36 results about "Thermolysin" patented technology

A process for inhibiting vascular proliferation separately introduces components into the eye to generate plasmin in the eye in amounts to induce complete posterior vitreous detachment where the vitreoretinal interface is devoid of cortical vitreous remnants. The process administers a combination of lysine-plasminogen, at least one recombinant plasminogen activator selected from the group consisting of urokinase, streptokinase, tissue plasminogen activator, chondroitinase, pro-urokinase, retavase, metaloproteinase, and thermolysin and a gaseous adjuvant to form a cavity in the vitreous. The composition is introduced into the vitreous in an amount effective to induce crosslinking of the vitreous and to induce substantially complete posterior vitreous detachment from the retina without causing inflammation of the retina. The gaseous adjuvant material is introduced into the vitreous simultaneously with or after the lysine-plasminogen and recombinant plasminogen activator such as recombinant urokinase to compress the vitreous against the retina while the composition induces the complete posterior vitreous detachment.

The present invention provides an angiotensin converting enzyme inhibitor having strong activity of inhibiting angiotensin converting enzyme, good hue and easily to take in which is a composition containing peptides obtained by digesting a fish meat with thermolysin enzyme, and wherein a content of a polypeptide ingredient having a molecular weight of at least 5000 is at most 10% by weight.

Disclosed is a hydrogel comprising a hydrophilic gelling agent that includes a nonionic cellulose ether, and active thermolysin, wherein the proteolytic activity of the thermolysin does not decrease by more than 20% when stored at room temperature for 6 months.

The invention discloses a method for preparing active polypeptides from deep-sea fish meat, and the method includes degreasing, first deodorization, enzymolysis, second deodorization, ultrafiltration,decolorization, desalting and freeze-drying. 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide is used to activate chitosan, and coupled with glutaraldehyde to form a lipase immobilized carrier, an active short peptide is used to improve the physical and chemical adsorption force between a carrier and lipase, the lipase stability, activity, heat resistance and use frequency are improved; through useof the complementary effect and synergistic effect of thermolysin and chymotrypsin hydrolysis sites, the protein hydrolysis degree and enzymatic hydrolysis speed of minced deep-sea fish meat can be improved, terminal group type can be changed, and the active polypeptides with antitumour activity can be obtained by hydrolysis.

The present invention provides methods and compositions comprising at least one thermolysin-like neutral protease enzyme with improved storage stability and / or catalytic activity. In some embodiments, the thermolysin finds use in cleaning and other applications comprising detergent. In some particularly preferred embodiments, the present invention provides methods and compositions comprising thermolysin formulated and / or engineered to resist detergent-induced inactivation.

A preparing method of proteolytic peptides extracted from debitterized peach kernels includes the following steps of firstly, conducting squeezing and deoiling; secondly, extracting proteins; thirdly,conducting enzymolysis on the proteins; fourthly, conducting ultrafiltration separation; fifthly, conducting plastein reaction; sixthly, conducting centrifugation; seventhly, mixing components; eighthly, conducting incculated fermentation; ninthly, conducting freeze-drying. Enzymolysis is conducted on the proteins extracted from the peach kernels by means of thermolysin to form low-bitter-taste enzymolysis liquid, high-bitter-taste components are separated out through the ultrafiltration method, the plastein reaction is conducted under the effect of specific protease, bitter-taste hydrophobicproteins deposit and then centrifugally removed, polypeptide components with all molecular weights are combined, aspergillus oryzae and aspergillus niger are introduced for fermentation, and the bitter taste is further eliminated through a fermentation method. The prepared debitterized peach kernel proteolytic peptides belong to amino acid nutrient supplements, are good in taste, free of bitter taste and high in quality, are easier to absorb by human bodies compared with proteins, and have good market prospects.

The present invention provides methods and compositions comprising at least one thermolysin-like neutral protease enzyme with improved storage stability and / or catalytic activity. In some embodiments, the thermolysin finds use in cleaning and other applications comprising detergent. In some particularly preferred embodiments, the present invention provides methods and compositions comprising thermolysin formulated and / or engineered to resist detergent-induced inactivation.

The invention provides an immobilization method for thermolysin. The immobilization method comprises the following steps of: (1) activating carriers: impregnating aminated mesocellular foam silicon carriers (MCFs-NH2) in 0.5 to 2.5 mM of benzoquinone solution, carrying out oscillating activation for 1 to 3 hours, centrifuging, taking precipitate out, washing the precipitate, and dispersing the washed precipitate in MES-NaOH solution again to obtain activated carrier liquid; and (2) immobilizing enzyme: adding the activated carrier liquid in to the thermolysin in an enzyme adding amount of 40 to 100mg thermolysin / g MCFs-NH2 to obtain mixed liquid, performing electromagnetic stirring reaction on the mixed liquid in ice bath for 18 to 20 hours or irradiating the mixed liquid for 2 to 4 minutes under the condition of 20 to 50 W of microwaves at the temperature of between 0 and 7 DEG C, centrifuging, taking precipitate out, and washing the precipitate by the MES-NaOH solution and thus obtaining the immobilized thermolysin. The performance of immobilized thermolysin prepared by the method is greatly improved compared with that of free enzymes, and the immobilization time is greatly shortened from 20 hours to 3 minutes and is totally shortened by 399 times, so that the cost of the enzyme immobilization is lowered.

The invention discloses a method for refining protamine sulfate. According to the technical scheme adopted by the invention, the method comprises the following steps: performing enzymolysis on the protamine sulfate, performing quick protein liquid-phase separation to purify enzymatic hydrolysate, desalting a protamine high-salt solution to obtain high-purity protamine sulfate. A polypeptide mixture with average molecular weight of 1100Da, obtained by performing enzymolysis on the protamine with thermolysin has the capacity of neutralizing heparin; immune response caused after injection is remarkably lowered, and the toxic and side effects are relieved.

The invention belongs to the technical fields of analytical chemistry and electrochemical luminescence detection. The invention relates to an electrochemical luminescence sensor of cortex actin and a method for determining the cortex actin. A nanogold and graphene modified gold electrode is prepared by using electrochemical reduction, and peptide with an active group bipyridine ruthenium connection is self-assembled on the modified electrode, so that the electrochemical luminescence sensor is formed. When the cortex actin of a target object exists, the cortex actin is combined with the peptide to prevent thermolysin from cutting the peptide, and a strong electrochemical luminescence signal is generated; and otherwise, when no cortex actin exists, the thermolysin cuts the peptide, and the electrochemical luminescence signal is weakened. Therefore, the cortex actin is determined. The method has the characteristics of simplicity, high sensitivity, simple structure and low cost of an instrument device.

The present invention relates to the use of Thermolysin-Like Metalloproteases in cleaning processes, such as laundry and dish wash, and in particular to the use in low temperature wash and in removal of egg stains. The invention also relates to detergent compositions and cleaning compositions comprising Thermolysin-Like Metalloproteases.

Methods are disclosed for the reduction or elimination of bacterial biofilms on biological and non-biological surfaces, as methods for the treatment of wounds, skin lesions, mucous membrane lesions, and other biological surfaces infected or contaminated with bacterial biofilms using compositions comprising a synergistic combination of thermolysin and at least one aminoglycoside antibacterial agent.

An artificial gene method for preparing thermolysin includes configuring expression carrier-transformer colibacillus BL21(DE3), and inducing the expression of thermolysin. Its advantages are high output rate and activity of thermolysin.

The invention belongs to the technical field of biomedicine, and relates to a preparation method of injectable hydrogel, and an application of the injectable hydrogel to of postoperative tumor treatment. Thermolysin is used for catalyzing N-( 9-N-( 9-fluorenylmethyloxycarbonyl)-L-phenylalanine(Fmoc-F) and L- phenylalanine-L- phenylalanine-L-L-dopa(FF-Dopa) to form an Fmoc-F-FF-Dopa unit. The unitfurther utilizes hydrogen bonds, hydrophobic interaction, phi-phi accumulation and other non-covalent bond force, so that self assembly under the temperature condition being similar to the temperatureof a human body within a short time is performed to form the hydrogel. The enzyme catalysis cross-linking hydrogel has syringeability, and can be implanted into a cavity formed by a tumor operation as a medicine storing house, so that the carried medicines are slowly released in the local part of the cavity formed after the operation at a stable and controllable rate and with an appropriate concentration, drug effects are sufficiently exerted, remaining tumor cells are killed, and toxic and side effect caused by systemic administration can be avoided.

Methods for generating F(ab′)2 fragments from antibodies using thermolysin as well as F(ab′)2 fragments and compositions comprising F(ab′)2 fragments generated by the method are described.

The invention provides a method for amplifying natural killer cells from placenta tissues, which comprises the following steps: (1) taking perinatal placenta tissues, and cutting down villus lobule structure tissues in a small block form; (2) digesting placental lobule tissues by utilizing I and II type collagenase, DNA enzyme and thermolysin; (3) extracting mononuclear cells in the digested cell suspension and mononuclear cells in placenta blood by using a full-automatic separation device method; (4) inducing amplification of the NK cells in the mononuclear cells by utilizing cell factors such as IL-15, FLT3-L and OK432; and (5) carrying out biological identification on the amplified NK cells. The method disclosed by the invention has excellent technical effects as described in the specification.

It is intended to provide ACE inhibitory tripeptides which are not easily digested by digestive enzymes after being orally taken and thus have fewer tendencies to lose their ACE inhibitory activity in vivo. More specifically, 3 tripeptides having an ACE inhibitory activity and showing a hypotensive effect in an animal experiment are discovered from a thermolysin digestion product of sesame. These tripeptides respectively have amino acid sequences Leu-Ser-Ala, Val-Ile-Tyr and Leu-Val-Tyr and show an angiotensin converting enzyme inhibitory activity.

Methods of processing protein, methods of generating products with the processed protein, and products comprising and / or made with the processed protein. The methods of processing protein include hydrolyzing the protein with a proteolytic agent such as thermolysin to generate hydrolyzed peptides and, optionally, crosslinking the hydrolyzed peptides with a transglutaminase to generate crosslinked peptides. The methods reduce the allergenicity of allergenicity proteins such as β-lactoglobulin and casein. The methods of generating products with the processed protein include methods of making foams, emulsions, and / or food products with the processed protein. The products comprising and / or made with the processed protein accordingly include foams, emulsions, and food products. The foams, emulsions, and food products have decreased allergenicity compared to corresponding products made with non-processed proteins.

Methods are disclosed for the reduction or elimination of bacterial biofilms on biological and non-biological surfaces, as well as methods for the treatment of wounds, skin lesions, mucous membrane lesions, and other biological surfaces infected or contaminated with bacterial biofilms using compositions comprising thermolysin.

The present invention deals with the proteolytic enzyme thermolysin which tends to be unstable in aqueous solution. The invention provides methods and compositions to enhance the stability of dissolved thermolysin in aqueous solution. Thermolysin, crude thermolysin or a lyophilisate containing thermolysin and one or more salts, is contacted with an aqueous buffer with a low salt concentration and a first solution is formed. Subsequently, a further salt in solid form is added and dissociated, thereby forming a second solution comprising thermolysin in a stabilized form.

Methods are disclosed for the reduction or elimination of bacterial biofilms on biological and non-biological surfaces, as well as methods for the treatment of wounds, skin lesions, mucous membrane lesions, and other biological surfaces infected or contaminated with bacterial biofilms using compositions comprising thermolysin.

The present invention relates to the use of Thermolysin-Like Metalloproteases in cleaning processes, such as laundry and dish wash, and in particular to the use in low temperature wash and in removal of egg stains. The invention also relates to detergent compositions and cleaning compositions comprising Thermolysin-Like Metalloproteases.

The invention discloses a preparation method of an antioxidant peptide mixture. The preparation method comprises the following steps: by utilizing shrimp byproducts, carrying out two-step enzymolysis of alpha-chymotrypsin and thermolysin, and carrying out Sephadex G-15 gel filtration and DEAE Sephacel ion exchange chromatography separation to obtain an antioxidant peptide mixture with 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity of 10.54 + / - 0.03 mumol AAE / g pro and ferric ion reducing ability (FRAP method) of 89.29 + / - 0 .01 mumol AAE / g pro. The invention also discloses an antioxidant peptide mixture obtained by the method. The size of molecule is 500 to 1000Da. The preparation method and the antioxidant peptide mixture provided by the invention provide shrimp byproducts a new use way, the waste of resources and the pollution are reduced, and the commercial value and utilization rate of shrimp byproducts are enhanced.

Disclosed is a hydrogel comprising a hydrophilic gelling agent that includes a nonionic cellulose ether, and active thermolysin, wherein the proteolytic activity of the thermolysin does not decrease by more than 20% when stored at room temperature for 6 months.

An object of the present invention is to provide a method for producing a mesenchymal stromal cell composition, comprising conveniently and aseptically separating high-purity amnion-derived MSCs by performing enzyme treatment only once. According to the present invention, the following are provided: a method for producing a mesenchymal stromal cell composition, comprising: performing enzyme treatment of an amnion with collagenase and thermolysin and / or dispase; and filtering the enzyme-treated amnion through a mesh; a method for producing a cryopreserved mesenchymal stromal cell composition; and a therapeutic agent comprising as an active ingredient the mesenchymal stromal cell composition for a disease selected from graft-versus-host disease, inflammatory bowel disease, systemic lupus erythematosus, liver cirrhosis, or radiation enteritis.

The invention provides an immobilization method for thermolysin. The immobilization method comprises the following steps of: (1) activating carriers: impregnating aminated mesocellular foam silicon carriers (MCFs-NH2) in 0.5 to 2.5 mM of benzoquinone solution, carrying out oscillating activation for 1 to 3 hours, centrifuging, taking precipitate out, washing the precipitate, and dispersing the washed precipitate in MES-NaOH solution again to obtain activated carrier liquid; and (2) immobilizing enzyme: adding the activated carrier liquid in to the thermolysin in an enzyme adding amount of 40 to 100mg thermolysin / g MCFs-NH2 to obtain mixed liquid, performing electromagnetic stirring reaction on the mixed liquid in ice bath for 18 to 20 hours or irradiating the mixed liquid for 2 to 4 minutes under the condition of 20 to 50 W of microwaves at the temperature of between 0 and 7 DEG C, centrifuging, taking precipitate out, and washing the precipitate by the MES-NaOH solution and thus obtaining the immobilized thermolysin. The performance of immobilized thermolysin prepared by the method is greatly improved compared with that of free enzymes, and the immobilization time is greatly shortened from 20 hours to 3 minutes and is totally shortened by 399 times, so that the cost of the enzyme immobilization is lowered.

The present invention deals with the proteolytic enzyme thermolysin which tends to be unstable in aqueous solution. The invention provides methods and compositions to enhance the stability of dissolved thermolysin in aqueous solution. Thermolysin, crude thermolysin or a lyophilisate containing thermolysin and one or more salts, is contacted with an aqueous buffer with a low salt concentration and a first solution is formed. Subsequently, a further salt in solid form is added and dissociated, thereby forming a second solution comprising thermolysin in a stabilized form.