75 results about "Phosphatidase" patented technology

The invention relates to a method for preparing phosphatidylserine abundant in polyunsaturated fatty acid and belongs to the technical field of bioengineering. The method is characterized by comprising the following steps of: firstly, catalyzing ester exchange reaction between phosphatide and polyunsaturated fatty acid ester by utilizing one or a mixture of phosphatidase A and lipase to generate phosphatide abundant in polyunsaturated fatty acid; and then catalyzing phosphor-transfer esterification reaction between the phosphatide abundant in the polyunsaturated fatty acid and L-serine by utilizing phosphatidase D to generate the phosphatidylserine abundant in the polyunsaturated fatty acid. The method has the advantages of no discharge of waste water, good product quality, no solvent residue, safe process operation, few reaction byproducts, no waste generation, cost reduction, simple production process and easy realization of scale production because of utilizing two enzymes to perform sub-step catalysis and perform reaction in the same reactor, and completing the reaction process in a non-solvent system. Therefore, the invention provides a good and feasible method for preparing the phosphatidylserine abundant in the polyunsaturated fatty acid.

The invention relates to a method for digesting grease through microwave pretreatment and an aqueous enzymatic method. The method includes soaking raw materials, performing microwave puffing treatment on camellia oleifera seed slurry, digesting grease with water, performing physical refining on the obtained grease after degumming through a phosphatidase enzymatic method, or directly performing rough filtration, refined filtration and nanofiltration, and then performing vacuum dehydration and drying to obtain grease products and a by-product of wet meal. According to the method, the microwave puffing treatment is adopted on raw materials such as camellia oleifera seeds, cell structures of the raw materials can be fully destroyed, and the oil yield of the following water immersion method can be improved; grease digestion through water after microwave pretreatment for materials belongs to an environment-friendly and safe production process; and the method has the advantages of being simple and environment-friendly in process, high in oil yield, low in investment and prone to industrial production.

The invention provides a method for refining rape seed crude oil by immobilized phosphatidase A1, and relates to the method for refining the rape seed crude oil. The problems of inactivation of enzyme caused by an uneasily controlled enzyme reaction condition during production and high cost of phosphatidase in enzyme process degumming can be solved by selecting the immobilized phosphatidase A1 for degumming. The method for refining the rape seed crude oil by the immobilized phosphatidase A1 is realized by the following steps: 1, preparing a reaction solution; 2, preparing a mixed solution; 3, preparing granulated immobilized enzyme; 4, pretreating the rape seed crude oil; 5, adding the immobilized enzyme into the oil; and 6, stirring and centrifugating to realize the refined rape seed crude oil by the immobilized phosphatidase A1. The rape seed crude oil is subjected to the degumming treatment by the method to improve degumming efficiency. Compared with the prior method, the method improves the degumming efficiency by 5 to 30 percent and the economic benefit by 1 to 3 percent.

The invention discloses a method for degumming vegetable fat by using phosphatidase A2. The method comprises the following steps of: A, heating crude oil to 40-50 DEG C; B, adding a 45 percent (W / W) citric acid solution till the final concentration is 0.01-0.02 percent, stirring (at the speed of more than or equal to 40 revolutions per minute) or homogenizing for 10 seconds, and preserving heat for 30 minutes; C, adding water in an amount of 1-5 percent based on the weight of the crude oil, and stirring uniformly; and D, adding enzyme in an amount of 0.005-0.015 percent (W / W) based on the weight of the crude oil, stirring and reacting for 1-3 hours, and centrifuging or naturally precipitating for separating degummed oil from a gum deposit. The method is practicable, and is easy and convenient to operate; when phosphatidase A2 is used for degumming, high-value enzyme hydrolyzed phospholipid can be obtained, so that energy consumption is lowered; and expressed (or leached) crude oil with very high phosphorus content or hydrated degummed oil with relatively low phosphorus content can be directly used for degumming in the method, so that the method has a wider application range.

The invention relates to a method for preparing high-emulsibility dried egg yolk, which relates to the method for preparing the dried egg yolk. The method solves the problems that the dried egg yolk prepared by the prior art has low emulsibility, poor thermal stability, and unsafety caused by introducing chemical radicals in the process of yolk chemical modification. The method comprises the following steps: firstly, washing fresh fowl eggs and separating out egg yolks, then mixing and stirring the egg yolks, water and CaCl2 to obtain an egg yolk solution, pouring the egg yolk solution into a reaction kettle, and adjusting the pH value of the egg yolk solution; secondly, adding phosphatidase into the reaction kettle to hydrolyze the egg yolk solution so as to obtain a hydrolysis solution; and thirdly, performing spray drying on the hydrolysis solution to obtain the high-emulsibility dried egg yolk. The dried egg yolk hydrolyzed and modified by the phosphatidase has stronger emulsibility and remarkably improved thermal stability, and no chemical radical is introduced in the preparation process so that the product is safe and reliable; the emulsifying activity of the dried egg yolk is improved by 30 percent compared with the prior dried egg yolk, and the emulsifying stability is 4 times of that of the prior dried egg yolk; and the thermal stability is good and the modifying temperature is between 85 and 100 DEG C.

The invention relates to a technique for preparing diacylglyceryl-phosphorylserine rich in unsaturated acid, which is characterized in that a reactor is preheated to 30 to 60 DEG C, squid lecithin and L-serine at the mass ratio of 1:1-15 is added, phosphatidase D2-15U / g is also added as reaction substrate, gas R134a is pumped to lead the pressure of the reactor to reach 4-6MPa, the pressure is reduced after agitating reaction for 2 to 8 hours, R134a is recycled after the reaction mixture comes into a separation pot which is communicated with the reactor under the temperature of 30 to 60 DEG C, reaction mixture is collected from the a sample receiving port of the separation pot, at last, the reaction mixture is washed by using acetone and centrifuged for 5 to 10 min to separate the solid phase for further purification. The diacylglyceryl-phosphorylserine prepared by the invention rich in high unsaturated fatty acid as the medium in catalytic reaction of phosphatidase D has the advantages of simple technique, low equipment input, short reaction time, high content of unsaturated fatty acid and high yield and the neglecting of organic solvent. Furthermore, R134a can be completely recycled after reaction, which is beneficial to the separation of products and repeated use of enzyme.

This invention open a cold tolerance of Serratia sand Habitat-chuen (Serratia fonticola) xjF1 CGMCC No.1971, through the strain characteristics, enzyme production conditions and the nature of enzyme research, proved a new phospholipase A1 gene consists of two open plA and plS reading frame, was certified plA gene coding phospholipase A1, plS gene coding phospholipase A1 auxiliary protein. This invention relates to the gene containing the recombinant plasmid and reorganization cell, the gene expression products phospholipase A1 in the show activity under low-temperature conditions, can be used in industries such as oil refining process can be simplified production process, lowering energy consumption, improve product quality , protecting the environment, reduce production costs, have important practical significance.

The invention discloses a processing method of soybean oil. The method comprises the following steps: 1, preparing crude soybean oil; 2, adding phosphatidase C, and carrying out primary degumming treatment to obtain primary degummed soybean oil; 3, adding phosphatidase A1, and carrying out secondary degumming treatment to obtain secondary degummed soybean oil; 4, carrying out alkali refining depickling treatment to obtain depickled soybean oil; 5, adding waste white clay to the depickled soybean oil, carrying out pre-decoloring treatment to obtain pre-decolored soybean oil, adding attapulgite and active carbon to the pre-decolored soybean oil, and carrying out re-decoloring treatment to obtain re-decolored soybean oil; and 6, carrying out deodorizing treatment on the re-decolored soybean oil to obtain finished soybean oil. The processing method of the soybean oil has the advantages of safety, environmental protection, simple process, convenient operation, low cost and high refining rate, and is suitable for soybean oil processing industrial production.

The invention relates to an acid soil amendment and its preparation method. the soil amendment contains the following ingredients (by weight): 4-12 parts of silicon dioxide, 1-6 parts of calcium oxide, 3-10 parts of biochar, 3-8 parts of lignin, 2-6 parts of urea, 2-6 parts of sodium silicoaluminate, 1-4 parts of organic bentonite, 0.3-0.7 part of polymaleic anhydride, 0.5-1 part of hydrogen peroxide, 0.3-0.7 part of beta-cyclodextrin, 0.4-1 part of phosphatidase, 0.2-0.8 part of powder of Chinese prickly ash and 0.2-0.8 part of sweet potato powder. The soil amendment provided by the invention can effectively raise pH value of acid soil and improve the defect of short period of soil. Meanwhile, the acid soil amendment can effectively kill noxious bacteria in soil and also can effectively reduce content of heavy metal ion in soil.

The invention relates to a method for microwave puffing pretreatment and water extraction of tea oil and other edible oil. The method comprises the following process steps of: adding ethanol at certain concentration, ammonia carbonate and other swelling agents, performing wet crushing, and performing microwave puffing treatment on tea seed and other oil; performing water extraction of oil, taking upper-layer oil, performing enzyme degumming by using phosphatidase, and filtering, or directly filtering; performing vacuum dehydration and drying to obtain the oil, dissolving tea saponin in tea seed residues in water, separating an aqueous phase, and clarifying and filtering to obtain an aqueous solution containing the tea saponin; and drying wet meal obtained through separation, and selling or developing into highly processed products such as tea seed sauce and the like. The production process has the characteristics of simple process, small investment and suitability for industrial production.

The invention provides a method for immobilizing phosphatidase A1. The method is improved on the technical basis of cross-linked enzyme aggregates and comprises the steps of: preparing free phosphatidase A1 into cross-linked phosphatidase aggregates; and then entrapping the obtained cross-linked phosphatidase aggregates to obtain immobilized phosphatidase A1. The influences of concentration of a phosphatidase liquid, saturation degree of a precipitating agent, precipitation Ph value, precipitation time, concentration of a cross-linking agent, cross-linking time, concentration of sodium alginate, concentration of calcium ions and other different immobilizing conditions on the immobilizing efficiency and the catalytic effect of the phosphatidase are researched, and the enzymology nature of an immobilized phosphatidase membrane is discussed. The advantage of immobilizing the phosphatidase by adopting the method for entrapping and cross-linking aggregates is further explained through an SEM (Scanning Electron Microscope) image of the immobilized phosphatidase membrane. The phosphatidase is immobilized by utilizing the excellent chemical stability, mechanical property and other properties of the entrapped and cross-linked phosphatidase aggregates, the activity and the stability of the phosphatidase in a reaction system are improved, the activity and the selectivity of the phosphatidase are adjusted and controlled, so as to be favorable for the recovery of the phosphatidase and the maintenance of higher activity recovery rate of the phosphatidase; and the activity recovery rate of the finally obtained immobilized phosphatidase is 80.2 percent, and the relative activity of the phosphatidase after being repeatedly used for seven times is kept at 65 percent above.

The invention relates to a method for preparing functional fatty, in particular to a preparing method of diacylglycerol. The preparing method comprises the steps that: (1) 50 shares of animal oil or vegetable oil in weight, 5-30 shares of water in weight and 0.03-0.20 shares of phosphatidase A1 in weight are mixed and reacted for 2 to 12 hours under the temperature of 30 to 70 DEG C and stand for 1 to 2 hours or centrifugalize to delaminate; (2) the oil layer gained by delamination on the condition of temperature of 80 to 95 DEG C and absolute pressure of 4000 to 6000 Pa is dried for 30 to 60 minutes to get crude diacylglycerol. Purified diacylglycerol can be gained through the elimination of free fatty acids under the temperature of 120 to 160 DEG C and absolute pressure of 0.1 to 40 Pa. Phosphatidase A1 used by the invention has fine catalysis effect and the price is only 5 percent to 10 percent of fatty acid enzyme which leads to significant decrease of cost and fine application prospect in industry.

The invention relates to a non-bitter fishy smell, instant whole egg powder and a preparation method thereof. The method comprises: (a) adopting two-stage dynamic high-pressure micro-jet treatment; (b) sequentially adding compound protease and phospholipid to the whole egg liquid Enzyme A1; wherein the composite protease is papain and flavor protease, the dosage ratio of the two is 2:1, the amount of composite protease added in the whole egg liquid is whole egg liquid: composite protease=100:0.8-1.2, add The amount is calculated by mass ratio; phospholipase A1 is enzymatically hydrolyzed at room temperature, and the added amount in the whole egg liquid is whole egg liquid: phospholipase A1=100:0.01‑0.05, and the added amount is calculated by kg / L; (c) After spray drying, whole egg powder is fluidized bed granulated with a binder. The invention cooperates with dynamic high-pressure micro-jet homogenization, enzymatic hydrolysis and fluidized bed technology to obtain whole egg powder with good instant solubility, and no stratification is found within 10 hours after brewing.

The invention relates to a phosphatidase A1 immobilization method. The method comprises the following steps: 1, allowing a carrier selected from macroporous adsorption resin or ion exchange resin to contact with phospholipase A1 in order to immobilize the phospholipase A1 on the carrier; and 2, drying the phospholipase A1 immobilized carrier obtained in step 1 in order to obtain immobilized phospholipase A1. The invention also provides immobilized phospholipase A1 prepared through the method, and an application of the immobilized phospholipase A1. The immobilized phospholipase A1 has the functions of esterification and acid hydrolysis, can be repeatedly used in the oil industry, and has the advantages of high reaction efficiency low cost and potential industrial application.

The present invention relates to a method for preparing hydrolyzed phospholipids by utilizing ultrasonic treatment and enzymic hydrolysis modification, belonging to the field of phospholipids modification technology. Said invention uses soybean hydrous oil foot as raw material, uses the phosphatidase A2 as enzyme for enzymic hydrolysis and adopts ultrasonic treatment method to make the soybean hydrous oil foot be modified in water phase so as to prepare the hydrolyzed phospholipids.

The invention discloses a high value utilization method of plant oil hydrated oil foots, which comprises the following steps: 1)adjusting temperature of oil foots hydration to 40-65 DEG C, adding lipase or phosphatidase, stirring and reacting until an acetone insoluble substance in a system enable 20-45% of weight reduction; and 2)centrifuging or standing and separating to obtain a light phase taking oil as a main component and a heavy phase containing modified phosphatide. The method makes appropriate hydrolysis on hydrated oil foots by lipase or phosphatidase, then through separation, the light phase taking oil as the main component and the heavy phase containing modified phosphatide can be obtained, and oil and modified phosphatide have high value. The method can realize high value utilization of plant oil hydrated oil foots.

The invention discloses a degumming method for camellia oil, which comprises the following steps: heating primary camellia oil to between 40 and 70 DEG C, and adjusting the pH value of the primary camellia oil to between 3.0 and 6.5; adding phosphatidase and stirring the mixture under heat insulation condition; and finally filtering or centrifugally separating the mixture. The camellia oil degummed by the method has low content of phospholipid, and the camellia oil degumming efficiency is high; and the method has simple process, mild reaction conditions and easy control.

The invention discloses a method for preparing diglyceride from phosphatidase A1 (Lecitase Ultra) through catalytic esterification, which comprises the following steps: (1) obtaining substrates after mixing fatty acid and glycerin according to a mol ratio of 0.5 / 1 to 2.5 / 1; and sequentially adding phosphatidase A1 and de-ionized water into the substrates to obtain reactant, wherein the addition of the phosphatidase A1 meets the requirement of adding 50 to 120 U phosphatidase A1 into each gram of the substrates, and the addition of the de-ionized water accounts for 4.0 to 12.0 wt percent of the mass of the substrates; (2) controlling the temperature between 30 and 55 DEG C under the condition that the reactant is in the vacuum, stirring the materials for reaction for 4 to 16h to obtain reaction liquid; and carrying out centrifugation delamination on the reaction liquid to obtain coarse diglyceride at the upper layer; and (3) carrying out molecular distillation on the coarse diglyceride under the conditions of the temperature between 160 and 165 DEG C and the pressure between 0.3 and 1.0 Pa to remove unreacted fatty acid and obtain the diglyceride.

The invention relates to actinomycete for production of phospholipase D and application thereof and belongs to the field of biotechnology. The phospholipase D produced by the actinomycetes after fermentation has catalytic action on soybean phosphatidylcholine and L-serine to efficiently produce phosphatidylserine through transesterification. The actinomycete is collected from a wellhead NO.1066 in Dongfeng town, Dawa County, Panjin city, Liaoning province, and is identified as Streptomycesbottropensis, preserved in the key laboratory for screening of new drugs in Liaoning province under the number of SYP-A-6632, and is preserved in CGMCC under the number of CGMCC No.6993. A screening method of the Streptomycesbottropensis includes: 1, activating strains to be activated; 2, preparing a screening plate; 3, screening and decomposing lecithin strains by a plate method; 4, detecting transesterification activity. The Streptomycesbottropensis has the advantages that the operation process is simple, implementation conditions are mild, phosphatidylserine can be bio-converted efficiently and quickly, and basis of industrial production of phosphatidylserine in future is laid.

The invention belongs to the technical field of genetic engineering of enzymes, and particularly relates to a method for obtaining phosphatidase B mutants with improved enzyme activity through a PCR (sequential error-prone) technology in vitro directed evolution, using the phosphatidase B mutants to prepare glycerophosphatidylcholine and performing oil degumming. The phosphatidase B mutant PLBM is obtained; the enzyme activity is improved by 25 percent through being compared with that of wild phosphatidase B.

The invention discloses a lipoprotein phosphatidase A2 detection reagent and a preparation method and a use method thereof. The detection reagent comprises the following components: an 20-200 mmol / L HEPES (hydroxyethyl piperazine ethanesulfonic acid) buffer solution, a 20-100mmol / L citrate buffer solution, a 50 mmol / L 1-myristoyl-2-(4-p-nitrophenol succinic anhydride)phosphatidylcholine solution, a 20-100U / L lipoprotein phospholipase A2 calibration product, and a 20-100U / L lipoprotein phospholipase A2 quality control product. The detection reagent can realize the detection of the lipoprotein phosphatidase A2 by a full automatic biochemical analyzer, can determine the content of the lipoprotein phosphatidase A2 in biological samples in a high-throughput, rapid and accurate mode, and has the advantages of easy operation, high sensitivity, high specificity, accurate results, etc.

The invention discloses a purpose of phosphatidase Dzeta2 protein on the aspect of adjusting the sensibility of plants to somatotropic hormone or the response aspect to the low phosphorus stress. The invention also discloses a preparation method for the plants which have high sensibility to the somatotropic hormone or high responsibility to the low phosphorus stress. The invention leads the phosphatidase Dzeta2 protein, and the somatotropic hormone sensibility and the low phosphorus stress responsibility of the plants to be related, genetically modified plants which have high sensibility to the somatotropic hormone or high responsibility to the low phosphorus stress can be prepared, thereby the application amount of field somatotropic hormone can be greatly decreased, the absorption of the plants to low phosphorus is increased, and the agricultural cost can be greatly reduced.

The invention relates to a vegetable fat enzymatic degumming method, which belongs to the technical field of vegetable fat degumming. The method in the invention comprises the following steps of: firstly, culturing corresponding phosphatidase in a seed culture medium and a soyabean lecithin-containing fermentation culture medium respectively by using microbes capable of generating the phosphatidase; secondly, adding the phosphatidase into soybean crude oil which is treated by acid and specifically hydrolyzing the phosphatide in the vegetable fat under the action of an enzyme to generate corresponding lysophosphatide and fatty acid; and thirdly, removing generated lysophosphatide and fatty acid by hydration. In the invention, the microbial enzyme is adopted as a catalyst, so that degumming efficiency is high; and the phosphatidase resource is wide, fermentation conditions are simple, process conditions are simple, and cost is low.

The present invention belongs to the field of food processing technology. The production process of high protein nutritious bread containing defatted walnut powder includes the following steps: dissolving phosphatidase in water to compound enzyme solution and activating; stirring wheat flour, defatted walnut powder, sugar, milk powder, salt, yeast and improver slowly in a dough mixer; adding water via stirring; adding phosphatidase solution and stirring in gradually increased speed to obtain tough dough; adding butter while stirring in changed speed until the dough can form thin sheet; relaxing, cutting, forming, leavening and roasting. Adding defatted walnut powder into the bread material can raise the protein content in bread and increase the nutritious matter and fragrancy; and adding phosphatidase can increase the specific volume of bread and improve the texture and taste of bread.

The invention provides a preparation method for single cream hydrolysate. The preparation method comprise a step of subjecting homogenized single cream to enzymatic hydrolysis, wherein hydrolase for enzymatic hydrolysis comprises lipase and phosphatidase, and the activity units of lipase and phosphatidase for hydrolysis of each 100 kg of the single cream are 2.3 * 10<5> to 2.3 * 10<7>, and 5.25 * 10<5> to 5.25 * 10<7>, respectively. Through combined action of lipase and phosphatidase, an enzymatic hydrolysis speed is increased, and output efficiency of flavor components is high. The invention further provides a dairy product prepared from the single cream hydrolysate and a preparation method thereof.

The invention provides a split cladosporium-expressed phosphatidase C strain, and is preserved in General Microbiological Center of China Committee of Culture Collection for Microorganisms with a preservation number of CGMCC No. 7508. The provided bacterial strain and phosphatidase C prepared by the preparation method are used for grease refining, degumming effect is good, and the bacterial strain and phosphatidase C prepared by the preparation method can be used for grease refining, additive and medicine fields.

The invention provides phosphatidase, a coding gene, a preparation method and application thereof. Specifically, the invention provides isolated polypeptide, which contains: (1) the 21st to 565th amino acid sequence of SEQ ID NO:2; or (2) a polypeptide derived from (1) after substitution, deletion or addition of one or more amino acids in the amino acid sequence of (1) and reservation of the 21st to 565th amino acid sequence of SEQ ID NO:2. The invention also relates to a polynucleotide sequence for coding the polypeptide, a nucleic acid construct containing the polynucleotide sequence, a host cell of the polynucleotide sequence or the nucleic acid construct, a composition containing the polypeptide, and application of the polypeptide.

A method, and associated kit, for assaying the activity of lysosomal enzymes present in dried bodily fluids and cell tissue samples, such as α-L-iduronidase, β-D-galactosidase, β-D-glucosidase, chitotriosidase, total α-D-galactosidase and α-D-galactosidase A, hexosaminidase A and B, α-D-mannosidase, β-D-mannosidase, α-L-fucosidase, N-acetyl-α-galactosaminidase, arylsulfatases, sphingomyelinase, β-galactocerebrosidase, iduronate-2-sulfatase and β-D-glucuronidase. The method includes: (a) combining with a dried bodily fluid or cell tissue sample containing at least one type of lysosomal enzyme: (1) an eluent, (2) an incubation buffer and (3) a substrate or substrates capable of reacting with the assayed lysosomal enzymes and producing their corresponding enzyme product or products, (b) allowing the dried bodily fluid or cell tissue sample to react with the eluent, incubation buffer and substrate or substrates for an adequate time and temperature, and (c) applying measuring means to the enzyme product to determine the activities of the lysosomal enzymes present.

The invention discloses a freeze-drying protective agent for sphingomyelinase and a preparation method, and belongs to the technical field of biology. The freeze-drying protective agent of sphingomyelinase comprises the following components: 0.2%-0.3% (w / v) of cane sugar, 0.1%-0.4% (w / v) of trehalose, 0.2%-0.5% (w / v) of D-mannitol, 0.1%-0.4% (w / v) of glycine, 0.02%-0.05% (v / v) of Tween 20 and 5-10 mM of Mg<2+>. Through formula screening and optimization, the obtained sphingomyelinase freeze-dried powder has the characteristics of high enzyme activity, good thermal stability, long storage time and good appearance.

A gene of the thermophilic lipase / phosphatidase A2, the engineering bacterium expressed by its recombinant vector in host, the thermophilic lipase / phosphatidase constructed with said engineering bacterium and the industrial use of said enzyme are disclosed. Said enzyme A2 is prepared from the thermophilic Aeropyrum pernix K1 through constructing engineering bacterium, fermenting, centrifugal separation, collecting deposit, freeze thawing, ultrasonic breaking, heat treating, and affinity chromatography.