97 results about "Aminopeptidase" patented technology

The invention relates to the technical field of microorganisms, in particular to an Aspergillus oryzae ZA307 strain and application of the Aspergillus oryzae ZA307 strain. The aspergillus oryzae ZA307 high-yield protease and aminopeptidase provided by the invention cooperate with aspergillus oryzae Huniang 3.042 for starter propagation, so that the activity of the protease, aminopeptidase, saccharifying enzyme and amylase of a starter material can be remarkably improved, and furthermore, the content of ammonia nitrogen and total nitrogen and the variety and content of volatile flavor substances in a product are remarkably improved. The soy sauce or soybean paste prepared and fermented by double-strain starter propagation is obviously improved in the aspects of sauce fragrance, ester fragrance, delicate flavor and harmonious taste, and the quality of the product is comprehensively improved.

The invention provides collagen tripeptide liquid with moisturizing and anti-photoaging effects and a preparation method thereof, and relates to the technical field of bioactive peptides.The preparation method comprises the steps that pretreatment and first-time enzymolysis are conducted, specifically, pretreated fish skin and water are mixed, the pH value is adjusted, compound protease is added for enzymolysis, and first enzymatic hydrolysate is obtained; second-time enzymolysis: adjusting the pH value of the first enzymatic hydrolysate to 6.0-7.5, adding neutral protease for enzymolysis, and then adding aminopeptidase for enzymolysis for 20-40 minutes to obtain second enzymatic hydrolysate; and performing enzyme deactivation, decoloration, separation and purification to obtain collagen tripeptide liquid. Wherein the content of collagen tripeptide, the content of glycine-proline-hydroxyproline and the content of hydroxyproline are high, and good moisturizing and anti-photoaging effects are achieved.

The invention relates to the technical field of polypeptide biosynthesis, in particular to a method for biosynthesizing acetyl tetrapeptide-5, a composition containing the acetyl tetrapeptide-5 and application of the composition. According to the method, low-cost natural amino acid is used as a substrate, specific aminopeptidase or a mutant thereof is used as a biocatalyst, amino acid ligase and polyphosphatase are combined, and acetyl tetrapeptide-5 can be efficiently synthesized through a one-step enzymatic reaction. Compared with a traditional chemical synthesis method, the method has the advantages of low raw material cost, no use of an organic solvent, mild reaction conditions, high regioselectivity, environmental friendliness, low production cost and the like. The invention also provides a composition containing acetyl tetrapeptide-5 and dipeptide-15, the inhibition effect of the composition on grease secretion is obviously better than that of a single component, and the composition has a synergistic oil control effect under a specific ratio.

The invention relates to a method for reducing the anaphylaxis of natural latex by multi-enzyme cascade catalysis of rubber protein. The invention relates to the technical field of catalysis of rubber protein hydrolysis, in particular to a method for reducing the anaphylaxis of natural latex by thoroughly hydrolyzing rubber protein through multi-enzyme cascade catalysis. The method for reducing the anaphylaxis of the natural latex by catalyzing the rubber protein through the synergistic effect of multiple enzymes from the perspective of industrial production comprises the following steps: adding an enzyme A and an enzyme B into the natural latex for reaction so as to reduce the anaphylaxis of the natural latex, the enzyme A is alkaline protease; and the B enzyme is at least one of aminopeptidase, carboxypeptidase B, carboxypeptidase A and carboxypeptidase Y. According to the method disclosed by the invention, the enzyme A-endopeptidase and the enzyme B-exopeptidase are subjected to multi-enzyme-stage combined treatment and concerted catalysis, so that the enzyme A-endopeptidase and the enzyme B-exopeptidase are completely hydrolyzed or are completely hydrolyzed into more amino acid substances and short-chain peptides (oligopeptides) as far as possible, and the amino acid substances and the short-chain peptides (oligopeptides) are retained in latex. Protein is highly hydrolyzed into amino acid with higher safety, so that the possibility of I-type anaphylactic reaction caused by protein allergens is greatly reduced or even eliminated.

The invention belongs to the technical field of food, and provides a special enzyme composition for infant soybean milk powder and soybean milk powder thereof. A three-stage directional enzymolysis technology is adopted, the three-stage directional enzymolysis technology comprises glutamine transaminase deamidation pretreatment, bacillus subtilis protease-trypsin-papain first-stage hydrolysis and aminopeptidase-phytase-beta-glucosidase second-stage hydrolysis, and conductivity control, carbon dioxide ventilation in-situ complexation and spray drying processes are combined. The free amino nitrogen of the product is increased by more than or equal to 20%, the proportion of 200-1500Da peptide fragments is more than or equal to 70%, the median particle size is 50-180 microns, the water activity is less than or equal to 0.30, the instant dissolving time is less than or equal to 30 seconds at 25 DEG C under the water hardness of 15-200ppm, the settling rate is less than or equal to 3% after standing for 30 minutes, and the zeta potential is-10 to-25mV. The palatability contradiction between deep hydrolysis and bitterness inhibition of the bean-based infant formula powder, the brewing contradiction between high-calcium reinforcement and instant stability and the process controllability contradiction between three-stage enzymolysis and ionic strength accumulation are effectively solved, and the bean-based infant formula powder has wide application value in the fields of bean-based infant formula food and vegetable protein nutritional products.

The invention relates to a composition for bone repair and growth and a preparation method thereof, and belongs to the technical field of medicines, the composition comprises a glue layer extracted peptide in sandjellyfish and a tabasheer-grifola frondosa composite bacterial peptide; the extract peptide of the glue layer in the sand jellyfish contains a product between 500Da and 2000Da, wherein the product is obtained by sequentially carrying out reaction on the glue layer in the sand jellyfish through lumbrukinase, serrapeptase and glutamine transaminase; the tabasheer-grifola frondosa composite bacterial peptide contains a product between 500Da and 2000Da, which is obtained by performing common enzymolysis on tabasheer mushrooms and grifola frondosa mushrooms through cellulase and chitinase, then performing common enzymolysis on the tabasheer mushrooms and the grifola frondosa mushrooms through nattokinase and serrapeptase and finally performing common reaction on aminopeptidase and glutamine transaminase. According to the invention, a special method is adopted to prepare the glue layer extracted peptide and the tabasheer-grifola frondosa compound bacterium peptide in the sandfish, and the glue layer extracted peptide and the tabasheer-grifola frondosa compound bacterium peptide are matched for use to achieve good biological activity, stability and absorption and utilization rate, so that the defects of the existing skeleton repair and growth medicines are overcome, and the repair and growth of skeletons are effectively promoted.

The invention provides a method for efficiently preparing active polypeptide by fermenting rapeseed dregs, which comprises the following steps: pretreatment: crushing rapeseed dregs, adding 3-5% of reducing sugar, humidifying and sterilizing; performing mixed bacteria segmented fermentation: inoculating a composite strain consisting of bacillus licheniformis SWJS33 with high yield of aminopeptidase, aspergillus oryzae and lactobacillus plantarum, and fermenting at 30 DEG C or 37 DEG C for 72-84 hours; ultrasonic wall breaking: adding water after fermentation, uniformly mixing, carrying out ultrasonic treatment for 20-30 minutes under the power of 120-200W, and breaking thalli to release intracellular enzyme; enzymolysis: carrying out enzymolysis for 4-6 hours under the condition of 50-60 DEG C; post-treatment: enzyme deactivation, centrifugation and drying are carried out after enzymolysis, and a polypeptide product is obtained. By adding the exogenous reducing sugar, nutrition is enhanced, and thallus proliferation is accelerated; bacillus licheniformis SWJS33 with high yield of aminopeptidase is inoculated, so that the generation of small-molecule active peptide is further promoted; a mixed bacteria composite system and a segmented fermentation technology are adopted, and bacteria cells are crushed in an ultrasonic-assisted manner, so that the enzymolysis efficiency is enhanced, and the yield and activity of the polypeptide are remarkably improved.

The present disclosure provides compounds of Formula (I), which have activity as inhibitors of aminopeptidase enzymes. The present disclosure provides methods and uses of the compounds in the treatment of diseases, particularly conditions and diseases associated with aminopeptidase.

The present invention belongs to the field of genetic engineering and enzyme engineering, and relates to a chassis strain lacking the bglS gene and its application. The present invention provides a genetically engineered chassis strain of Bacillus subtilis, wherein the chassis strain is constructed by deleting the glucanase encoding gene on the Bacillus subtilis host genome. bglS The described chassis bacteria can efficiently express alkaline protease, aminopeptidase, mesophilic α-amylase, and thermostable α-amylase. This method is also suitable for increasing the expression of other exocrine proteins, laying the foundation for efficient expression of heterologous proteins and promoting the efficient expression and industrial production of alkaline protease, aminopeptidase, mesophilic α-amylase, and thermostable α-amylase.

The invention relates to the field of enzyme engineering, and discloses an aminopeptidase mutant as well as a coding gene, a recombinant vector, a recombinant strain and application thereof, and the aminopeptidase mutant is obtained by performing mutation of at least one of S88L, Y150C and Q268F on an enzyme with an amino acid sequence as shown in SEQ ID NO: 1. The aminopeptidase mutant disclosed by the invention can catalyze L-histidine to synthesize L-carnosine in a high-selectivity manner in the presence of beta-alanine methyl ester, and the aminopeptidase mutant has relatively high L-carnosine yield and relatively low impurity content, and has industrial application value.

The present invention relates to a composition for bone repair and growth and a preparation method thereof, belonging to the field of medical technology. The composition includes a peptide extracted from the gelatinous layer of sand jellyfish and a bamboo yellow-Grifola frondosa composite bacterial peptide; the peptide extracted from the gelatinous layer of sand jellyfish contains a product between 500Da and 2000Da obtained by sequentially hydrolyzing the gelatinous layer of sand jellyfish with lumbrokinase, serrapeptase, and transglutaminase; the bamboo yellow-Grifola frondosa composite bacterial peptide contains a product between 500Da and 2000Da obtained by co-enzymatic hydrolysis of bamboo yellow mushrooms and Grifola frondosa mushrooms with cellulase and chitinase, then co-enzymatic hydrolysis with nattokinase and serrapeptase, and finally co-enzymatic hydrolysis with aminopeptidase and transglutaminase. The present invention adopts a special method to prepare the peptide extracted from the gelatinous layer of sand jellyfish and the bamboo yellow-Grifola frondosa composite bacterial peptide, and the combined use achieves good biological activity, stability, and absorption utilization rate, overcomes the defects of existing bone repair and growth drugs, and effectively promotes bone repair and growth.

The invention belongs to the technical field of medicines, and discloses a near-infrared fluorescent probe Cy-NEO-Leu for leucine aminopeptidase responsiveness as well as a preparation method and application of the near-infrared fluorescent probe Cy-NEO-Leu. The parent nucleus of the probe is of a heptamethyl cyanine structure, and the probe has high molar absorption coefficient, high fluorescence quantum yield and excellent phototherapy characteristics. The introduction of a neomycin group can increase the water solubility of the probe, and a neomycin multi-ammoniation structure exists in a polycation form under physiological conditions and can target negatively charged cell walls of gram-negative bacteria, so that the probe Cy-NEO-Leu is enriched around the bacteria, can be specifically recognized by leucine aminopeptidase and can be hydrolyzed and activated; and photo-thermal and photodynamic sterilization effects are achieved under laser irradiation. Due to the good biocompatibility, the remarkable active oxygen generation capacity, the excellent photo-thermal conversion efficiency and the unique property of accurately targeting bacteria, the Cy-NEO-Leu is expected to play an important role in a photo-thermal / photodynamic combined treatment scheme, and is used for resisting diseases caused by bacteria and biological membranes and accelerating the healing process of wounds.

The invention discloses an LAT1-mediated STING agonist prodrug as well as a preparation method and medical application thereof, and belongs to the technical field of medicines. In order to solve the problem of specific delivery of the STING agonist, a series of LAT1-mediated STING agonist prodrug molecules are designed and synthesized, the molecules enter tissues through recognition and transportation of LAT1, then the STING agonist molecules are subjected to hydrolysis bond breaking through esterase or aminopeptidase, and the STING agonist molecules are released, so that the STING-activated non-specific immunocompetence is realized.

Disclosed are a specific marker combination of mesenchymal stem cell-derived small extracellular vesicles and the use thereof. The characteristic marker combination comprises aminopeptidase N (CD13), integrin β-1 (CD29), membrane glycoprotein THY-1 (CD90), and markers CD9, CD63 and CD81. By using the characteristic marker combination of mesenchymal stem cell-derived small extracellular vesicles for nano-flow cytometry analysis at the single-vesicle level, mesenchymal stem cell-derived small extracellular vesicles can be identified. On the basis of the provided characteristic marker combination, specific modification and engineering on the basis of small extracellular vesicle membrane proteins can further be realized.

The application belongs to the technical field of fluorescent probe, and particularly relates to an aminopeptidase fluorescent probe and a preparation method and application thereof. In order to develop an APN near-infrared fluorescent probe suitable for in vivo imaging, a center-position ester group substituted pentamethine cyanine dye platform (Cy5-COOM) is used, and based on an "ester group to carboxyl group" conversion strategy, an aminopeptidase (APN) fluorescent probe Cy5-APN is developed. The probe is firstly hydrolyzed by APN, then a 1,6-elimination reaction occurs, and finally a strong fluorescent center-position carboxyl group substituted pentamethine cyanine dye Cy5-COO is released. In addition, in view of the characteristics that cancer cell surfaces overexpress APN, the probe can distinguish cancer cells / tissues and normal cells / tissues with high contrast, and therefore has great application potential in fluorescence-guided tumor surgery.

The present application relates to a kind of by knocking out Bacillus subtilis SacB gene constructed chassis strain and its application, belong to genetic engineering, enzyme engineering and food engineering technical field.The present application is by SacB gene on the genome of Bacillus subtilis Knocking out, obtain a kind of can efficiently express aminopeptidase, alkaline protease Bacillus subtilis expression system, lay the foundation for the efficient expression of protein, promote the efficient expression and industrial production of aminopeptidase, alkaline protease.The chassis strain described above, in the case of carrying aminopeptidase, alkaline protease expression cassette, respectively, 48H shake flask fermentation, fermentation broth aminopeptidase enzyme activity is as high as 1666.7U / mL, with the enzyme activity of starting strain as contrast, enzyme activity is increased by about 30%;Fermentation broth alkaline protease enzyme activity is as high as 6348U / mL, with the enzyme activity of starting strain as contrast, enzyme activity is increased by about 20%.

The invention belongs to the technical field of pet food, and particularly relates to a protein cat food flavoring agent and a preparation method thereof.The preparation method of the protein cat food flavoring agent comprises the steps that meat extract, silkworm chrysalis homogenate, euphausia superba homogenate, pollen pini, ethyl maltol, a flavoring agent, trypsin, neutral protease and aminopeptidase are sequentially added for enzymolysis; an enzymolysis product is obtained; adding goat milk powder, carbohydrates, microencapsulated thiamine, pumpkin powder and amino acid into the reaction tank, uniformly mixing with the enzymolysis product, and carrying out Maillard reaction; after the Maillard reaction is finished, phosphoric acid and a thickening agent are added into the reaction tank, and a pasty flavoring agent is obtained; and carrying out spray drying on the pasty flavoring agent to obtain a powdery flavoring agent. The components jointly act on the cat food flavoring agent, so that the nutritional value of the flavoring agent is improved, the flavor level of the flavoring agent is richer and diversified, the flavoring agent can meet the favor of cats for flavor, and meanwhile, comprehensive nutritional support is provided.

The invention provides a fluorescent probe compound which is novel in structure and shown in a formula (I) and application of the fluorescent probe compound in detection of aminopeptidase N. The fluorescent probe compound can detect the aminopeptidase N in an aqueous solution system, has specificity, excellent selectivity, high sensitivity and the like on detection of the aminopeptidase N and can be used for quantitative detection of the aminopeptidase N. The preparation method of the fluorescent probe compound as shown in the formula (I) is simple in synthesis step, good in biocompatibility, short in reaction time, convenient to purify and simple in process.

This invention provides a feed additive and compound feed for preventing and treating coccidiosis in broilers, comprising the following components: *Kluyveromyces martensii*, ferulic acid esterase, aminopeptidase, and glucanase. Preferably, the mass percentage of *Kluyveromyces martensii* is 30-40%; the mass percentage of ferulic acid esterase is 15-28%; the mass percentage of aminopeptidase is 15-25%; and the mass percentage of glucanase is 18-25%. Based on extensive research, this invention has determined that this combination has a significant effect on coccidiosis control in broilers, effectively improving broiler production performance indicators, increasing economic benefits, and reducing the use of anticoccidial drugs.

The invention discloses a fluorescent probe for detecting arginine aminopeptidase as well as a preparation method and application of the fluorescent probe, and belongs to the technical field of biological medicines. The fluorescent probe (DDAR) provided by the invention can be hydrolyzed by arginine aminopeptidase (RAP) at high selectivity to generate a hydrolysate of which the fluorescence attribute is remarkably changed, so that high-specificity detection can be realized through a fluorescence detector. Moreover, the probe reaction system can be used for quantitatively evaluating the activity and function of the arginine aminopeptidase in an intestinal microorganism sample, and can be used for screening an arginine aminopeptidase inhibitor. The DDAR provided by the invention has long fluorescence emission wavelength, can well weaken interference of biological background fluorescence, and has high sensitivity. The fluorescent probe for detecting arginine aminopeptidase provided by the invention can be prepared through a simple chemical synthesis process, the preparation process is simple and easy to implement, the detection cost of a fluorescent method is low, and the fluorescent probe has a very good application value.

The present invention discloses a fluorescent probe based on a silicon-substituted rhodamine derivative, its preparation method, and its application, relating to the field of biochemistry. The present invention provides the structural formula of the fluorescent probe based on the silicon-substituted rhodamine derivative and also provides a corresponding preparation method. The present invention designs a leucine aminopeptidase-responsive near-infrared fluorescent small molecule probe that can effectively identify head and neck squamous cell carcinoma, effectively addressing the low sensitivity, specificity, and response speed issues of existing technologies.

The present application provides compositions and methods for determining a disease or condition in a subject. The method comprises contacting a body fluid with a molecule comprising a reporter thereof and the reported is cleaved by an agent in the body fluid. Diseases and conditions that can be determined by the methods are also described herein

The application provides a near-infrared fluorescent probe based on basic dye and a preparation method and application thereof, and the probe is named 4-(2-amino-4-methylpentanoylamino)benzyl 3,7-bis(diethylamino)-10H-phenoxazine-10-methylate in Chinese. The probe is used for detecting leucine aminopeptidase (LAP), basic blue 3 is used as a fluorophore, and L-leucine is used as a recognition site of the LAP. When the probe is reacted with the LAP, the leucine is hydrolyzed, the basic blue 3 fluorophore is released through a self-immolative group, red fluorescence is emitted, the probe can be used for detecting the leucine aminopeptidase, the detection signal is obvious, the color change of a reaction solution is visible to the naked eye, and the probe provides an effective visual fluorescent tracking tool for the leucine aminopeptidase for biological detection.

A composition for use in treating or preventing colorectal cancer (CRC) comprising indole-3-Iactic acid (ILA) or an aminopeptidase produced by Lactococcus lactis, the aminopeptidase having a molecular

The invention discloses a directional preparation method of a coprinus comatus protein peptide Maillard reaction product with salt reducing and saltiness increasing capabilities. The directional preparation method comprises the following steps: S1, pretreatment; s2, raw material endogenous enzyme inactivation; s3, first-step enzymolysis: adding cellulase for enzymolysis; s4, performing enzyme deactivation treatment; s5, second-step enzymolysis: adding flavourzyme and aminopeptidase for enzymolysis; s6, performing enzyme deactivation treatment; s7, preparing supernate of the enzymatic hydrolysate; s8, preparation of a Maillard reaction product: adding D-xylose and L-cysteine to carry out a Maillard thermal reaction, and obtaining the coprinus comatus protein peptide Maillard reaction product after the reaction; wherein the addition amount of the D-xylose is 10-12% of the solid content of the enzymatic hydrolysate, and the addition amount of the L-cysteine is 6-13% of the solid content of the enzymatic hydrolysate. According to the method, cellulase, flavourzyme and aminopeptidase are selected to perform two-step hydrolysis on the coprinus comatus, then the Maillard reaction is performed, the ratio of D-xylose to L-cysteine and the reaction time are regulated and controlled, freshness and saltiness are increased to the maximum extent, and the fragrance or meat fragrance of the coprinus comatus is reserved.

The present invention belongs to the field of synthetic biology, and specifically relates to a kind of L-carnosine and its preparation method and aminopeptidase. The present invention is based on the theory of synthetic biology and develops a preparation method of L-carnosine based on homemade aminopeptidase and produces high-purity L-carnosine. The preparation method of L-carnosine of the present invention can achieve the recovery and reuse of L-histidine while obtaining high-purity L-carnosine, effectively reducing costs. In addition, the preparation method of the present invention has mild reaction conditions, a short synthetic route, is environmentally friendly, and the product has high optical purity.

The invention belongs to the technical field of protein peptide powder. The invention provides a preparation method of ovalbumin peptide powder, which comprises the following steps: S1, transferring protein liquid into a fermentation tank, adjusting the pH value to 6.0-6.9, adding dry yeast, stirring and fermenting to obtain fermentation liquid; s2, transferring the fermentation liquor obtained in the S1 into an enzymolysis tank, heating to 50-55 DEG C, adding a compound enzyme preparation, carrying out enzymolysis for 7-10 hours, and stirring for 10 minutes per hour to obtain enzymatic hydrolysate; the compound enzyme preparation comprises papain, subtilisin, trypsin and aminopeptidase; and S3, transferring the enzymatic hydrolysate obtained in S2 back to a fermentation tank, cooling, adding dry yeast, stirring, fermenting, spray-drying, and placing in a hot chamber for 3-7 days to obtain the product. According to the ovalbumin peptide powder, the oxidation resistance and the heat resistance are improved at the same time, so that the ovalbumin peptide powder with more excellent functionality and wider application range is obtained.

The invention belongs to the field of waste resource utilization, and relates to a method for preparing and producing a liquid amino acid-peptide water-soluble fertilizer by utilizing waste animal protein. Neutral protease and alkaline protease are adopted for compound enzymolysis of animal protein, the protein transfer rate is greatly increased, and the protein transfer rate reaches the maximum value of 76.42%. Then, aminopeptidase is utilized for deep hydrolysis, and finally, inorganic ions are chelated to prepare the amino acid-peptide water-soluble fertilizer. The content of dissolved peptide in the amino acid-peptide water-soluble fertilizer prepared by the invention can reach 116.26 g / L, and the total amount of free amino acid can reach 8235 mg / L. The proportion of small molecule peptide and amino acid is high, and the proportion of oligopeptide with the molecular weight of 180-500 Da can reach 62.1%; and the proportion of components less than 180Da can reach 12.9%. The water-soluble fertilizer prepared by the invention can promote the growth of spinach and enhance the enzyme activity of soil, and is more beneficial to the best balance of spinach absorption and soil improvement compared with a long-peptide peptide fertilizer.

The invention relates to an aminopeptidase gene derived from aspergillus oryzae and an amino acid sequence of the aminopeptidase gene. Through optimization design, the expression efficiency of the gene in pichia pastoris is remarkably improved. The invention also provides an expression vector containing the gene and a genetic engineering recombinant strain carrying the vector. By adding a specific leader peptide sequence at the front end of a wild type aminopeptidase gene, high-efficiency expression of aminopeptidase in pichia pastoris is realized, and the yield of the aminopeptidase is greatly increased, so that possibility is provided for large-scale industrial production, and the production cost is remarkably reduced. The recombinant strain can secrete extracellular soluble aminopeptidase, the subsequent extraction and purification process is simplified, and industrial application is facilitated. Compared with the existing aminopeptidase in the market, the recombinant aminopeptidase disclosed by the invention has more efficient bitterness removing performance, can achieve an expected effect by only needing a half of addition amount under the same hydrolysis degree, and shows more excellent industrial application potential. The technology provided by the invention breaks through and fills the technical blank in the aspects of efficient expression and industrial production of aminopeptidase in China, and has important economic value and market prospect.

The present invention relates to a hydrolyzed collagen formulation for use in decreasing blood glucose. The hydrolyzed collagen is obtained by enzymatic hydrolysis of a collagen-containing material with a combination of enzymes comprising two or more enzymes selected from the group consisting of a neutral protease, a carboxypeptidase, and an aminopeptidase. The hydrolyzed collagen formulation is particularly suitable as a food supplement, such as for use in ameliorating hyperglycemia and / or a risk factor hyperglycemia.