40results about How to "Improve the stability of enzyme activity" patented technology

The invention relates to genetic engineering field, in particular to eosinophilic lactase BGALA, gene and application thereof. The invention provides a lactase BGALA of eosinophilic Bispora sp.; the amino acid sequence of the lactase BGALA is shown in SEQ ID NO.1; and the invention provides genome and cDNA coding gene bgalA for coding the lactase. The invention obtains an eosinophilic lactase with the most proper pH value of 1.5, which maintains high enzymatic activity in acid environment, has better pH stability and heat stability, and antiprotease hydrolysis. The eosinophilic lactase can effectively hydrolyze the lactose in milk in simulated digestive tract environment. The eosinophilic lactase BGALA with the properties can be used for treating lactose intolerance, diary industry no processing cow milk and whey.

The invention discloses a preparation method of a beer liquid compound enzyme and belongs to the technical field of preparation of liquid compound enzymes. According to the preparation method, a liquid enzyme preparation for beer brewage is taken as a main raw material, stabilizers such as sugar, polyhydric alcohol, an amino acid, a thickener, albumin, metal ions and the like as well as substances such as a hop extract, a Chinese herbal medicine extracting solution, antibacterial peptides and the like which have anti-corrosion effects and can promote the enzyme preparation to be more stable are compounded scientifically, and the addition of a preservative is completely replaced, so that the breeding and the growth of harmful microorganisms are effectively inhibited, the activity of the liquid compound enzyme can be stabilized synergistically together with other components, the biological stability and the enzyme activity stability of the liquid compound enzyme are improved fundamentally, and the shelf life of the liquid enzyme preparation is effectively prolonged. The total number of bacterial colonies is in a range of 168-214 CFU / mL if the prepared liquid compound enzyme is stored at the room temperature for 12 months; enzyme activity loss ratios are in ranges of 2-2.5% and 1-1.7% respectively if the prepared liquid compound enzyme is stored at temperatures of 40 DEG C and 0 DEG C for 12 months.

The invention is suitable for the technical field of cleaning supplies and provides a washing machine slot cleanser and a preparation method and application thereof. The washing machine slot cleansercomprises the following components by weight: 2-5 parts of an anionic surfactant, 24-29.5 parts of a basic inorganic cleaning agent, 10-17 parts of a builder, 1.5-2.5 parts of an enzyme preparation and 0.5-1 part of an enzyme stabilizer, wherein the enzyme preparation is one or a mixture of at least two of protease, lipase, pectinase, cellulase, hemicellulase, mannase, staphylococcal nuclease, catalase and lysozyme. The enzyme preparation and the anionic surfactant has a coordinated function; under the action of the enzyme stabilizer, the enzyme activity stability of the enzyme preparation ishigh; various stains in a washing machine slot can be effectively decomposed and removed; the cleaning effect is good; cleanliness and no residues are achieved after cleaning; secondary pollution cannot be caused; and sanitation and hygiene in a washing machine is guaranteed.

The invention discloses a bromelain antiallergic food and a preparation method thereof. The bromelain antiallergic food disclosed by the invention is a bromelain beverage mainly prepared from bromelain, glucose, citric acid, VB1, L-cysteine and NaCl. By adding glucose into a formula of the bromelain antiallergic food, the enzyme activity stability of a bromelain solution is superior to the stabilities of bromelain solutions of other sugar solutions, so that the enzyme activity is beneficially guaranteed; the other raw materials in the formula do not contain sour substances, and citric acid is added, so that the taste can be well changed, and meanwhile, citric acid is capable of weakening the heat resistance of microorganisms, inhibiting the growth of the microorganisms and improving the performance of an antioxidant; and the bromelain has maximum inhibition ratio to hyaluronidase due to the concentration of the bromelain in the formula, and the enzyme activity decay is within an optimal enzyme activity equivalent range when the concentration is 0.2g / 100mL.

The invention provides a 1, 5-sorbitan determination kit, a preparation method and an application thereof. The kit contains a reagent R1 and a reagent R2, wherein the reagent R1 contains the followingcomponents: a buffer solution, adenosine disodium triphosphate, hexokinase, glucose-6-phosphate dehydrogenase, nicotinamide adenine dinucleotide, phosphoenolpyruvate monopotassium salt, 4-amino antipyrine (4-AA), pyruvate kinase, peroxidase, lactic dehydrogenase, ascorbic acid oxidase, magnesium chloride, potassium chloride, an enzyme stabilizer and a preservative; and the reagent R2 is preparedfrom the following components in concentration: the buffer solution, pyranose oxidase, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methylaniline sodium salt, a surfactant and the preservative. The kit is aliquid kit with strong stability, high accuracy, high sensitivity and a stable glucose elimination capability, can be suitable for a full-automatic biochemical analyzer, and is simple and convenient to operate.

The invention discloses a mutant of D-amino acid oxidase, the sequence of which comprises amino acid residue N at position 54 of the sequence shown in SEQ ID NO.1 or a sequence having at least 76% identity with SEQ ID NO.1, The mutated sequence of the 58th amino acid residue F, the 211th amino acid residue C and the 213th amino acid residue M; the D-amino acid oxidase mutant has a higher enzyme than the wild-type D-amino acid oxidase activity, enzymatic stability and / or ammonium resistance. The invention also discloses the application of the D-amino acid oxidase mutant in the preparation of 2-oxo-4-(hydroxymethylphosphinyl)butyric acid. The D-amino acid oxidase mutant of the present invention has high enzyme activity, improved enzyme activity stability and / or enhanced ammonium resistance, thereby reducing costs and being beneficial to industrial production.

The invention provides an enzyme manufacture method, which comprises the following working procedures of: complex acquisition: enabling a solid enzyme and a high-molecular adsorbing material to be in contact under the condition that the pH value is 6-8 to obtain a solid enzyme-high-molecular adsorbing material complex; contact: enabling the solid enzyme-high-molecular adsorbing material complex to be in contact with a reducing sugar; and separation: carrying out separation from a mixed system obtained in the working procedure of the contact to obtain the enzyme, wherein the high-molecular adsorbing material comprises at least one of the materials selected from the group consisting of a non-polar macroporous adsorbent resin, a non-polar skeleton high-molecular polymer and a non-ionic hydrophobic cross-linked polymer. The enzyme manufacture method can effectively prevent enzyme activity from being reduced, provides the enzyme with high enzyme activity and excellent enzyme activity stability, prolongs the service life of the enzyme, has simple manufacture working procedures, is easy to control the process, can meet the requirements of high-efficiency and stable production, and effectively reduces the production cost of enterprises.

The invention provides a preservation method of phospholipase C. By using the preservation method, the phospholipase C has good enzyme activity stability. When the preservation method of the phospholipase C provided by the invention is used in the practical application process of the phospholipase C, the production energy consumption can be reduced; and meanwhile, the activity loss of the phospholipase C is avoided.

The invention relates to a glutamate decarboxylase mutant with improved thermal stability and an application thereof, belonging to the field of bioengineering. The mutant is prepared primarily by the following steps: performing saturated mutation of glutamine Q, valine V and threonine T on the sites 5-7 of the amino acid sequence of glutamate decarboxylase, and screening to obtain high-stability mutants Q5E / V6S / T7V, Q5Y / V6R / T7K and Q5N / V6Y / T7V. In the glutamate decarboxylase mutant prepared in the invention, the half-inactivation temperature is 45-50.5 DEG C which is 4.9-10.2 DEG C higher than that of wild type mutant; and the half-life period at 45 DEG C is 76-129min, which is 3.2-4.3 times higher than 24min of wild type mutant. By transforming glutamic acid for 12h with the whole cell of glutamate decarboxylase mutant synthesized from recombinant escherichia coli, 260-350g / L gamma-aminobutyric acid (GABA) can be obtained, and the molar yield is 76.6-97.8%. In the glutamate decarboxylase mutant prepared in the invention, the thermal stability is obviously improved, the production of gamma-aminobutyric acid is facilitated, and a foundation is laid for efficient synthesis of gamma-aminobutyric acid.

The invention discloses a preparation method of immobilized beta-glucosidase in the field of enzyme preparations. According to the method, fructose is adopted as a pore-foaming agent, and the immobilized beta-glucosidase is prepared through the tetraethoxysilane hydrolysis reaction sol-gel method. Different immobilized beta-glucosidase samples are prepared by adding fructose water solutions with different concentrations and beta-glucosidase of different dosages. For the obtained immobilized beta-glucosidase, the invalid adsorption in the enzymolysis process of cellulose can be avoided, so that the enzymolysis conversion efficiency of cellulose is obviously improved, the immobilized enzyme has the advantages of good mechanical strength and multiple times of recycling, and the hydrolysis and saccharifying costs of cellulose are reduced.

The invention discloses a freeze-drying protective agent of glycosaminoglycan synthetase and application of the freeze-drying protective agent. The freeze-drying protective agent is prepared from trehalose, hydroxypropyl-beta-cyclodextrin and ectoine. The formula of the freeze-drying protective agent is different from the formula of the traditional freeze-drying protective agent, the components of the formula are simple, the freeze-dried glycosaminoglycan synthase is loose in structure, regular in shape and appearance and good in solubility, and particularly, the enzyme activity of the freeze-dried glycosaminoglycan synthase is slightly different from that of the freeze-dried glycosaminoglycan synthase before freeze-drying. And the enzyme activity can be kept above 90% after being stored at 4 DEG C and 25 DEG C for 90 days, which indicates that the freeze-drying protective agent can effectively reduce the activity loss of the glycosaminoglycan synthase in the freeze-drying process, and is beneficial to the storage and transportation of the glycosaminoglycan synthase.

The invention discloses a high-activity peanut shell polysaccharide and a preparation method thereof and belongs to the technical field of natural product preparation. Mutants of an endoglucanase and a cellobiohydrolase are obtained by a site-directed mutation way, compounding is performed, and an active polysaccharide and a flavonoid compound are synchronously obtained from peanut shells by an ultrasonic assisted enzymolysis and extraction way. The optimum hydrolysis temperature and enzyme activity of the complex enzymes are improved by the complex enzymes and the preparation process, operation steps are simplified, extraction time is reduced, synchronous preparation of the flavonoid compound and the water-soluble polysaccharide from peanut shells is realized, synchronous enzymolysis and extraction of the polysaccharide under a medium-high temperature condition is also realized, the obtained active polysaccharide is high in flavonoid content, and strong in antioxidant activity and antibacterial activity, and has a plurality of physiological functions and a wide application range.

The invention discloses a beer liquid compound enzyme and a preparation method thereof and belongs to the technical field of preparation of liquid compound enzymes. According to the beer liquid compound enzyme, a liquid enzyme preparation for beer brewage is taken as a main raw material, stabilizers such as sugar, polyhydric alcohol, an amino acid, a thickener, albumin, metal ions and the like as well as substances such as a hop extract, a Chinese herbal medicine extracting solution, antibacterial peptides and the like which have anti-corrosion effects and can promote the enzyme preparation to be more stable are compounded scientifically, and the addition of a preservative is completely replaced, so that the breeding and the growth of harmful microorganisms are effectively inhibited, the activity of the liquid compound enzyme can be stabilized synergistically together with other components, the biological stability and the enzyme activity stability of the liquid compound enzyme are improved fundamentally, and the shelf life of the liquid enzyme preparation is effectively prolonged. The total number of bacterial colonies is in a range of 168-214 CFU / mL if the prepared liquid compound enzyme is stored at the room temperature for 12 months; enzyme activity loss ratios are in ranges of 2-2.5% and 1-1.7% respectively if the prepared liquid compound enzyme is stored at temperatures of 40 DEG C and 0 DEG C for 12 months.

The invention provides a preservation method of phospholipase C. By using the preservation method, the phospholipase C has good enzyme activity stability. When the preservation method of the phospholipase C provided by the invention is used in the practical application process of the phospholipase C, the production energy consumption can be reduced; and meanwhile, the activity loss of the phospholipase C is avoided.

The invention relates to novel high-temperature resistant alpha-amylase, a preparing method of the novel high-temperature resistant alpha-amylase and an application of the novel high-temperature resistant alpha-amylase. As PCR site-specific mutagenesis is carried out on genes of wild-type high-temperature resistant alpha-amylase, and the genes are efficiently expressed in bacillus subtilis. The novel high-temperature resistant alpha-amylase is high in stability in the high-temperature environment and the acid environment, and can better adapt to industrial production to achieve the effects of saving energy, reducing consumption and improving efficiency. By means of the technical scheme, the genes of the wild-type high-temperature resistant alpha-amylase are separated from bacillus licheniformis, mutagenesis is carried out on His316 amino acid residues of the genes, efficient expression is carried out in the bacillus subtilis, and under the 90-DEG C condition and the pH-4.5 condition, the stability of the novel high-temperature resistant alpha-amylase (His316 to Arg) is improved compared with the wild-type high-temperature resistant alpha-amylase.

The invention relates to genetic engineering field, in particular to eosinophilic lactase BGALA, gene and application thereof. The invention provides a lactase BGALA of eosinophilic Bispora sp.; the amino acid sequence of the lactase BGALA is shown in SEQ ID NO.1; and the invention provides genome and cDNA coding gene bgalA for coding the lactase. The invention obtains an eosinophilic lactase with the most proper pH value of 1.5, which maintains high enzymatic activity in acid environment, has better pH stability and heat stability, and antiprotease hydrolysis. The eosinophilic lactase can effectively hydrolyze the lactose in milk in simulated digestive tract environment. The eosinophilic lactase BGALA with the properties can be used for treating lactose intolerance, diary industry no processing cow milk and whey.