33results about How to "Retain enzyme activity" patented technology

The invention relates to a magnetic composite microsphere immobilized laccase and a preparation method thereof, and belongs to the field of environmental microbiology. In the immobilized laccase, metal-chelated magnetic composite microspheres are taken as a carrier, and laccase is immobilized on the carrier through coordination bonding action. The preparation method comprises two steps of: preparing the carrier, namely synthesizing magnetic ferroferric oxide nanoparticles by using a chemical precipitation process, preparing core-shell magnetic silica by taking the nanoparticles as cores through a sol-gel method, grafting polymer containing carbonyl or amino on the surface of the magnetic silica by taking gamma-chloropropyltrimethoxysilane as a medium, and preparing a functional carrier capable of immobilizing the laccase by chelating transition metal ions (such as Cu2+. Zn2+, Ni2+ or Co2+); and immobilizing the laccase, namely performing coordination adsorption of the laccase on the surface of the carrier, washing, and drying to obtain the finished product of immobilized laccase. The immobilized laccase prepared by the method has high stability and activity and quick magnetic response, is easily recovered from a reaction system and can be repeatedly used; and the preparation process is simple and the cost is low.

The present invention discloses a mixed enzyme production process by solid fermentation of fruits and vegetables and microorganisms and the production process includes the following steps: step one, washing and cleaning fresh fruits and vegetables; step two, juicing the cleaned fruits and vegetables using a juice extractor and obtaining the fruit and vegetable juice and the fruit and vegetable residue; step three, adding water content adjusting raw materials according to different water content of the obtained fruit and vegetable residue to adjust the water content of the fruit and vegetable residue, and mixing the above materials evenly to obtain a base material; step four, putting the step three obtained base material into a fermentation bottle to conduct sterilization; step five, cooling the sterilized fruit and vegetable base material to room temperature, and inoculating microorganism liquid strains to conduct solid fermentation; and step six, after the end of solid fermentation, blending the obtained fruit and vegetable juice into the solid fermented materials, conducting low temperature drying, and thereby obtaining the fruit and vegetable and microorganism mixed enzymes containing both fruit and vegetable enzymes and microbial enzymes. The mixed enzymes can not only fully preserve the enzyme activity of fruits and vegetables, but also obtain the enzyme ingredients of microorganisms, and in addition, the production process can also achieve the full advantage of the entire fruits and vegetables, does not produce any waste, and is ecological and environmental protective.

The invention relates to the technical field of the food deep processing and particularly relates to black garlic as well as a fermentation process thereof. The fermentation process of the black garlic comprises the following steps of freezing fresh garlic sections at a freezing temperature being -10 DEG C to -40 DEG C for less than 48 hours; and fermenting through two stages, wherein constant-temperature fermentation is carried out for 2-10 days at the first-stage fermentation temperature being 10 DEG C-50 DEG C, and the constant-temperature fermentation is carried out for 4-12 days at the second-stage fermentation temperature being 70 DEG C-90 DEG C. According to the fermentation process disclosed by the invention, the content of substances such as reducing sugar, total phenols and amino acid in the garlic can be greatly increased without adding any additive, and the black garlic fermentation period is shortened, so that the whole fermentation process can be completed in only 8-14 days; the obtained black garlic has characteristics of being good in flavor, elastic in texture, high in nutritional ingredient, high in safety, easy to store, and the like.

The invention provides a method for preparing a health-care enzyme drink from grapefruits and belongs to the technical field of bioengineering technologies. According to the method, the grapefruits are taken as a main fermentation raw material, are peeled and mashed to be pulpy, raw honey and purified water are added, nectariferous bacteria, lactobacillus casei and lactobacillus plantarum strains are taken as fermentation strains, a fermentation liquid which is 1%-5% of the volume of a fermentation stock solution is added to a fermentation system, the fermentation temperature is 25-35 DEG C, pH ranges from 5.0 to 8.0, the fermentation time lasts for about 10d, and the health-care grapefruit enzyme drink is prepared. According to the method, complete fermentation is realized, various enzymes, vitamins, amino acid and micro elements are sufficiently released, the product has a unique flavor of grapefruits, is palatably sweet and sour and has rich nutrition, and a traditional fermentation technique is adopted.

The embodiment of the invention discloses a probiotic enzymatic composition capable of regulating intestinal tract. The composition is prepared by mixing the following raw materials in percentage by weight: 65 to 75 percent of dietary fiber, 10 to 15 percent of comprehensive vegetable and fruit enzymatic powder, 10 to 15 percent of soybean protein powder, 2.5 to 4.5 percent of probiotic bacteria,1 to 2 percent of plant protease, 0.3 to 0.8 percent of soybean phospholipid, 0.5 to 0.8 percent of an acidifier and 0.08 to 0.15 percent of a sweetener. The probiotic enzymatic composition provided by the invention has a remarkable curative effect in regulating intestinal problems, can effectively prevent dyspepsia and improve constipation, and has the functions of protecting intestinal mucosa and enhancing immunity, thereby fundamentally solving intestinal problems. The composition provided by the invention is natural, free from side effects, and safe and effective in use, and has a broad market application prospect.

The invention discloses an arginine deiminase mutant from an arthritis-type mycoplasma, a gene, a polyethylene glycol modified substance and an application thereof. The arginine deiminase mutant is a dipolymer which is formed by two protein subunits, wherein amino sequences of the protein subunits are represented as SEQ ID No.2 in a sequence table. The arginine deiminase mutant is increased in expression quantity in escherichia coli, is increased in renaturation efficiency, is not changed in enzyme activity and is suitable for industrial production. The polyethylene glycol modified substance of the arginine deiminase mutant can be used for preparing anti-tumor drugs, such as anti-hepatoma drugs, anti-leukemia drugs and anti-melanoma drugs.

The invention provides a method for preparing an enzyme beverage from polygonum perfoliatum leaves. The method includes the steps that the polygonum perfoliatum leaves are used as a main fermentation raw material, raw honey and natural springwater provided by the Wuxi hundred-year Shangshui drinks limited company are added, nectar source bacterium, lactobacillus casei and lactobacillus plantarum strains are adopted as fermenting strains, fermenting bacterium liquid is added into a fermentation system, wherein the volume of the fermenting bacterium liquid accounts for 1-5% of the volume of a fermenting stock solution, and fermentation is carried out for about 10 d at the temperature of 25 DEG C-35 DEG C and the pH value of 5.0-8.0 to prepare the polygonum perfoliatum leaf enzyme healthcare beverage. Fermentation of the beverage is complete, various enzymes, vitamins, amino acid and microelement are fully released, and the beverage has the unique flavor of the polygonum perfoliatum leaves, is rich in nutrition and tasty and belongs to the technical field of bioengineering by the adoption of a traditional fermentation technology.

The invention discloses a method for preparing a super paramagnetic carrier of a cross-linking immobilized lipase. The preparation of the super paramagnetic carrier is realized by preparing a nano-magnetic microsphere Fe3O4, coating the nano-magnetic microsphere Fe3O4, functionally modifying the coated nano-magnetic microsphere Fe3O4. A thermo-sensitive material, namely N-isopropylacrylamide, is introduced into the super paramagnetic carrier prepared according to the method, and the structural performance of the carrier can be changed through temperature control, so that the lipase which is immobilized through the super paramagnetic carrier can get out of the interface performance control to realize direct reactions among alcohols, acids and lipids, and separation of the reaction products is facilitated; moreover, lipase activity control through alpha coverage is facilitated. The super paramagnetic carrier is a super paramagnetic polymer microsphere based on the mode of lipase interface activity action; the lipase can be immobilized through the super paramagnetic polymer microsphere; the immobilized lipase is high in magnetic separation performance; 90% or higher of the lipase activity is kept; the detect that the lipase activity in an enzymatic reaction depends on the interface effect is overcome.

The invention provides a maca concentrate, abelmoschus manihot and active rhizoma polygonati combined food and a preparation method thereof. The preparation method comprises the following steps: 1, smashing maca concentrate until the granularity is 300 meshes; 2, smashing active abelmoschus manihot until the granularity is 300 meshes; 3, smashing active rhizoma polygonati until the granularity is 300 meshes; 4, mixing and pelletizing maca concentrate powder which accounts for 60-70% of the total weight, the active abelmoschus manihot powder which accounts for 15-20% of the total weight and the active rhizoma polygonati powder which accounts for 15-20% of the total weight at a set normal temperature of 15-30 DEG C, and tabletting at a set pressure of 10kg/g-15kg/g, thereby obtaining the maca concentrate, abelmoschus manihot and active rhizoma polygonati combined food. According to the preparation method, the active abelmoschus manihot and the active rhizoma polygonati which are obtained by an ultralow-temperature freeze drying method are adopted, all active ingredients of the abelmoschus manihot and the rhizoma polygonati are preserved, especially active enzymes and relevant enzymein the abelmoschus manihot and the rhizoma polygonati are preserved. The preparation method is simple, low in cost and convenient to operate.

The invention provides a method for making geranium skin care enzymes with nectar-derived bacteria. The technological feature is to use geranium as the main raw material, and the preservation number preserved in the China Typical Culture Collection Center on June 24, 2013 is: CCTCC NO: M2013283 Bacillus amyloliquefaciens H47 was the fermentation strain of Bacillus amyloliquefaciens. According to the volume ratio of geranium: raw honey: pure water is 2‑5: 1‑3: 7‑2, add it and mix it evenly to make a geranium fermentation stock solution, add the nectar source bacteria seed liquid to the fermentation stock solution according to the ratio of 1‑5%. After fermenting for 6-14 days at a temperature of 25°C-35°C and a pH value of 5.0-8.0, the crude product of geranium skin care enzyme can be obtained. Preliminarily screen out large particle residues through a coarse filter with a number of 20-32 meshes, and then further filter out small particle residues through a fine filter with a number of 80-100 meshes to obtain the geranium skin care enzyme.

The invention discloses a preparation method and application of nanogel with a hunger combined gas therapy anti-tumor function. The nanogel is prepared by the following steps: firstly, performing electrostatic interaction on polyethyleneimine azide grafted with L-arginine and glucose oxidase to form a nano-enzyme compound; and then covering the surface of the nano-enzyme compound with an alkynyl-containing polycarbonate block copolymer, and performing click chemical crosslinking on azide and alkynyl. Compared with the prior art, according to the method disclosed by the invention, specific enzymatic reaction is utilized to consume endogenous glucose and release nitric oxide gas, cancer cells are killed through hunger and gas therapy, besides, the nitric oxide gas can redistribute oxygen in the cancer cells, hypoxia of tumors is relieved, and wide application prospects are realized in the aspects of resisting tumors and inhibiting tumor metastasis.

The invention provides a preparation method of graphene immobilized laccase. The graphene immobilized laccase is prepared by immobilizing laccase to a carrier of two-dimensional crystal material having excellent conductivity through nonspecific adsorbing action. The preparation method comprises the steps of preparing the carrier and immobilizing laccase; particularly, the carrier, graphene, is prepared by using a phenol as a raw material to carry out graphitization on sandwiched FePO4/dodecylamine nanolayer composite, and stripping before synthesis; the immobilized laccase is immobilized by adsorbing laccase to the surface of the evenly dispersed graphene in nonspecific manner, centrifuging, washing, and drying to obtain the finished immobilized laccase. The carrier of graphene herein helps increase electron transfer rate of enzymatic catalytic reaction process and improve reaction efficiency; the immobilized laccase prepared via the preparation method has high fixing quantity, high activity and good stability, can be recycled from the reaction system, and is reusable; the preparation process is simple and feasible.

The invention provides a production method of navel orange compound enzyme tea. The navel orange compound enzyme tea is prepared as follows: navel oranges, pineapples and apples rich in enzyme components are taken as main materials, white granulated sugar and white vinegar are added, alcohol fermentation is firstly performed with yeast, lactic acid bacteria are added for enzyme fermentation, and treatment is performed. Complete fermentation is realized, so that multiple enzymes, vitamins, amino acids, microelements and enzyme components originally contained in selected composition materials ofthe product are fully released, small molecular substances easy to absorb by the human are obtained, excellent strains and advanced process technologies are selected in production of the product, conversion and synthesis of enzyme can be effectively promoted in the fermentation process, so that the product has significant effects of regulating gastrointestinal functions, relaxing bowels, helpingdigestion, removing toxins, maintaining beauty and the like and is convenient to use and carry. The tea is produced with the traditional fermentation technology and belongs to the technical field ofbioengineering.

The invention relates to a processing method of aged health-care old white tea. The processing method comprises the steps of (1) performing picking; (2) performing withering; (3) performing piling; (4) performing drying; (5) performing storing; (6) mixing with dried orange peel powder; and (7) performing styling and drying. According to the method, the aged health-care old white tea is prepared bya white tea technology, the making method of the white tea is unique in technology, and the white tea does not need to be fried or kneaded, so that the permeability of cell membranes and enzymatic activity can be reserved to the maximum extent, leaf cells are subjected to a series of slow hydrolyzing and oxidation reaction under the situation that the leaf cells are not damaged or slightly damaged, and conjugated non-volatile precursor substances are converted into free volatile aroma components under the action of hydrolase, and foundation is established for unique color, fragrance and tastewhich are formed in the later stage.

The present application discloses novel PH20 variants or fragments thereof, human hyaluronidase having improved thermal stability and enzymatic activity, wherein the hyaluronidase is an enzyme that hydrolyzes hyaluronic acid, in particular to PH20 variants or fragments thereof comprising one or more amino acid residue substitutions in a variant having the amino acid sequence of SEQ ID NO: 3, and one or more amino acid residues are selectively deleted at the N-terminus and/or C-terminus.

The invention discloses a method for extracting a soil mircoorganism carbon-sequestrated enzyme, comprising the following steps: A: adding soil to a precooled ion remover and a phosphate buffer in sequence; evenly blending, carrying out centrifugation on the mixture at room temperature and collecting sediment; B: adding the sediment obtained in the step A to a cell sample protein extracting solution, carrying out vortex blending, breaking cells by using an ultrasonic method in ice-bath, carrying out centrifugation to collect supernate, adding solid ammonium sulphate until the solubility is reached, oscillating while blending, carrying out centrifugation to collect precipitated protein, adding a dissolved solution to dissolve the protein, and preserving the dissolved protein; and C: placing 1,5-ribulose diphosphate carboxylase/oxygenase protein solution obtained in the step B in water bath to recover enzymatic activity and using an ultraviolet spectrophotometer to measure the activity of 1,5-ribulose diphosphate carboxylase/ oxygenase and the nanomole enzymatic activity of fixed CO2 in each kilogram of the soil. The simple, convenient and fast method has the advantage of good repeatability of measurement results and suitability for extraction and activity measurement of soil mircoorganism carbon-sequestrated key enzyme 1,5-ribulose diphosphate carboxylase/oxygenase.

The invention discloses a preparation method of an exosome bionic preparation for synergistically promoting wound healing and a preparation thereof. The preparation method of the preparation comprises the following steps: preparing catalase-photosensitizer micelle, preparing ROS response lipidosome carrying catalase-photosensitizer micelle, extracting, separating and purifying exosome, mixing the lipidosome with the exosome, mediating membrane fusion between the liposome and the exosome by an extrusion method, and mixing the membrane fusion carrier and gel, and uniformly stirring the mixture to prepare the preparation. According to the preparation prepared by the invention, photodynamic therapy and biological therapy are combined, catalase-photosensitizer micelles are wrapped by a membrane fusion carrier, accumulated hydrogen peroxide is eliminated on a wound surface, oxygen is supplemented, and the photosensitizer generates controllable singlet oxygen to resist inflammation and sterilize, so that favorable conditions are created for treating the wound surface by the exosome; meanwhile, the gel provides a favorable moist environment for wound healing, and the gel is high in safety, good in stability, convenient to apply, simple in processing technology, low in cost and good in application prospect.

The invention relates to a method for producing continuously 4-hydroxycinnamyl alcohol using a multi-strain immobilized cell composition. Strain A used herein is recombinant strain E. coli M15-4Cl-CCR, and strain B is recombinant strain E. Coli M15-CAD; the method comprises the specific steps: 1, culturing strains; 2, immobilizing the strains to obtain respectively strain A immobilized cell particles and strain B immobilized cell particles; 3, producing 4-hydroxycinnamyl alcohol with the immobilized cells. The 4-hydroxycinnamyl alcohol is produced using modern bioengineering technology, and the application of the method for producing continuously with the multi-strain immobilized cell composition in the synthesis of plant-origin metabolite has important practical significance; high-performance liquid chromatography tandem mass spectrometry detection is established in the meantime, and more effective analytical means is provided for the aromatic polymers.