CN101153311B - Method of producing vegetable seed oligopeptide and product thereof - Google Patents
Method of producing vegetable seed oligopeptide and product thereof Download PDFInfo
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Abstract
The invention belongs to the natural material extraction field, and relates to the method to produce oligopeptide of rape seeds from rape protein, as well as the product thereof. The procedures includes that: the raw material is grinded and sifted by a 60-80-hole screen, then 100 weight potions of the sample is taken, added with 1,000-2,000 weight potions of water, soaked inside a reaction pot under 20-25 DEG C for 1-2 hours, and the pH is regulated to 9-12 with 1-4mol.L(-1) NaOH, added with 1-5 weight potions of alkali proteinase, and undergoes enzymatic hydrolysis for 1-5 hours under 3-50 DEG C, then the pH of the hydrolysate liquid is regulated with 1-4mol.L(-1) HCl to 5-7, and the liquid undergoes enzyme inactivation for 5-10 minutes, before being centrifuged under 4000r.min(-1) to obtain the supernate 1, the pH of which is regulated to 2-4 with 1-4mol.L(-1) HCl, and undergoes decoloration for 1-3 hours after 0.5-1.5 weight potions of the active carbon are added, then centrifuge is carried out under 4000r.min(-1) followed by filtration to obtain the supernate 2, then the separated liquid produced through nanometer filtration/separation method is refrigerated and dried in vacuum, to obtain the component of oligopeptide of rape seeds with the molecular weight of 1,000-800, 800-600, 600-400, and 400-200 daltons, then the permeated final liquid is treated by reverse infiltration method to obtain the cycling water in the procedures. The oligopeptide of rape seeds produced by the invention as the raw material for healthcare product needs further exploitation.
Description
Technical field
The invention belongs to natural product extraction and health food processing technology.Be specifically related to from the rape protein raw materials, prepare the method and the product of vegetable seed oligopeptide (biologically active peptides); The be otherwise known as level of little peptide, oligopeptides or oligopeptide of this biologically active peptides is divided product; Its range of molecular weight distributions is 1000～800,800～600,600～400; 400～200 dalton the invention still further relates to the vegetable seed oligopeptide product by method for preparing.
Background technology
Rape seed protein is a kind of quality protein source, and its amino acid bio PER value of tiring is close with the WHO/FAO recommendation.Yet because it contains ANFs such as sulphur glucoside, tannin, phytic acid, limiting it can only feeding or as fertilizer sources, and can not be used for food.China is rape big producing country; Grouts behind the annual only vegetable seed system oil are just up to 600～7,000,000 tons; If can contained protein wherein be fully utilized, developing is of value to the functional factor of extraordinary crowd's needs food, will produce huge economic and social benefit.In recent years, the various countries scholar makes remarkable progress in the research aspect the vegetable seed detoxification: the content that has effectively reduced poisonous sulphur glucoside thing and erucic acid like breeding of double-low rapeseed; Physics, chemistry and biochemical methods such as organic solvent extraction, fermentation neutralization, extruding puffing all have positive effect to the content that reduces polyphenol, phytic acid etc. in the vegetable seed.The restrictive factor that influences the rape seed protein comprehensive utilization just progressively obtains removing, and its edible prospect is generally good.
Because antitumor, anti-oxidant, multiple functional property such as prevention cardiovascular and cerebrovascular diseases etc. that rapeseed peptide (rsp) has, become the research focus of present exploitation rape seed protein.Relevant document shows that the preparation of rapeseed peptide (rsp) mainly concentrates on uses enzymatic hydrolysis, and obtains various rapeseed peptide (rsp) levels through gel chromatography and ion exchange resin separation and purification and divide product.Existing deficiency is that to be difficult to obtain molecular weight through aforesaid method be that 1000～200 molecular weight are 1000～800,800～600, the product between 600～400,400～200 dalton (Da), and its biological activity of the product between these molecular weight areas is best; Because the gel chromatography technology is unfavorable for suitability for industrialized production, contain a certain amount of amino acid in the currently available products again, influenced the performance of little peptide biological activity to a certain extent.Number of patent application 200610019136 documents disclose a kind of preparation method of rapeseed peptide (rsp); The gained polypeptide liquid is decoloured through charcoal absorption; The refined liquid after the ion exchange resin desalination and the good classification of being unrealized still are the mixed solution of multiple peptide, and the molecular-weight gradation scope is indeterminate; The document of number of patent application 02133461.7 and number of patent application 02133463.3 discloses the method that adopts cross-flow ultrafiltration technology and ion exchange chromatography separation and purification polypeptide; Selection through ultra-filtration membrane can molecular weight cut-off 1000 molecular weight be 1000～800; 800～600,600～400, the activeconstituents below 400～200 dalton; On the molecular weight size, can reach the requirement of oligopeptide (biologically active peptides); But can't realize further classification through ultrafiltration, and the activity of oligopeptide is directly related with the molecular weight size with its structure, is necessary its further grading purification to this part bioactive peptide; Adopt ion-exchange chromatography pattern purifying, active peak is not concentrated, and the resin bad mechanical strength is prone to swelling and receives Organic pollutants, the tediously long technological deficiency such as time-consuming of steps such as regenerative operation.
Summary of the invention
The objective of the invention is to overcome the defective that prior art exists; Adopt enzyme engineering technology; R-o-and nanofiltration membrane separation technology; Provide a kind of vegetable seed oligopeptide level for preparing to divide the method for product and utilize the vegetable seed oligopeptide level of this method acquisition to divide product, utilizing this method to obtain molecular weight is that 1000～200 daltonian vegetable seed oligopeptide levels are divided product.
A kind of preparation method of vegetable seed oligopeptide: it is prepared by the following step:
(1) with proteinaceous Semen Brassicae campestris raw material or tentatively from Semen Brassicae campestris, extract proteic material sample and grind according to ordinary method, cross 60～80 mesh standard sieves, sample is subsequent use as handling;
(2) take by weighing processing sample 100 weight parts in the step (1), add water 1000～2000 weight parts, 20～25 ℃ are soaked 1～2h in 2～3L reaction kettle, obtain soak solution;
(3) with the soak solution of step (2) with 1～4molL
-1NaOH solution adjust pH to 9～12 add 1～5 weight part Sumizyme MP, and enzymolysis 1～5h under 30～50 ℃ of conditions obtains enzymolysis solution;
(4) with the enzymolysis solution of step (3) with 1～4molL
-1HCl solution adjust pH to 5～7, the enzyme 5～10min that under 90～100 ℃ of conditions, goes out obtains hydrolyzed solution;
(5) hydrolyzed solution of step (4) is put 4000rmin
-1The centrifugal down supernatant 1 that gets is with 1～4molL
-1HCl solution is transferred pH value to 2～4 of this supernatant 1, under 30～50 ℃ of conditions, adds 0.5～1.5 weight part gac, and decolouring 1～3h obtains hydrolyzed solution;
(6) with step (5) through the decolouring after hydrolyzed solution at 4000rmin
-1Down centrifugal, to cross and filter supernatant 2, the molecular weight that makes the supernatant 2 after the decolouring is less than more than 70% of 1000 daltonian ingredients constitute X 1000 total amounts;
(7) but to select molecular weight cut-off for use be 1000～800,800～600,600～400; 400～200 daltonian tubular fibre Nano filtering composite membranes carry out classification to the supernatant 2 of described step (6), and obtaining molecular weight is 1000～800,800～600; 600～400,400～200 daltonian grades of separatory, operational condition is: pressure is 0.6～0.8MPa; Temperature of reaction is 25～40 ℃, and control feedstock solution concentration is lower than 5%;
(8) separatory at different levels in the step (7) (are adopted vacuum freeze drying with vacuum freeze drying; Available from German Alpha Co., Ltd, instrument model: 1-4LD carries out according to the instrumentation specification sheets) handle; Obtaining molecular weight is 1000～800; 800～600,600～400,400～200 daltonian vegetable seed oligopeptide levels are divided product;
(9) be 1000～800 with step (7) through molecular weight cut-off; 800～600; 600～400; It is that 25～35 ℃, pH value of solution value are under 4～7 conditions in working pressure 300～370psi, temperature of reaction that 400～200 daltonian nf membrane ultimate sees through liquid, through the impurity and purification of polyamide reverse osmose membrane assembly, recycles as process water;
The present invention has following advantage:
(1) having obtained molecular weight through nanofiltration is 1000～800,800～600,600～400 at 1000 molecular weight; Vegetable seed level below 400～200 dalton is divided oligopeptide, and 400～200 molecular weight are 1000～800,800～600; 600～400; The MWD that 400～200 dalton's levels are divided peptide is the present minimum little peptide crowd of molecular weight section in the world, has extremely strong biological activity and variety.
(2) to adopt the characteristics of nanofiltration be the nf membrane combination that can hold back scope through different molecular weight in the present invention; Obtain the oligopeptide product of various molecular weights distribution range; Help having the exploitation of the functional product innovation of overall coordination effect, method is simple.
(3) rape seed protein enzymolysis and enzymolysis solution separation and purification are all accomplished 50 ℃ of following temperature of reaction, and the enzyme time of going out is short, help farthest keeping the activity of vegetable seed oligopeptide.
(4) degree of hydrolysis of rape seed protein enzymolysis solution of the present invention (DH) is controlled between 10～15%.The hydrophobic amino acid of this degree of hydrolysis scope occupy C, the N end of protein (or peptide) more, and the formation of the enzymolysis solution bitter taste that weakened needn't be adopted and add food flavor enzyme etc. and go bitter means, has reduced production cost.
Description of drawings
Accompanying drawing 1: the preparation technology's flow process that is vegetable seed oligopeptide of the present invention.
Accompanying drawing 2: be the gel filtration chromatography figure that characterizes MWD situation in the decolouring supernatant.
Embodiment
Below through specific embodiment the present invention is further specified; The embodiment that lifts is not the restriction to the present invention's protection; Just more describe in detail of the present invention, the person skilled in the art of association area can make the adjustment and the improvement of non-intrinsically safe property to some content of this invention.
Embodiment 1: be the feedstock production oligopeptide with the virus-free rapeseed meal
According to process step and the technical parameter that " summary of the invention " of this specification sheets front described, take by weighing that (preparation of virus-free rapeseed meal is referring to Zhao Hailing etc., dregs of rapeseed cake detoxification process Research on parameters through the virus-free rapeseed meal that grinds, cross 60 mesh standard sieves; Chinese oil, 2003,28 (12): 23～27) 100kg; Add clear water 2000kg; 20～25 ℃ are soaked 2h in the 3L reaction kettle, obtain soak solution, use 4molL
-1NaOH solution adjust pH to 11 adds 5kg Sumizyme MP 2709 (" honeybee " board is available from the magnificent prosperous and powerful Bioisystech Co., Ltd in east, Beijing), at 50 ℃ of following enzymolysis 4h of temperature of reaction, uses 1molL
-1HCl solution adjust pH to 5, the enzyme 10min that under 90 ℃, goes out obtains enzymolysis solution, and this enzymolysis solution is put 4000rmin
-1The centrifugal down supernatant 1 that gets is used 1molL
-1HCl solution is transferred the pH value to 4 of supernatant 1, under 50 ℃ of conditions, adds the 1.5kg activated carbon decolorizing 3 hours, 4000rmin
-1Centrifugal down, mistake filters supernatant 2.Adding water adjustment supernatant 2 concentration is 4%, starts HPP, and it is 1000～800,800～600 that feed liquid is squeezed into molecular weight cut-off 1000 molecular weight, and 600～400,400～200 daltonian tubular fibre Nano filtering composite membrane assemblies are handled.Working pressure is 0.8MPa, and temperature is 35 ℃, and feed rate is 1500L/h, sees through under the flow quantity 120L/h condition, and through the 120min circulation, detecting and seeing through liquid concentration is 2.5%, shut-down operation.Collect and see through liquid; Under above-mentioned operational condition; To carry out classification through the tubular fibre Nano filtering composite membrane assembly of 800Da, 600Da, 400Da, 200Da successively through liquid, reach shut-down operation in 45% o'clock, and collect the each several part liquid concentrator in each liquid concentrator concentration; Obtain the oligopeptide level separatory of MWD at 1000～800Da, 800～600Da, 600～400Da, 400～200Da, separatory protein contents at different levels account for respectively the enzyme hydrolyzate Tot Prot 5%, 10%, 15%, more than 30%.Separatory at different levels are through vacuum freeze drying (vacuum freeze drying; Available from German Alpha Co., Ltd; Instrument model: 1-4LD; According to instrumentation specification sheets operation) handle, obtain molecular weight and be respectively 1000～800,800～600,600～400 and 400～200 4 kinds of vegetable seed oligopeptide levels and divide product.The liquid that sees through through 200 dalton's nf membrane is that 360psi, temperature of reaction are that 35 ℃, pH value of solution value are under 4 the condition at working pressure, through the impurity and purification of polyamide reverse osmose membrane assembly, recycles as process water.In the present embodiment, the rapeseed protein degree of hydrolysis is 10～15%, and molecular weight accounts for more than 70% of X 1000 total amount less than the composition of 1000Da in the decolouring supernatant.The dry back of gained oligopeptide liquid cooling at different levels minutes freeze-drying appearance luster is light yellow. and Powdered, index protein contnt (N * 6.25, butt) % >=95; PH value 4～7, polyphenol content %＜O.5, phytic acid content %＜0.2; The sulphur glucoside must not detect, total free aminoacids %＜1.
Embodiment 2: be the feedstock production oligopeptide with the virus-free rapeseed meal
According to the process step of embodiment 1, take by weighing through grinding, cross virus-free rapeseed meal (Zhao Hailing etc., the dregs of rapeseed cake detoxification process Research on parameters of 70 mesh standard sieves; Chinese oil, 2003,28 (12): 23～27) 150kg; Add clear water 2000kg, 20～25 ℃ are soaked 2h in the 3L reaction kettle, use 3molL
-1NaOH solution adjust pH to 10 adds 3kg Sumizyme MP 2709 (" honeybee " board is available from the magnificent prosperous and powerful Bioisystech Co., Ltd in east, Beijing), and enzymolysis 3.5h under 45 ℃ of conditions of temperature obtains enzymolysis solution, uses 2molL
-1HCl solution is transferred this enzymolysis solution pH value to 4.5, the enzyme 10min that under 90 ℃ of conditions, goes out, 4000rmin
-1The centrifugal down supernatant 1 that gets.Use 2molL
-1HCl solution is transferred the pH value to 4 of supernatant 1, under 45 ℃ of conditions, adds 2kg activated carbon decolorizing 2.5h, 4000rmin
-1Centrifugal down, mistake filters supernatant 2.Adding water adjustment supernatant concentration is 3.5%, starts HPP, feed liquid is squeezed into the tubular fibre Nano filtering composite membrane assembly of molecular weight cut-off 1000Da and is handled.At working pressure 0.7MPa, 30 ℃ of temperature, feed rate 1200L/h sees through under the flow quantity 100L/h condition, and through the 150min circulation, detecting and seeing through liquid concentration is 3.0%, shut-down operation.Collect and see through liquid; Under above-mentioned operational condition; To carry out classification through the tubular fibre Nano filtering composite membrane assembly of 800Da, 600Da, 400Da, 200Da successively through liquid, reach shut-down operation in 40% o'clock, and collect the each several part liquid concentrator in each liquid concentrator concentration; Obtain the oligopeptide level separatory of MWD at 1000～800Da, 800～600Da, 600～400Da, 400～200Da, separatory protein contents at different levels account for respectively the enzyme hydrolyzate Tot Prot 5%, 15%, 15%, more than 25%.Vacuum freeze-drying method by embodiment 1 introduces is handled separatory at different levels, obtains 1000～800Da, 800～600Da, 600～400Da, four kinds of vegetable seed oligopeptides of 400～200Da level and divides product.Through the 200Da nf membrane see through liquid under working pressure 370psi, 35 ℃ of temperature, pH value of solution value 5 conditions, through the impurity and purification of polyamide reverse osmose membrane assembly, recycle as process water.In the present embodiment, the rapeseed protein degree of hydrolysis is 11～14%, and molecular weight accounts for more than 75% of X 1000 total amount less than the composition of 1000Da in the decolouring supernatant.The dry back of gained oligopeptide liquid cooling at different levels minutes freeze-drying appearance luster is light yellow, and is Powdered, index protein contnt (N * 6.25; Butt) % >=97, pH value 3.5～6.5, polyphenol content %＜0.4; Phytic acid content %＜0.1, the sulphur glucoside must not detect, total free aminoacids %＜0.5.
Embodiment 3: be the feedstock production oligopeptide with the rapeseed protein concentrate
Took by weighing 70 mesh sieves rapeseed protein concentrate (Wang Haiying, Wu Moucheng, Ceng Xiaobo, the acetone extraction legal system is got rapeseed protein concentrate, Chinese grain and oil journal; 2001,16 (4): 10～13) 100kg adds clear water 1500kg; Room temperature is soaked 1.5h for 20～25 ℃ in the 3L reaction kettle, uses 2molL
-1NaOH solution adjust pH to 10 adds 3kg papoid (Nanning Jie Woli biological products ltd), and enzymolysis 3h under 45 ℃ of conditions of temperature uses 2molL
-1HCl solution adjust pH to 6, the enzyme 8min that under 95 ℃ of conditions, goes out, 4000rmin
-1The centrifugal down supernatant that gets.Use 1molL
-1HCl solution is transferred supernatant pH value to 3, under 45 ℃ of conditions, adds 1.0kg activated carbon decolorizing 2h, 4000rmin
-1Centrifugal down, mistake filters supernatant.Adding water adjustment supernatant concentration is 5%, starts HPP, feed liquid is squeezed into the tubular fibre Nano filtering composite membrane assembly of molecular weight cut-off 1000Da and is handled.At working pressure 0.7MPa, 30 ℃ of temperature, feed rate 1800L/h sees through under the flow quantity 110L/h condition, and through the 150min circulation, detecting and seeing through liquid concentration is 3.0%, shut-down operation.Collect and see through liquid; Under above-mentioned operational condition; To carry out classification through the tubular fibre Nano filtering composite membrane assembly of 800Da, 600Da, 400Da, 200Da successively through liquid, reach shut-down operation in 40% o'clock, and collect the each several part liquid concentrator in each liquid concentrator concentration; Obtain the oligopeptide level separatory of MWD at 1000～800Da, 800～600Da, 600～400Da, 400～200Da, separatory protein contents at different levels account for respectively the enzyme hydrolyzate Tot Prot 5%, 10%, 15%, more than 30%.Vacuum freeze-drying method by embodiment 1 introduces is handled separatory at different levels, obtains 1000～800Da, 800～600Da, 600～400Da, four kinds of vegetable seed oligopeptides of 400～200Da level and divides product.Through the 200Da nf membrane see through liquid under working pressure 350psi, 30 ℃ of temperature, pH value of solution value 5 conditions, through the impurity and purification of polyamide reverse osmose membrane assembly, recycle as process water.In the present embodiment, the rapeseed protein degree of hydrolysis is 10～13%, and molecular weight accounts for more than 70% of X 1000 total amount less than the composition of 1000Da in the decolouring supernatant.The dry back of gained oligopeptide liquid cooling at different levels minutes freeze-drying appearance luster is light yellow, and is Powdered, index protein contnt (N * 6.25; Butt) % >=96, pH value 4～6, polyphenol content %＜0.3; Phytic acid content %＜0.15, the sulphur glucoside must not detect, and total free aminoacids is %＜0.8.
Embodiment 4: with the vegetable seed white protein is the feedstock production oligopeptide
Took by weighing vegetable seed white protein powder (Xue Zhaohui, Wu Moucheng, Yin Jingzhang, the Ruan Zheng of 80 mesh standard sieves; The preparation and the characteristic thereof of degreasing Huaza No.3 dregs of rapeseed cake protein isolates, Food science, 2004; 25 (4): 25～28) 100kg; Add clear water 1000kg, 20～25 ℃ are soaked 1h in the 2L reaction kettle, use 4molL
-1NaOH solution adjust pH to 10.5 adds 2kg stomach en-(available from Chongqing Lik-Sang thing difficult to understand pharmaceutical Co. Ltd), 1kg trypsin available from Chongqing Lik-Sang thing difficult to understand pharmaceutical Co. Ltd), enzymolysis 2h under 30 ℃ of conditions of temperature uses 4molL
-1HCl solution adjust pH to 6.8, the enzyme 5min that under 100 ℃ of conditions, goes out, 4000rmin
-1The centrifugal down supernatant 1 that gets.Use 4molL
-1HCl solution is transferred the pH value to 2.8 of supernatant 1, under 30 ℃ of conditions, adds 0.5kg activated carbon decolorizing 1h, 4000rmin
-1Centrifugal down, mistake filters supernatant 2.Adding water whole supernatant 2 concentration of withering is 3.5%, starts HPP, feed liquid is squeezed into the tubular fibre Nano filtering composite membrane assembly of molecular weight cut-off 1000Da and is handled.At working pressure is 0.65MPa, 25 ℃ of temperature, and feed rate 2000L/h sees through under the flow quantity 150L/h condition, and through the 100min circulation, detecting and seeing through liquid concentration is 2.0%, shut-down operation.Collect and see through liquid; Under above-mentioned operational condition; To carry out classification through the tubular fibre Nano filtering composite membrane assembly of 800Da, 600Da, 400Da, 200Da successively through liquid, reach shut-down operation in 35% o'clock, and collect the each several part liquid concentrator in each liquid concentrator concentration; Obtain the oligopeptide level separatory of MWD at 1000～800Da, 800～600Da, 600～400Da, 400～200Da, separatory protein contents at different levels account for respectively the enzyme hydrolyzate Tot Prot 5%, 5%, 20%, more than 25%.Vacuum freeze-drying method by embodiment 1 introduces is handled separatory at different levels, and four kinds of vegetable seed oligopeptides of the oligopeptide level separatory level that obtains molecular weight and be 1000～800Da, 800～600Da, 600～400Da, 400～200Da is divided product.Through the 200Da nf membrane see through liquid under working pressure 350psi, 25 ℃ of temperature, pH value of solution value 6 conditions, through the impurity and purification of polyamide reverse osmose membrane assembly, recycle as process water.In the present embodiment, the rapeseed protein degree of hydrolysis is 13～15%, and molecular weight accounts for more than 80% of X 1000 total amount less than the composition of 1000Da in the decolouring supernatant.The dry back of gained oligopeptide liquid cooling at different levels minutes freeze-drying appearance luster is light yellow, and is Powdered, index protein contnt (N * 6.25; Butt) % >=95, pH value 3～6, polyphenol content %＜0.4; Phytic acid content %＜0.2, the sulphur glucoside must not detect, total free aminoacids %＜0.6.
Embodiment 5: with rapeseed protein concentrate and vegetable seed protein isolates is the feedstock production oligopeptide
Take by weighing through grinding, excessively virus-free rapeseed meal (Zhao Hailing, Li Xuegang, He Guoju, the dregs of rapeseed cake detoxification process Research on parameters of the virus-free rapeseed meal of 60 mesh standard sieves, mistake 70 mesh standard sieves; Chinese oil, 2003,28 (12): 23～27) each 50kg; Stir, mix, add clear water 2000kg, 20～25 ℃ are soaked 1.5h in the 3L reaction kettle; Obtain soak solution, this soak solution is used 1molL
-1NaOH solution adjust pH to 11 adds 5kg2709 Sumizyme MP (" honeybee " board, the magnificent prosperous and powerful Bioisystech Co., Ltd in east, Beijing), and enzymolysis 1.5h under 45 ℃ of conditions of temperature obtains enzymolysis solution, uses 2molL
-1HCl solution is transferred the pH value to 6.5 of enzymolysis solution, the enzyme 5min that under 100 ℃ of conditions, goes out, 4000rmin
-1The centrifugal down supernatant 1 that gets.Use 2molL
-1HCl solution is transferred the pH value to 2.5 of supernatant 1, under 25 ℃ of conditions, adds 0.8kg activated carbon decolorizing 0.5h, 4000rmin
-1Centrifugal down, mistake filters supernatant 2.Adding water adjustment supernatant 2 concentration is 3.0%, starts HPP, feed liquid is squeezed into the tubular fibre Nano filtering composite membrane assembly of molecular weight cut-off 1000Da and is handled.At working pressure 0.60MPa, 25 ℃ of temperature, feed rate 1500L/h sees through under the flow quantity 120L/h condition, and through the 90min circulation, detecting and seeing through liquid concentration is 1.5%, shut-down operation.Collect and see through liquid; Under above-mentioned operational condition; To carry out classification through the tubular fibre Nano filtering composite membrane assembly of 800Da, 600Da, 400Da, 200Da successively through liquid, reach shut-down operation in 50% o'clock, and collect the each several part liquid concentrator in each liquid concentrator concentration; Obtain the oligopeptide level separatory of MWD at 1000～800Da, 800～600Da, 600～400Da, 400～200Da, separatory protein contents at different levels account for respectively the enzyme hydrolyzate Tot Prot 5%, 10%, 20%, more than 35%.Vacuum freeze-drying method by embodiment 1 introduces is handled separatory at different levels., obtain molecular weight and be 1000～800Da, 800～600Da, 600～400Da, four kinds of vegetable seed oligopeptides of 400～200Da level and divide product.Through the 200Da nf membrane see through liquid under working pressure 370psi, 25 ℃ of temperature, pH value of solution value 6.5 conditions, through the impurity and purification of polyamide reverse osmose membrane assembly, recycle as process water.In the present embodiment, the rapeseed protein degree of hydrolysis is 12～14%, and molecular weight accounts for more than 75% of X 1000 total amount less than the composition of 1000Da in the decolouring supernatant.The dry back of gained oligopeptide liquid cooling at different levels minutes freeze-drying appearance luster is light yellow, and is Powdered, index protein contnt (N * 6.25; Butt) % >=95, pH value 5～7, polyphenol content %＜0.5; Phytic acid content %＜0.1, the sulphur glucoside must not detect, total free aminoacids %＜1.
Claims (1)
1. the preparation method of a vegetable seed oligopeptide is characterized in that step is following:
Proteinaceous Semen Brassicae campestris raw material is ground, and it is subsequent use to cross 60～80 mesh standard sieves, obtains handling sample; Get described processing sample 100 weight parts, add water 1000～2000 weight parts, in reaction kettle, under 20～25 ℃, soak 1～2h, with 1～4molL
-1NaOH solution adjust pH to 9～12 add 1～5 weight part Sumizyme MP enzymolysis 1～4h under 30～50 ℃ of conditions, obtain enzymolysis solution, with this enzymolysis solution with 1～4molL
-1HCl solution adjust pH to 5～7, the enzyme 5～10min that under 90～100 ℃ of conditions, goes out obtains hydrolyzed solution, and this hydrolyzed solution is put 4000rmin
-1The centrifugal down supernatant 1 that gets is with 1～4molL
-1HCl solution is transferred pH value to 2～4 of this supernatant 1, under 30～50 ℃ of conditions, adds 0.5～1.5 weight part activated carbon decolorizing, 1～3h, 4000rmin
-1Centrifugal down, cross and filter supernatant 2; Adopt the nanofiltration stage division, using molecular weight cut-off respectively is 1000～800,800～600; 600～400,400～200 daltonian tubular fibre Nano filtering composite membrane assemblies carry out classification to described supernatant 2; Obtain vegetable seed oligopeptide level separatory; Separatory at different levels are carried out vacuum concentration and lyophilize, and to obtain molecular weight be 1000～800,800～600, and 600～400 and 400～200 daltonian vegetable seed oligopeptide levels are divided product; Adopt reverse-osmosis treated to purify the ultimate liquid that sees through of nanofiltration fractionated; Obtain process recirculated water, wherein: nanofiltration fractionated processing condition are: working pressure is 0.6～0.8MPa, and temperature of reaction is 25～40 ℃; Control feedstock solution concentration is lower than 5%; The processing condition of described reverse-osmosis treated are: working pressure 300-370psi, temperature of reaction is 25～35 ℃, transfers pH value to 4～7 of solution.
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| CN102524516A (en) * | 2012-02-24 | 2012-07-04 | 浙江省农业科学院 | Rapeseed polypeptide used for resisting inebriation and alleviating hangover and preparation method thereof |
| CN102643889B (en) * | 2012-04-28 | 2014-11-05 | 南京财经大学 | Antihypertensive active rapeseed peptide and preparation method and application thereof |
| CN102676621B (en) * | 2012-04-28 | 2014-03-19 | 南京财经大学 | Antihypertensive rapeseed peptide and preparation method and application of antihypertensive rapeseed peptide |
| CN104263786B (en) * | 2014-09-09 | 2017-06-06 | 江苏大学 | The method that stomach and intestine simulation digestion prepares rapeseed dregs protein antioxidant peptide liquid |
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