do not exhibit pH-dependent binding properties. The antibodies of the invention are therefore useful for treating diseases and disorders related to elevated HDL cholesterol, wherein the antibodies of the invention can be administered to a patient at a lower dose and / or with less frequent dosing as compared to antibodies that do not exhibit pH-dependent binding properties.

An antifouling paint composition comprising an enzyme, such as endopeptidase, Subtilisin (EC 3.4.21.62) and Alcalase(R), and a rosin compound, wherein the enzyme is effective to reduce or prevent fouling by aquatic organisms of a surface coated with the composition. Also disclosed is a method for preventing fouling of a surface by aquatic organisms.

The invention discloses a recombinant bacillus subtilis and a method for producing transglutaminase by utilizing the recombinant bacillus substilis. The recombinant bacillus subtilis is obtained by the following steps of: (1) by virtue of fusion PCR (polymerase chain reaction), connecting a P43 promoter and a signal peptide gene with a subtilisin-like protease gene, and converting a fusion PCR amplification product into bacillus subtilis WB800 to obtain bacillus subtilis B.subtilis WB800S; and (2) carrying out PCR amplification on streptoverticillium mobaraense transglutaminase protogene, andthen converting amplification product into the bacillus subtilis B.subtilis WB800S to obtain bacillus subtilis B.subtilis WB800S / proMTG. The bacillus subtilis B.subtilis WB800S / proMTG is fermented and purified to obtain soluble transglutaminase. By adopting the method disclosed by the invention, the transglutaminase with biological activity can be directly obtained in supernatant of fermentation liquor, fussy processes of breaking cells and activating proenzyme after purification are eliminated, and a fermentation production process is simplified.

In a liquid detergent comprising a subtilisin and optionally a second (non-subtilisin) enzyme, the combination of a peptide aldehyde (or hydrosulfite adduct thereof) with a salt of a monovalent cation and a monovalent organic anion has a synergistic stabilizing effect on the subtilisin and / or the second enzyme. The improved enzyme stability is of particular interest in liquid detergent compositions where the enzyme would otherwise have poor storage stability.

Acid-stable proteases of the subtilisin family, their use in animal feed, feed-additives and feed compositions containing such proteases, and methods for the treatment of vegetable proteins using such proteases.

The present invention provides a Bacillus sp. subtilisin variant. In addition, the present invention provides automatic dishwashing compositions comprising this serine protease variant.

The present invention relates to cleaning compositions comprising a protease variant. One cleaning composition comprises a protease variant including a substitution of an amino acid residue with another naturally occurring amino acid residue at an amino acid residue position corresponding to position 103 of Bacillus amyloliquefaciens subtilisin in combination with a substitution of an amino acid residue with another naturally occurring amino acid residue at one or more amino acid residue positions corresponding to positions 1, 3, 4, 8, 9, 10, 12, 13, 16, 17, 18, 19, 20, 21, 22, 24, 27, 33, 37, 38, 42, 43, 48, 55, 57, 58, 61, 62, 68, 72, 75, 76, 77, 78, 79, 86, 87, 89, 97, 98, 99, 101, 102, 104, 106, 107, 109, 111, 114, 116, 117, 119, 121, 123, 126, 128, 130, 131, 133, 134, 137, 140, 141, 142, 146, 147, 158, 159, 160, 166, 167, 170, 173, 174, 177, 181, 182, 183, 184, 185, 188, 192, 194, 198, 203, 204, 205, 206, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 222, 224, 227, 228, 230, 232, 236, 237, 238, 240, 242, 243, 244, 245, 246, 247, 248, 249, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 265, 268, 269, 270, 271, 272, 274 and 275 of Bacillus amyloliquefaciens subtilisin; wherein when said protease variant includes a substitution of amino acid residues at positions corresponding to positions 103 and 76, there is also a subtitution of an amino acid residue at one or more amino acid residue positions other than amino acid residue positions corresponding to positions 27, 99, 101, 104, 107, 109, 123, 128, 166, 204, 206, 210, 216, 217, 218, 222, 260, 265 or 274 of Bacillus amyloliquefaciens subtilisin; and one or more cleaning adjunct materials. Another cleaning composition comprises a protease variant including a substitution of an amino acid residue with another naturally occurring amino acid residue at one or more amino acid residue positions corresponding to positions 62, 212, 230, 232, 252 and 257 of Bacillus amyloliquefaciens subtilisin; and one or more cleaning adjunct materials. Methods for using the cleaning compositions are also provided.

The invention provides a method for preparing tobacco extract by utilizing waste and inferior tobacco leaves. The tobacco extract is prepared through the following steps: mixing tobacco powder, tobacco leaf fragments and tobacco stems in a cigarette manufacturing department and adding the mixture into an ethanol solution for extracting; decompressing and concentrating; then adding pure water and glucose in sequence; after uniformly stirring, adding cellulase and subtilisin in sequence to carry out enzymolysis; then adding the mixture subject to enzymolysis into the ethanol solution and standing; filtering with a ceramic film; and decompressing and concentrating and then carrying out molecular distillation and separation. According to the method, efficient, rapid, energy-saving and environment-friendly effects are achieved; prepared molecular distillate has a real aroma and the taste is remarkably improved; the product is used as essence and spice for cigarettes so that the aroma of the cigarettes can be obviously improved; and meanwhile, the product also has soft, fine and smooth smoke and has the effects of improving the sweet feeling of the oral cavity and improving the smoke volatility.

Novel calcium free subtilisin mutants are taught, in particular subtilisins which have been mutated to eliminate amino acids 75-83 and which retain enzymatic activity and stability. Recombinant methods for producing same and recombinant DNA encoding for such subtilisin mutants are also provided.

A lotus leaf dye is extracted from lotus leaves. Its extracting process is also disclosed. A method for using said lotus leaf dye to dye wool or wool fabric includes ecological pretreating, modifying wool by the composite enzyme consisting of subtilisin and papain, and mordant dyeing. Its advantage is high dyeing effect.

The invention relates to a preparation method and application of an electrochemical immunosensor for a biomarker, namely a recombinant proprotein convertase subtilisin kexin type 9 (pcsk9) protein for predicting and diagnosing cardiovascular diseases, and belongs to the technical field of electrochemical detection. The preparation method is characterized by comprising the following steps: firstly, performing nitrogen doping on a graphene nanobelt (n-gnrs), performing amination on a fullerene-palladium-platinum nanoparticle (n-C60 / pdpt) composite material and a staphylococcus aureus a protein (spa), and performing layer-by-layer self-assembling so as to immobilize a pcsk9 antibody (ab1); mixing the synthesized nanoparticle-poly-methylene blue (pt-pmb) with the pcsk9 antibody (ab2), and preparing a nano beacon, thereby obtaining the electrochemical immunosensor for pcsk9 protein detection. The electrochemical immunosensor has the advantages of being high in sensitivity, good in specificity and rapid and convenient in detection. A novel method for pcsk9 protein detection is provided, and useful information is provided for clinical prediction and diagnosis on cardiovascular diseases.

The present invention relates to a filamentous fungal cell in which an endogenous alkaline protease activity, an endogenous neutral metalloprotease activity and an endogenous serine protease activity of the subtilisin type have been completely or partially inactivated. Particularly the endogenous alkaline protease activity, the endogenous neutral metalloprotease activity and the endogenous serine protease activity of the subtilisin type are encoded by the alp, npI and pepC genes respectively. The filamentous fungal cell is particularly suitable for production of heterologous proteins such as antibodies.

The invention provides a preparation method of cartilage extract containing non-denatured type II collagen. The method comprises the following steps: degreasing, disinfecting, homogenizing, enzymatic hydrolysis, filtration and drying. In the enzymatic hydrolysis step, the pH of a slurry obtained in the homogenizing step is adjusted to 2.5-8.5, an enzyme accounting for 0.001-2%of the weight of cartilages is added, a clear liquid accounting for 1 / 20 to 1 / 500 of the weight of the cartilages and obtained through juicing pawpaw and / or pineapples and filtering the obtained juice is added, the enzymatic hydrolysis is carried out at 25-50 DEG C for 12-48 h, and the enzyme is preferably selected from one or more of pepsin, subtilisin, alkaline proteases and metalloproteases. The cartilage extract obtained through the preparation method has the advantages of high purity, high extraction rate, and easiness in large-scale production, and richness in non-denatured type II collagen and chondroitin sulfate.

The extraction process of comb shell polysaccharide includes the technological steps of material treatment, homogenizing, ultrasonic treatment, enzymolysis, separating, concentration, alcohol precipitation and drying. In the material treatment, the comb shell meat tissue including leftover is taken and may be stoved or freeze dried; and in the enzymolysis, one or two of trypsin, pepsin and neutral subtilisin are used. The present invention provides the technological path of extracting comb shell polysaccharide and the comb shell polysaccharide product has high purity and high yield, and may be used in developing various functional foods.

Novel subtilases having an improved wash performance on egg stains are disclosed. These subtilases are useful exhibiting excellent or improved wash performance on egg stains when used in e.g. cleaning or detergent compositions, such as laundry detergent compositions and dish wash compositions, including automatic dish wash compositions.

The invention discloses a high-protein growth-promotion crab feed, which is prepared from the raw materials in parts by weight: 5 to 15 parts of aquatic plant, 2 to 6 parts of alga, 3 to 9 parts of kelp powder, 12 to 24 parts of soybean meal, 3 to 9 parts of dried potato powder, 5 to 15 parts of corn flour, 4 to 12 parts of field-snail-meat, 5 to 9 parts of fish meal, 3 to 6 parts of silkworm chrysalis meal, 2 to 4 parts of casein, 3 to 6 parts of vital gluten, 1 to 6 parts of alpha-starch, 2 to 5 parts of fish oil, 4 to 6 parts of soybean oil, 2 to 9 parts of cellulose powder, 3 to 8 parts of amino acid mixture, 4 to 9 parts of vitamin mixture, 1 to 6 parts of mineral mixture, 2 to 9 parts of Chinese medicine additive, 1 to 4 parts of compound feed additive, 2 to 5 parts of subtilisin, 1 to 4 parts of bioactive peptide and 2 to 6 parts of phytase. The crab feed disclosed by the invention has high protein content, can meet the nutritional requirement of each stage of crab growth, is low in cost and comprehensive in nutrition, and can effectively promote growth of crabs and improve yield and quality of the crabs.

The present invention relates to methods of producing parental TY145 subtilase variants and parental BPN' subtilase variants, and to TY145 and BPN' variants having altered properties compared to the parental TY145 / BPN' subtilase variants.

The invention discloses an anti-human PCSK9 (pro-protein convertase subtilisin / kexin 9) chimeric antibody, and preparation and application thereof. The preparation method comprises the following steps: respectively amplifying mouse light chain and heavy chain variable region genes from mouse hybridoma cells, respectively carrying out chimerism with light chain and heavy chain constant region genes of human IgG, and carrying out recombination expression to obtain the human-mouse chimeric antibody. The human-mouse chimeric antibody has favorable affinity with human PCSK9, obviously inhibits the degradation activity of the PCSK9 for liver cell low-density lipoprotein receptors (LDLR), enhances the ingestion of liver cells for LDL-cholesterol (LDL-C), and lowers the cholesterol level in blood.

The invention provides a composite microbial starter, a biologically fermented feed prepared by using the starter, and a preparation method of the feed. Aspergillus niger, yeast and Bacillus subtilisin the composite microbial starter have synergistic effects according to a specific ratio, and enhance the fermentation effect. Fresh citrus residues, rice bran, wheat bran, stone flour and magnesiumsulfate in the biologically fermented feed have a reasonable formula. The biologically fermented feed can provide natural pigments, organic mineral elements, dietary fibers, probiotics and metabolitesthereof, needed by production of high-quality livestock and poultry meat, and has broad application prospects and great market values.

Incorporation of a serine protease inhibitor such as RASI, BASI, WASI (bifunctional alpha-amylase / subtilisin inhibitors of rice, barley and wheat) into a liquid detergent which contains a serine proteasecan stabilize the serine protease and / or a second enzyme and can release the enzyme when the detergent composition is diluted.

A method of generating atelocollagen is disclosed. The method comprises contacting a human telopeptide-comprising collagen with a protease selected from the group consisting of neutrase, subtilisin, ficin recombinant human trypsin and recombinant human pepsin, wherein said human telopeptide-comprising collagen is expressed in a non-animal cell, thereby generating the atelocollagen. Compositions comprising the atelocollagen generated thereby are also disclosed.

A method for treating and / or preventing a proprotein convertase subtilisin / kexin type 9 preproprotein (PCSK9)-susceptible viral infection comprising increasing a PCSK9 activity and / or expression in a biological system infected by the virus, whereby the increased PCSK9 activity and / or expression treats and / or prevents the viral infection in the biological system. Methods of classifying subjects, methods of screening and kits therefore.

The invention discloses a construction method of a gene detection library of familial hypercholesterolemia and a gene detection kit and relates to gene mutation of LDLR (Low-Density Lipoprotein Receptor), APOB (Apolipoprotein B) and PCSK9 (Proprotein Convertase Subtilisin / Kexin type 9). In order to ensure that all target regions (including an entire coding region and an alternative splicing regionof a gene to be detected) are covered and prevent a primer dimer or a short segment from being formed between primers of adjacent amplicons, a PCR (Polymerase Chain Reaction) amplification primer isdivided into two independent primer pools; then primer sequence digestion, sequencing connector connection, library purification and quality detection are carried out; finally, the detection library is constructed. The method has the advantages that library establishment steps are simple and rapid, the cost is effectively reduced, the working amount is reduced, variation types are comprehensive, the detection accuracy reaches 100 percent and the flux is high.