66 results about "Aminopeptidase" patented technology

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 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 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 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 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%.

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 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 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 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.

The invention relates to the technical field of microorganisms, in particular to bacillus velezensis phb05 and application of the bacillus velezensis phb05. The bacillus velezensis phb05 strain provided by the invention has the advantages of salt tolerance and high yield of arginine aminopeptidase and glutamic acid aminopeptidase, and has strong degradation capacity for Arg-pNA and Glu-pNA substrates. The strain can be used for preparing an enzyme preparation, is used for food processing, can achieve multiple effects of improving the protein utilization rate, reducing the formation of bitter taste of food and increasing delicate flavor, can improve the flavor of the food, and has very important application in processing of high-salt food.

The present disclosure provides a therapeutic method of treating cancer metastasis by inducing clearance of EVs using a binding agent specific to an EV protein. The method utilizes one or more binding agents specific to EV proteins, where the EV proteins are selected from prostaglandin F2 receptor negative regulator (PTGFRN); basigin (BSG); immunoglobulin superfamily member 2 (IGSF2); immunoglobulin superfamily member 3 (IGSF3); immunoglobulin superfamily member 8 (IGSF8); integrin beta-1 (ITGB1); integrin alpha-4 (ITGA4); 4F2 cell-surface antigen heavy chain (SLC3A2); a class of ATP transporter proteins (ATP1A1, ATP1A2, ATP1A3, ATP1A4, ATP1B3, ATP2B1, ATP2B2, ATP2B3, ATP2B4); CD13 (aminopeptidase N); MME (neprilysin), ENPP1 (ectonucleotide pyrophosphatase / phosphodiesterase family member 1); and NRP1 (neuropilin-1). Further provided herein includes a pharmaceutical composition for the treatment of cancer metastasis.

The objective is to provide an active ingredient for topical skin preparations or food and beverages that is derived from natural products, has excellent biocompatibility and stability, and further possesses excellent skin penetration and metabolic properties. [Solution] A topical skin preparation or food / beverage containing a fish scale extract solution or a fermented product of fish scales by microorganisms as an active ingredient, characterized in that the microorganisms have protease activity or aminopeptidase activity.

This invention relates to the technical field of deep processing of rice protein, and particularly to a fermented rice peptide with α-amylase inhibitory activity, anti-inflammatory and anti-glycation properties, its preparation method, and functional products. The preparation method includes the following steps: mixing rice protein with water to obtain a slurry; adding alkaline protease to the slurry for a first hydrolysis, followed by enzyme inactivation to obtain a first hydrolysate; adding aminopeptidase to the first hydrolysate for a second hydrolysis, followed by enzyme inactivation to obtain a second hydrolysate; adding *Lactobacillus paracasei* to the second hydrolysate for fermentation to obtain a fermentation broth; separating the fermentation broth into solid and liquid phases to obtain a supernatant containing fermented rice peptides; filtering the supernatant to retain the fermented rice peptides, thus obtaining the final product. This invention not only improves the yield of rice peptides but also significantly increases the production efficiency of rice peptides.

The invention discloses an activated double-locking fluorescent probe as well as a preparation method and application thereof, and relates to the technical field of new material technology and biomedical detection. The activated double-lock fluorescent probe is formed by covalently connecting a fluorescent parent nucleus, an acid-sensitive response unit and an enzyme specific recognition response unit, and precise tumor recognition is realized through a cascade activation mechanism: firstly, cracking an acid-sensitive linking group in a tumor acidic microenvironment to expose an amino acid sequence capable of being specifically recognized by aminopeptidase N; and releasing the fluorescent parent nucleus after enzyme digestion reaction. The double-lock fluorescent probe overcomes the false positive problem caused by response of a single biomarker, and has the remarkable advantages of hypersensitive signal response, rapid dynamic response, multi-modal imaging compatibility, broad-spectrum applicability, specificity and the like.

The application provides a preparation method of small molecule enzyme hydrolysis royal jelly with improved absorption, and belongs to the technical field of royal jelly preparation. The method comprises the following steps: (1) preparing a royal jelly aqueous solution; (2) taking the royal jelly aqueous solution, adjusting the pH value to 7-8, then adding sea cucumber hydrolytic enzyme and aminopeptidase, performing enzyme hydrolysis, sterilizing, and obtaining enzyme hydrolysate 1; (3) taking the enzyme hydrolysate 1, adjusting the pH value to 1-2, adding pepsin, performing enzyme hydrolysis, sterilizing, and freeze-drying to obtain the small molecule enzyme hydrolysis royal jelly. The small molecule enzyme hydrolysis royal jelly prepared by the method has a peptide segment molecular weight less than or equal to 1000 Da, wherein the proportion of the peptide segment molecular weight less than or equal to 500 Da is greater than or equal to 80%, the small molecule enzyme hydrolysis royal jelly is easy to be absorbed and utilized by the human body, and has high bioavailability.

The invention belongs to the technical field of biological prevention and control, and provides a dsRNA nucleic acid pesticide for preventing and controlling cedar macroptera longipes and application of the dsRNA nucleic acid pesticide for preventing and controlling cedar macroptera longipes, and the application is that a dsRNA sequence of Aminopeptidase is used for silencing expression of Aminopeptidase. According to the invention, by synthesizing the dsRNA sequence, the effect of the dsRNA sequence in Aminopeptidase expression and the aphid killing effect are evaluated; the result shows that the dsRNA sequence corresponding to the Aminopeptidase of the cedar aphis longibrachii can effectively inhibit the expression of the Aminopeptidase, so that the cedar aphis longibrachii is killed, and the purpose of pest control is achieved.

The invention belongs to the technical field of microbial fermentation, and particularly provides a method for producing aminopeptidase by aspergillus niger fermentation, which comprises the following steps: (1) activating aspergillus niger, and inoculating into a fermentation medium for fermentation; (2) after the carbon source of the base material is used up and the pH value is rebounded, feeding and supplementing are started, and the thalli are controlled to be in a branched filamentous shape through the supplementing speed; and (3) after the dissolved oxygen rebounds, increasing the material supplementing speed, and controlling the fungus shape to be spherical until the fungus is discharged from the tank. According to the method for producing aminopeptidase through aspergillus niger fermentation, the forms of thalli in different periods are controlled by controlling the feed supplement amount in the enrichment period and the enzyme production period in stages, the thalli are controlled to be branch filamentous in the enrichment period, the thalli are controlled to be spherical in the enzyme production period, and the method is different from conventional aspergillus niger fermentation in which the thalli are in filamentous forms in the whole process; after the aspergillus niger is converted from filamentous to spherical, nutrition utilization, energy distribution and environmental adaptability are optimized, and the enzyme yield is increased.

This disclosure concerns production of product peptides with a target amino acid sequence by proteolysis of a recombinant polypeptide comprising specific protease recognition sites or chemical cleavage sequences. In some embodiments, the product peptide is released from repeating peptide units in the recombinant polypeptide by removal of intervening amino acid sequences by proteolysis by proteases that recognize sites within the intervening amino acid sequences and a carboxypeptidase, aminopeptidase, and / or further protease.

The application discloses a clinical biochemical composite quality control product, which comprises detection item raw materials, a matrix liquid and a protective agent; the detection item raw materials are alkaline phosphatase, amylase, sodium glycocholate, cholinesterase, creatinine, leucine aminopeptidase, lactate dehydrogenase, lipase, triglyceride, cholesterol, uric acid, urea, alanine aminotransferase, aspartate aminotransferase, gamma-glutamyl transferase, glucose, alpha-hydroxybutyric acid dehydrogenase, sodium hydrogen phosphate, magnesium chloride, calcium chloride, ferrous chloride, bilirubin, pancreatic amylase and sodium bicarbonate; the matrix liquid is bovine serum albumin and human serum albumin; and the protective agent is dithiothreitol (DTT), mannitol, trehalose, sodium chloride, a composite enzyme stabilizer AES and sodium azide. The quality control product can realize the detection of 27 clinical biochemical items, and the freeze-drying process is adopted, so that the freeze-dried composite quality control product has low viscosity, is easy to be re-dissolved and mixed, has high stability and has small matrix effect.

The application discloses a flower glue peptide with ovary protection and whitening effects and a preparation method and application thereof. The preparation method comprises the following steps: uniformly mixing flower glue of a sea bass in a certain proportion, homogenizing the flower glue by using a homogenizer, sterilizing the flower glue by using a high-pressure sterilization pot, performing fermentation treatment on the flower glue by using Lactobacillus mucosus JIAN, adjusting the pH of the treated flower glue solution by using hydrochloric acid, adding acid protease of Aspergillus niger for enzyme hydrolysis, and then adding aminopeptidase for enzyme hydrolysis, so as to obtain a flower glue peptide solution. The flower glue peptide prepared by the application has significant ovary protection and whitening functions. The whitening function is mainly embodied in tyrosinase inhibition activity, and the ovary protection function is embodied in hydrogen peroxide-induced ovary cell survival rate and anti-mullerian hormone content, so that the economic value of the sea bass by-product is greatly improved. In addition, the flower glue peptide also has the characteristics of weak fishy smell, low fat content and small molecular weight.

The application relates to the field of enzyme engineering, and discloses an aminopeptidase mutant, a coding gene of the aminopeptidase mutant, a recombination carrier, a recombination strain and application of the aminopeptidase mutant, wherein the aminopeptidase mutant is an enzyme with at least one mutation of S88L, Y150C and Q268F in an amino acid sequence shown in SEQ ID NO:1. The aminopeptidase mutant can catalyze synthesis of L-carnosine from L-histidine in the presence of beta-alanine methyl ester, has high selectivity, high L-carnosine yield and low impurity content, and has the value of industrial application.

This invention provides a mushroom culture medium and its preparation method: A compound biological agent is prepared by mixing a compound enzyme preparation with Bacillus subtilis fermentation waste liquid at a mass ratio of 1:50-1:100 to obtain the biological agent. The compound enzyme preparation includes 50,000-100,000 U / g of protease, 10,000-20,000 U / g of acidic cellulase, 10,000-20,000 U / g of neutral cellulase, 50,000-100,000 U / g of xylanase, laccase, 1,000-5,000 U / g of aminopeptidase, and 10,000-20,000 U / g of glucose oxidase. The durian shells are crushed and mixed with the biological agent at a mass ratio of 3:7-1:1. Solid-state fermentation is then performed on the uniformly mixed durian shells at a fermentation temperature of 30-37℃ for 48-96 hours. After fermentation, the durian shells are used as a mushroom culture medium for mushroom cultivation, thus solving the problem of disposing of durian shells and Bacillus subtilis fermentation waste liquid.