15 results about "Endoenzyme" patented technology

According to the invention, lactobacillus rhamnosus UP180 with relatively strong cellulase activity and lactobacillus paracasei ID001 with relatively strong protease activity are combined to prepare composite probiotic powder, the composite probiotic powder is used for jointly fermenting the endothelium corneum gigeriae galli formula, and meanwhile, a polypeptide capable of improving the digestive enzyme activity of Caco-2 cells is found from a fermentation product. The compound strain fermentation effectively decomposes macromolecular proteins in the endothelium corneum gigeriae galli formula and converts the macromolecular proteins into small molecular peptides, effective active ingredients in the formula are fully released, and the conversion effect is obviously superior to that of single strain fermentation. Besides, the endothelium corneum gigeriae galli composition leavening prepared by the invention can improve nutrient absorption of the rats by improving enzyme activity and metal ion chelation in intestinal tracts of the rats in the weaning period and promote growth of the rats in the weaning period, and has a potential application prospect in the field of nutrient absorption of children.

The invention relates to superoxide dismutase (SOD) as well as a preparation method and application thereof. By performing codon optimization on the SOD gene of the extreme microorganism geobacillus thermoalkane, a recombinant expression system is successfully constructed, high-efficiency expression of soluble protein is realized, and the concentration of protein in fermentation liquor reaches 200mg / L. The enzyme shows excellent stability under extreme conditions, the retention rate of enzyme activity exceeds 50% within the range of lasting at 80 DEG C for 2 h and pH 4.0-12, and the free radical scavenging capacity of the enzyme is improved by 20 times compared with that of vitamin C with the same concentration. The enzyme has a wide application prospect in the field related to cosmetics, the stability exceeds 95% in the environment of 60 DEG C, and the stability is higher than 75% in the pH range of 5.0-11.

The invention provides cyclomaltodextrin preferred to hydrolyze gamma-cyclodextrin. The amino acid sequence of the cyclomaltodextrin is SEQ ID NO: 1. The cyclomaltodextrin enzyme has the strongest hydrolysis effect on gamma-cyclodextrin, can better hydrolyze pullulan and soluble starch, has the optimal action temperature of 30 DEG C and the optimal pH value of 7.0, has higher enzyme activity in the ranges of 20-30 DEG C and pH value of 6.0-7.0, has better cold adaptability, and can be used for preparing the beta-cyclodextrin. The method has a good application prospect in the aspects of removing a cyclodextrin embedding medium, especially a gamma-cyclodextrin embedding medium, and preparing maltooligosaccharide and panose.

The invention provides high-temperature mannase and application thereof. The amino acid sequence of the high-temperature mannase is SEQ ID NO: 1. The mannase has the strongest hydrolysis effect on konjac gum, can hydrolyze guar gum and has a wide acting temperature and pH range, the enzyme activity is kept at 95% or above within the range of 45-75 DEG C, the enzyme activity is kept at 65% or above within the range of pH 5.5-8.5, and various common metal ions can promote the enzyme activity or have little influence on the enzyme activity. The enzyme is beta-mannase with endo-activity, and has a good application prospect in the aspects of feed processing, mannan oligosaccharide preparation, biological polishing, agricultural waste utilization, plant active substance extraction and the like under medium and high temperature conditions.

The invention relates to a method for preparing scDNA, which comprises the following steps: providing a circular nucleic acid template which is double-stranded and comprises a to-be-recombined ring-forming region and a to-be-enzyme-digested incision site arranged outside the region, and the to-be-recombined ring-forming region comprises an expression framework and a recombination site pair arranged on two wings of the expression framework; the present invention relates to a method for preparing a cyclic nucleic acid template, the cyclic nucleic acid template being amplified by RCA rolling circle amplification to form a concatemer, the concatemer comprising a tandem repeat unit of a cyclic nucleic acid template sequence, the concatemer being linearized by enzyme digestion in the incision site of the tandem repeat unit with a restriction enzyme simultaneously with or subsequent to the rolling circle amplification, the linearized concatemer comprising a cyclic nucleic acid template sequence, the cyclic nucleic acid template comprising a cyclic nucleic acid template sequence, and the cyclic nucleic acid template sequence comprising a cyclic nucleic acid template sequence. Free linearized nucleic acid is obtained; in a cell-free system, the linearized nucleic acid is recombined with a recombinase into scDNA comprising the expression framework. The method provided by the invention really realizes efficient preparation of scDNA.

The invention provides application of tetrathiomolybdate in preparation of drugs for inhibiting inflammatory factor storm, and belongs to the technical field of drugs for treating inflammatory factor storm and respiratory system diseases. The general molecular formula of the tetrathiomolybdate is X < + > MoS < 42-Y < + >, and the positions of ionic bonds of four sulfur ions of the tetrathiomolybdate are interchangeable; x + and Y + are the same or different cations; the positive ions are inorganic positive ions and / or organic positive ions which can be dissociated in a solution to generate MoS < 2-> or can be subjected to enzymolysis in a biological system to generate MoS < 2->. The invention discloses unique advantages of a single drug scheme in inflammatory factor storm and respiratory system related diseases, especially bacterial acute respiratory system inflammation, for the first time, and tetrathiomolybdate is used as a single active component for preparing the drug for treating bacterial pneumonia and non-viral acute lung injury / acute respiratory distress syndrome.

The invention discloses laccase and application thereof, and belongs to the field of bioengineering. The high-activity laccase Lac-11 is obtained through database mining, and compared with existing high-activity laccase BstCotA, the high-activity laccase Lac-11 has the following beneficial effects that 1, the enzyme activity is remarkably improved, the enzyme activity is improved by nearly 50% under the conditions that the temperature is 37 DEG C and the pH is 5.0, and the industrial enzyme cost is reduced; 2, the pH stability is higher, the enzyme activity retention rate is higher in the pH range of 5.0-8.0, and the method is suitable for wider acid-base environments. And 3, the thermal stability is better, the stability at the medium-high temperature interval of 50-80 DEG C is obviously higher than that of a control enzyme, and the method is suitable for an industrial high-temperature process. 4, the dye decolorization efficiency is higher, the decolorization rates of amino black 10B and crystal violet respectively reach 31% and 37%, and printing and dyeing wastewater can be efficiently treated. And 5, the lignin degradation capacity is outstanding, the degradation rates within 1 hour, 2 hours and 24 hours are 22%, 25% and 58% respectively, and pulping bleaching and biomass recycling are facilitated.

The application belongs to the technical field of mass spectrometry, and particularly relates to a mass spectrometry detection method and application of intracellular blood coagulation factor X, which comprises the following steps: specifically enriching ectopic synthesis of blood coagulation factor X in tumor cells to obtain a protein sample; performing protein sample fixation and treatment on the protein sample by using SDS-PAGE gel electrophoresis, placing the gel in an ice box after scanning the gel by using a gel imaging system, and storing the gel; performing in-gel enzymatic digestion on the blood coagulation factor X in the gel to obtain a peptide segment solution; detecting the endogenous synthesis of blood coagulation factor X in tumor cells by using mass spectrometry technology, and obtaining mass spectrometry data. The application firstly adopts the mass spectrometry technology to detect the blood coagulation factor X synthesized by tumor cells outside normal liver cells, specifically enriches the ectopic synthesis of blood coagulation factor X in glioma cells by using the protein immunoprecipitation technology, performs SDS-PAGE gel electrophoresis, performs in-gel enzymatic digestion, and detects the endogenous synthesis of blood coagulation factor X in tumor cells by using the mass spectrometry technology.

A cellulose-degrading bacterium (Trichoderma erinaceum) A18 and a culture medium and a preparation method of the culture medium, the bacterium is preserved in Guangdong Microbial Culture Collection Center, and the preservation number is GDMCC No. 65616, the bacterium is screened from soil and fallen leaves in Nanning City. The culture medium comprises a seed culture medium and an enzyme production culture medium, components of the seed culture medium are as follows: sodium carboxymethyl cellulose 10 g / L, protein peptone 3 g / L, potassium dihydrogen phosphate 4 g / L, magnesium sulfate heptahydrate 0.3 g / L and calcium chloride 0-0.3 g / L; components of the enzyme production culture medium are as follows: sodium carboxymethyl cellulose 8-12 g / L, protein peptone 2-3 g / L, yeast powder 0.2-3 g / L, ammonium sulfate 0.2-2 g / L, potassium dihydrogen phosphate 4 g / L, magnesium sulfate heptahydrate 0.3 g / L and zinc chloride 0.002 g / L. After the seed culture medium and the enzyme production culture medium are configured with distilled water, the seed culture medium and the enzyme production culture medium are placed in a high-pressure sterilization pot and sterilized at 121 DEG C, inoculation is carried out according to a proportion, and anaerobic culture is carried out in a closed state. The cellulose-degrading bacterium A18 can effectively improve cellulose endoenzyme activity by using the application.

The present application provides a high-temperature mannanase and its application, and the amino acid sequence thereof is SEQ ID NO:1. The mannanase of the present application has the strongest hydrolysis effect on konjac gum, and can also hydrolyze guar gum, has a wide temperature and pH range, the enzyme activity is maintained at 95% or above in the range of 45-75 DEG C, the enzyme activity is maintained at 65% or above in the range of pH 5.5-8.5, and various common metal ions can promote the enzyme activity or have little effect on the activity. The enzyme is a beta-mannanase with endo-activity, and has a good application prospect in feed processing, preparation of manno-oligosaccharides, biological polishing, utilization of agricultural waste, extraction of plant active substances and the like under medium-high temperature conditions.

The invention discloses a signal amplification system for detecting mitochondrial enzyme activity and application. The signal amplification system comprises a depurination and depyrimidine endonuclease 1 response module, an rRNA activation module and a signal report and amplification module. The depurination and depyrimidine endonuclease 1 response module contains a DNA duplex AP-I of an AP site, wherein the DNA duplex AP-I can be specifically recognized and cut by the depurination and depyrimidine endonuclease 1; the rRNA activation module contains a hairpin probe R-H1 of a mitochondrial rRNA recognition sequence; and the signal reporting and amplifying module comprises a fuel hairpin H2 marked with a fluorescence reporting group and a quenching group. The system provided by the invention successfully realizes real-time and in-situ imaging of APE1 enzyme activity in living cell mitochondria, and provides a powerful tool for researching related life processes at a subcellular level.

The invention relates to the technical field of enzymolysis, and particularly discloses a donkey-hide gelatin small molecule peptide production device and a production process, the donkey-hide gelatin small molecule peptide production device comprises a tank body, a rotating shaft, a stirring assembly and a ventilation assembly, an end cover is arranged at the top end of the tank body, the rotating shaft is arranged in the tank body, the stirring assembly is arranged on the rotating shaft, and the ventilation assembly is arranged in the upper area of the tank body; the ventilation assembly comprises a ventilation disc, movable plates, a sealing rod and a rotating seat, the ventilation disc is in sliding fit with the rotating shaft and is connected with the end cover through a lifting mechanism, a plurality of ventilation holes distributed at intervals are formed in the ventilation disc, the multiple movable plates are distributed in the circumferential direction of the ventilation disc at intervals, and each movable plate is connected with the ventilation disc through a first elastic piece; a plurality of sealing rods distributed at intervals are arranged on each movable plate, each sealing rod corresponds to one vent hole, the rotating seat is arranged on the rotating shaft, and a groove is formed in the rotating seat; the whole liquid level can be prevented from crusting, and crusts are broken and enter the liquid for enzymolysis.

The invention belongs to the technical field of daily chemicals, and particularly relates to a tableware detergent and a preparation method thereof. According to the detergent, the modified plant saponin derivative and the immobilized multienzyme complex liquid are adopted as main active components, the components such as the compound cleaning aid, the thickening agent and the hand care agent are supplemented, and the detergent has excellent dirt-removing power, enzyme stability and skin mildness. The modified plant saponin is grafted with a hydrophilic chain segment and an amphoteric auxiliary agent is introduced, so that the water solubility, emulsibility and system compatibility of the modified plant saponin are remarkably improved; the three enzymes are subjected to immobilization treatment to form a stable suspension, the adaptability is high, and the enzyme activity retention rate in the storage period is high. Performance tests show that the detergent has a good cleaning effect on grease, starch and protein composite dirt, has no irritation after being used, and is particularly suitable for families and infant tableware cleaning scenes.

The invention provides an application of OGA as a diagnostic target in GBC diagnosis and GBC prognosis products, relates to the technical field of biomedicine, and explores molecular changes, caused by PNI, of ecological nichs around nerves through combination of laser microdissection and a high-sensitivity mass spectrometry technology. As a result, an endoenzyme OGA involved in the glycosylation of O-GlcNAc is abnormally highly expressed in neurons after tumor invasion. In a non-contact co-culture system, the application finds that OGA can be delivered from tumor cells to neurons through EVs. Based on this, it is found that GBC cells transfer OGA to neurons through EVs, so that glycosylation of RIPK1 is inhibited, phosphorylation and activation of RIPK1 are promoted, and finally neuronal necroptosis is induced. RIPK1 is a core factor for regulating and controlling cell survival, inflammatory response and programmed death, and is activated through a compound IIb pathway under the action of exogenous OGA. The application also finds that the Ser440 site of the RIPK1 is a key region of OGA interaction, and the mutation of the Ser440 site can break the balance between glycosylation and phosphorylation so as to influence RIPK1 signal transduction.

The present application belongs to the technical field of genetic engineering and enzyme engineering, and particularly relates to a low-temperature neutral protease from marine microorganism colwellia psychrerythraea, a preparation method thereof and application of the low-temperature neutral protease in hydrolysis of proteins. In the present application, a protease gene from colwellia psychrerythraea is connected with an expression vector pPIC9K to obtain a recombinant plasmid, then the recombinant plasmid is electrotransformed into Pichia pastoris cells, and a target gene is expressed by using methanol induction, and a centrifugal product is obtained after removing supernatant, thereby obtaining a recombinant low-temperature neutral protease. The recombinant low-temperature neutral protease can be used for hydrolysis of fish meal, and the recombinant low-temperature neutral protease has higher enzymatic activity in the range of 15 DEG C to 35 DEG C, and still maintains more than 60% at 15 DEG C, so that the recombinant low-temperature neutral protease is a neutral low-temperature metal protease which can maintain higher activity at low temperature or normal temperature.