CN102994598A - Weak bitter corn oligopeptide with high content of alanine and leucine, and preparation method thereof - Google Patents
Weak bitter corn oligopeptide with high content of alanine and leucine, and preparation method thereof Download PDFInfo
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- CN102994598A CN102994598A CN2012104926166A CN201210492616A CN102994598A CN 102994598 A CN102994598 A CN 102994598A CN 2012104926166 A CN2012104926166 A CN 2012104926166A CN 201210492616 A CN201210492616 A CN 201210492616A CN 102994598 A CN102994598 A CN 102994598A
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Abstract
The invention belongs to the field of agricultural and sideline product processing and utilization technology with high added value, and discloses a weak bitter corn oligopeptide with high content of alanine and leucine, and a preparation method of the weak bitter corn oligopeptide. The method comprises the steps of: (1) dispersing and hydrating corn gluten meal; (2) carrying out gelatinization and alpha-high temperature amylase enzymolysis, degreasing and removing starch; (3) carrying out enzymolysis; (4) carrying out enzyme deactivation and decoloration; and (5) carrying out rough filtration, ultrafiltration and nanofiltration, and drying to obtain the weak bitter corn oligopeptide. The weak bitter corn oligopeptide contains 85.42-87.63% by mass of protein percent, 92.44-94.21% by mass of peptide with the relative molecular weight being less than 1000Da, 6.75-6.94% by mass of alanine, 14.12-14.66% by mass of leucine and 1.12-1.36% by mass of free amino acid. The weak bitter corn oligopeptide with high content of alanine and leucine can be widely used as raw material of health-care food and common food, and especially can be used in products used for sobering up and protecting the liver.
Description
Technical field
The invention belongs to agricultural byproducts high added value process and utilization technology field, be specifically related to high weak bitter taste maize oligopeptide of a kind of L-Ala and leucine content and preparation method thereof.
Background technology
Corn is one of Three major grain crops in the world, also is China Traditional Agricultural crop.2009, China's corn yield reached 1.64 hundred million tons, accounts for grain yield 20%, occupies the second place of the world.One of main application of corn is to produce W-Gum, the main byproduct that wet milling process is produced in the W-Gum process is Zein powder (Corn Gluten Meal, CGM), also claim bloom, Zein powder generally contains the protein about 60%, and main component is zein (68%), gluten (22%) and sphaeroprotein (1.2%).Contain hydrophobic amino acid and proline(Pro), the L-glutamic acid etc. such as a high proportion of leucine, Isoleucine, α-amino-isovaleric acid, L-Ala in the corn protein, but the essential amino acids content such as Methionin, tryptophane is lower, the amino acid of this uniqueness forms makes the zein poorly water-soluble, nutritive value is not high, has limited the deep processing and utilization of corn protein.For a long time, Zein powder discharges naturally mainly as the feed resource of livestock and poultry cheapness or with waste liquid.According to incompletely statistics, the annual Zein powder that naturally discharges with waste liquid in the whole nation has not only been wasted resource, and has been caused environmental pollution more than 80,000 tons.Therefore the deep processing of studying corn protein has important social and economic significance.
In recent years, research is found, the unbalancedness that zein amino acid forms is so that it has unique physiological function, can obtain to have the bioactive peptide of different physiological roles by scientific and reasonable biotechnology means, comprise absorption easy to digest, anti-oxidant, Ginseng Extract, reduce ammonia concentration, hypotensive, promote the physiologically active peptides such as alcohol metabolism, wherein promote alcohol metabolism effect aspect, because of maize oligopeptide and at present existing commercially available relieving the effect of alcohol, the medicine difference of sobering up, belong to environmental protective hygienic food, the mechanism of action is clear and definite, energy is relieving alcoholism and protecting liver rapidly, alleviate drunk bring uncomfortable, the chemical additives such as preservative-free, without traditional Chinese medicine ingredients, have no side effect, safe, have good market application foreground, the maize oligopeptide with the effect of sobering up has become the focus of domestic and international research and development.
Corn sobering-up peptide is to produce stable phosphopyridine nucleotide NAD+ by L-Ala and the leucine concentration that improves in the blood, strengthen the activity of ethanol dehydrogenase and acetaldehyde dehydrogenase, promote decomposition and the metabolism of ethanol in the body, thereby reduce the concentration of ethanol in blood, the liver damage that alcohol is caused can play prophylactic effect.So, can use L-Ala and leucic content, the L-Ala that particularly exists with the oligopeptide form, leucine content are weighed the sober up effect of corn sobering-up peptide.
The natural corn protein molecular is because effects such as hydrogen bonds, hydrophobic bond, disulfide linkage, and peptide chain is curled in protein molecular inside and is similar to spherically, and close structure and stronger hydrophobicity is arranged is difficult to by protease hydrolysis; In addition, also there are starch, grease and the pigment etc. of a great deal of in the maize yellow-powder, affect the deep processing and utilization of corn protein.At present, how efficiently corn protein deep processing aspect research both at home and abroad mainly concentrate on the enzymolysis zein, obtains the small molecular weight maize oligopeptide of high yield.The outstanding difficult problem of maize oligopeptide preparation comprises four aspects: (1) product purity is low, and namely the maize oligopeptide protein content is on the low side, mainly be early stage the Zein powder purification processes do and can not get the position, affected the quality of product; (2) preparation technology loaded down with trivial details, without scientific optimization, cost is high, is difficult to industrializing implementation; (3) the dense even stimulation of the mouthfeel bitter taste of corn functional peptides is difficult to be accepted by popular, applies being restricted; (4) maize oligopeptide has multiple physiological function, but not for a certain given efficacy research and development, product efficacy is not outstanding.Disclose take maize yellow-powder (about protein content 60%) as raw material such as patent CN101531700A, the employing buck is processed, except destarching, the impurity such as grease obtain high purity zein material slag, again with this material slag pre-treatment, enzymic hydrolysis, centrifuging, concentrate drying is prepared protein content more than 85%, the maize oligopeptide powder of molecular weight distribution below 2000Da, this invention neutral and alkali proteolytic enzyme consumption is 5%～8%, the neutral protein enzyme dosage is 5%～8%, total enzyme dosage reaches 10%～16%, water consumption, alkali, the acid amount is large, product yield (corn peptide and Zein powder mass ratio) 22.3%～24%, the manufacturing cost of product is very high; In addition in the patent not decolouring goes peculiar smell technique, do not describe product special flavour, also not specifically for a certain effect design of product; Patent CN102028093A discloses take maize yellow-powder as raw material, adopt Sumizyme MP Alcalase, compound protease Protamex subsection enzymolysis, enzymolysis product is through centrifugation, supernatant liquor is respectively through 100kDa, 10～6kDa blocks the ultra-filtration membrane ultrafiltration of molecular weight, again that filtered solution is concentrated, spraying drying obtains the corn sobering-up peptide powder, molecular weight accounts for 73.19% less than the peptide section of 1200Da in this product, L-Ala and leucic content are respectively greater than 6.43% and 12.42%, the corn peptide purity of product is low, the local flavor situation is not described, in addition because only removing macromolecular substance by roughing out and ultrafiltration in the invention, the product Determination of Free Amino Acids that obtains, ash oontent does not provide data, the actual L-Ala that exists with the oligopeptide form, leucine content is unclear; Patent CN102172293A discloses a kind of corn active small peptide sober-up agent preparation method, adopting buck to process Zein powder purifies, add again Sumizyme MP Alcalase4～5%, flavor protease Flavourzyme and compound protease Protamex1～3% graded hydrolysis, supernatant liquor is the little peptide of 200～1000Da through two-stage ultrafiltering membrane sepn acquisition main molecules amount after the enzymolysis product centrifugation, this method is large with the enzyme amount, reaches 5%～8%.
Summary of the invention
For difficult point and the weak point that solves above-mentioned prior art, primary and foremost purpose of the present invention is to provide the preparation method of the high weak bitter taste maize oligopeptide of a kind of L-Ala and leucine content.This preparation method adopts degreasing, destarch to process, and prepares L-Ala, weak bitter taste maize oligopeptide that leucine content is high in conjunction with enzymolysis, activated carbon selective adsorption, concentrated, drying, and is with low cost, is fit to suitability for industrialized production.
Another object of the present invention is to provide L-Ala and the high weak bitter taste maize oligopeptide of leucine content that obtains by above-mentioned preparation method.This peptide can be used as the raw material of protective foods, bread and cheese, especially can be used for the Soboring-up liver-protecting product.
The present invention realizes by following technical proposals: the preparation method of the weak bitter taste maize oligopeptide that a kind of L-Ala and leucine content are high may further comprise the steps:
(1) dispersion of Zein powder, aquation: Zein powder is pulverized, be formulated as corn protein solution with deionized water, with corn protein solution dispersion, aquation;
(2) gelatinization, degreasing, destarching: the pH value of the corn protein solution that regulate to disperse, aquation is good is 5～7, is heated to 80 ℃～85 ℃, and maintenance 20min～60min, so that the zein molecular structure fully launches, and makes wherein the abundant gelatinization of starch; Add the α-alpha-amylase that is equivalent to Zein powder quality 0.1%～1%, under 80 ℃～85 ℃ conditions, behind stirring enzymolysis 30min～60min, passivation α-high-temperature starch enzymic activity, cooling down, centrifugal, degrease and liquid are got precipitation; Precipitation is the purification zein behind degreasing and the destarching;
(3) enzymolysis: the precipitation dispersed with stirring that step (2) is obtained with deionized water is the zein slurry, the mass percent that guarantees protein in the zein slurry is 5%～20%, the pH value of adjusting corn protein milk is 6～7, add the neutral protease that is equivalent to protein quality 0.5%～2% in the zein slurry and stir hydrolysis 1h～2h in 50 ℃～55 ℃, add again the aspartic protease be equivalent to protein quality 0.5%～2% in the zein slurry and stir hydrolysis 4h～8h in 50 ℃～55 ℃, finish to enzymolysis;
(4) go out enzyme, decolouring: after enzymolysis finishes, heat up, add the active carbon powder stirring and evenly mixing, constant temperature stirs 30～60min, realizes going out enzyme, decolouring, lifting L-Ala and leucic content;
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slurry coarse filtration after the enzyme that will go out, the decolouring, clear liquid after the press filtration is by ultrafiltration, and filtered solution passes through nanofiltration again, finishes desalination, removes total free aminoacids and concentrated, drying obtains the high weak bitter taste maize oligopeptide of L-Ala and leucine content.
Zein powder described in the step (1) (claiming again bloom) requires to reach at least feed and stipulates with Zein powder NYT685-2003, preferred one-level feed Zein powder;
The mass percent of protein is more than 60% in the described Zein powder;
Described pulverizing refers to Zein powder is crushed to 80 orders;
The mass percent of the final concentration of Zein powder is 5%～20% in the described corn protein solution;
The velocity of shear of described dispersion, aquation is 2000rpm～3000rpm, and the time is 10 ~ 20min.
PH value described in the step (2) is by using sodium hydroxide solution to regulate;
The concentration of described sodium hydroxide solution is 1mol.L
-1～5mol.L
-1
The stirring velocity of described stirring enzymolysis is 36rpm;
Described passivation α-high-temperature starch enzymic activity refers to enzymolysis solution is kept 10min～20min at 95 ℃～100 ℃;
Described cooling down refers to be cooled to 55 ℃～60 ℃;
Described centrifugal be centrifugal 10min～20min under 4000rpm～6000rpm.
The temperature of the deionized water described in the step (3) is 50 ℃～55 ℃.
Intensification described in the step (4) refers to that temperature rises to 80 ℃～85 ℃;
The add-on of described active carbon powder is equivalent to 0.2%～1% of protein quality in the zein slurry; Used gac is the above gacs of 200 orders.
Coarse filtration described in the step (5) refers to the zein stock pump after the enzyme that goes out, the decolouring is entered plate and frame diatomite filter coarse filtration;
Described ultrafiltration refers to the first ultra-filtration membrane ultrafiltration by molecular weight cut-off 100kDa of the stillness of night after the press filtration, and filtered solution is the ultra-filtration membrane ultrafiltration by molecular weight cut-off 5kDa again;
Described nanofiltration refers to that filtered solution is the nanofiltration membrane nanofiltration of 1nm through the aperture;
Described drying is preferably dry by drying tower, and it is 160 ℃～170 ℃ that operating parameters is preferably inlet temperature, and temperature out is 70 ℃～90 ℃.
The weak bitter taste maize oligopeptide that a kind of L-Ala and leucine content are high, prepared by aforesaid method, the mass percent of protein is 85.42%～87.63% in the preparation-obtained maize oligopeptide, relative molecular weight is 92.44%～94.21% less than the mass percent of 1000Da peptide, the mass percent of L-Ala is 6.75%～6.94%, and leucic mass percent is 14.12%～14.66%; The mass percent of total free aminoacids is 1.12%～1.36%, and this maize oligopeptide can be widely used as the raw material of protective foods, bread and cheese, especially can be used for preparing the Soboring-up liver-protecting product.
The present invention compared with prior art has following advantage and beneficial effect:
(1) the present invention adopts high temperature gelatinization, α-high-temperature starch enzyme liberating, centrifugal separation process, efficiently remove on the one hand starch, fat in the Zein powder, promote the purity of enzymolysis substrate Zein powder, continuous high temperature is processed the imporosity that can effectively open zein on the other hand, reduce its hydrophobicity, be convenient to follow-up enzymolysis, improve corn peptide product yield (relative molecular weight is 92.44%～94.21% less than the mass percent of 1000Da peptide), this method energy consumption is low, easy to operate, be easy to realization of industrialization.The imporosity that zein is opened at present research generally adopts the techniques such as high temperature high pressure process (processing more than the 10min such as about 121 ℃), micronizing, ultrasonication, or employing denaturing agent such as S-WAT etc., the former is such as patent CN101126103A, CN101411494A etc., and energy consumption is high, high to equipment requirements; The latter such as patent CN1786183A etc., it is residual that the denaturing agent of interpolation can produce, and reduces the security of product.The general alkali lye that adopts of starch, fat in the at present research removal Zein powder is processed centrifugal again high temperature or low-temperature amylase processing centrifuging process, such as patent CN101531700A, CN102172293A etc., this method has been removed grease in the alkali lye processing is centrifugal, simultaneously also lose the soluble proteins in the Zein powder, reduced the product yield; Process again starch technique after also having report to adopt the organic solvent extraction grease, have complex operation, residual safety issue.
(2) adopt to regulate initial slurry pH value to 6.0～7.0, neutral protease enzymolysis in the enzymolysis process of the present invention and cause that zein slurry pH value is down to the aspartic protease of adding below 5.0 enzymolysis, combination gradient enzymolysis can be given full play to the Action advantage of enzyme combination, improve enzymolysis efficiency and product yield, enzyme dosage 1～4% gets final product, and avoid introducing too much acid solution, alkali lye, production cost reduces greatly.In addition, the present invention adopts neutral protease and aspartic protease combination enzymolysis, has avoided adopting alkaline enzyme enzymolysis in most of bibliographical information, can significantly reduce the bitter taste of enzymolysis product, improves product special flavour.
(3) the present invention reaches decolouring, removes peculiar smell by the preferred activated carbon of science and absorbing process control selective adsorption coloring matter and bitter substance, this activated carbon has containing L-Ala, the low characteristics of leucic oligopeptide adsorption rate simultaneously, can obviously promote L-Ala, the content of leucine oligopeptide in product, the Soboring-up liver-protecting effect of improving product also is highly susceptible to industrialization.
The present invention is intended to take Zein powder (bloom) as raw material, adopt degreasing, destarch is processed, in conjunction with the gradient enzymolysis, the activated carbon selective adsorption, concentrated, drying is prepared the maize oligopeptide of weak bitter taste, this peptide protein matter content is more than 85%, relative molecular weight accounts for more than 90% less than the 1000Da oligopeptide, free aminoacid content is lower than 1.5%, a little less than the bitter taste, feature amino acid (L-Ala, leucine) content is high, the L-Ala that contains in the oligopeptide, leucine content reaches respectively 6.75%, more than 14.1%, can be widely used as protective foods, the raw material of bread and cheese especially can be used for the Soboring-up liver-protecting product.The present invention preferably realizes the preparation of industrialization maize oligopeptide of very economical property by the science of pre-treatment, enzymolysis, process for refining, superior product quality, bitter taste is weak, be easy to and other food ingredientss collocation, the Soboring-up liver-protecting effect is outstanding, can be used as the high-quality functional material and is widely used in varieties of food items, the healthcare products.The present invention can realize the high added value comprehensive utilization of precision corn deep processing by product, promotes precision corn deep processing Business Economic Benefit, promotes the benign cycle of corn planting, processing, cultivation.
Embodiment
The present invention is described in further detail below in conjunction with embodiment, but embodiments of the present invention are not limited to this.
Embodiment 1
(1) dispersion of Zein powder, aquation: with Zein powder (protein content 〉=60%, be purchased from Shandong west Wang Jituan) be crushed to 80 orders, with deionized water Zein powder being mixed with mass percent is 10% corn protein solution, with stirrer with the rotating speed of 2000rpm with Zein powder disperse, aquation 20min;
(2) gelatinization, degreasing, destarching: be 3mol.L with volumetric molar concentration
-1Sodium hydroxide solution the Zein powder pH value of solution is adjusted to 6.0, be heated to 82 ℃, and keep 40min so that the zein molecular structure fully launches, and make wherein the abundant gelatinization of starch; Add 0.5% α-alpha-amylase (the SUKAMy-Hi LQ20 be equivalent to the Zein powder quality, Su Kehan biotechnology company limited), enzymolysis 45min under 36rpm, 82 ℃ of conditions, after enzymolysis finished, enzymolysis solution kept 15min, the activity of α-passivation alpha-amylase in 95 ℃, then cooling down to 60 ℃, adopt separating centrifuge with the centrifugal 15min of 5000r/min rotating speed, the purification Zein powder after degrease and liquid, solid slag are degreasing and remove starch;
(3) enzymolysis: with 55 ℃ of deionized waters the solid slag dispersed with stirring is become the zein slurry, guarantee that protein content is 10% in the slurries; Regulate slurry pH value to 6.5, under 55 ℃ of constant temperature, add the neutral protease (Neutrase0.8L that is equivalent to protein quality 1% in the zein slurry, letter (China) Investment Co., Ltd of Novi) behind the stirring hydrolysis 1.5h, add again the aspartic protease be equivalent to protein quality 1% in the zein slurry (outstanding can the food-grade acid protease P rotex26L of section, the Wuxi is outstanding can biotechnology company limited of section) and stir hydrolysis 6h;
(4) go out enzyme, decolouring: enzymolysis is warming up to 85 ℃ after finishing, add 0.6% activated carbon powder (200 order) stirring and evenly mixing be equivalent to protein wt in the zein slurries, constant temperature 36rpm stirs 45min, contains L-Ala and leucic oligopeptide content in the enzyme of realizing going out, adsorption bleaching and the lifting product;
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slush pump that will contain activated carbon enters plate and frame diatomite filter coarse filtration, clear liquid after the press filtration at first is the ultra-filtration membrane ultrafiltration of 100kDa by molecular weight cut-off, filtered solution passes through the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa again, filtered solution is the nanofiltration membrane nanofiltration of 1nm again through the aperture, finish desalination, remove total free aminoacids and concentrated, concentrated solution drying tower is dry, inlet temperature is controlled to be 170 ℃, temperature out is controlled to be 80 ℃, the dry end, packing namely obtains the high weak bitter taste maize oligopeptide of L-Ala and leucine content.
Embodiment 2
(1) dispersion of Zein powder, aquation: with Zein powder (protein content 〉=60%, be purchased from Shandong west Wang Jituan) be crushed to 80 orders, with deionized water Zein powder being mixed with mass percent is 20% corn protein solution, with stirrer with the rotating speed of 3000rpm with Zein powder disperse, aquation 10min;
(2) gelatinization, degreasing, destarching: be 5mol.L with volumetric molar concentration
-1Sodium hydroxide solution the Zein powder pH value of solution is adjusted to 7.0, be heated to 80 ℃, and keep 60min so that the zein molecular structure fully launches, and make wherein the abundant gelatinization of starch; Add 1% α-alpha-amylase (the SUKAMy-Hi LQ20 be equivalent to the Zein powder quality, Su Kehan biotechnology company limited), enzymolysis 60min under 36rpm, 80 ℃ of conditions, after enzymolysis finished, enzymolysis solution kept 20min, the activity of α-passivation alpha-amylase in 95 ℃, then cooling down to 55 ℃, adopt separating centrifuge with the centrifugal 10min of 6000r/min rotating speed, the purification Zein powder after degrease and liquid, solid slag are degreasing and remove starch;
(3) enzymolysis: with 50 ℃ of deionized waters the solid slag dispersed with stirring is become the zein slurry, guarantee that protein content is 20% in the slurries; Regulate slurry pH value to 7.0, under 50 ℃ of constant temperature, add the neutral protease (Neutrase0.8L that is equivalent to protein quality 2% in the zein slurry, letter (China) Investment Co., Ltd of Novi) behind the stirring hydrolysis 1h, add again the aspartic protease be equivalent to protein quality 0.5% in the zein slurry (outstanding can the food-grade acid protease P rotex26L of section, the Wuxi is outstanding can biotechnology company limited of section) and stir hydrolysis 4h;
(4) go out enzyme, decolouring: enzymolysis is warming up to 80 ℃ after finishing, add 0.2% activated carbon powder (400 order) stirring and evenly mixing be equivalent to protein wt in the zein slurries, constant temperature 36rpm stirs 30min, contains L-Ala and leucic oligopeptide content in the enzyme of realizing going out, adsorption bleaching and the lifting product;
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slush pump that will contain activated carbon enters plate and frame diatomite filter coarse filtration, clear liquid after the press filtration at first is the ultra-filtration membrane ultrafiltration of 100kDa by molecular weight cut-off, filtered solution passes through the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa again, filtered solution is the nanofiltration membrane nanofiltration of 1nm again through the aperture, finish desalination, remove total free aminoacids and concentrated, concentrated solution drying tower is dry, inlet temperature is controlled to be 160 ℃, temperature out is controlled to be 70 ℃, the dry end, packing namely obtains the high weak bitter taste maize oligopeptide of L-Ala and leucine content.
Embodiment 3
(1) dispersion of Zein powder, aquation: with Zein powder (protein content 〉=60%, be purchased from Shandong west Wang Jituan) be crushed to 80 orders, with deionized water Zein powder being mixed with mass percent is 5% corn protein solution, with stirrer with the rotating speed of 2500rpm with Zein powder disperse, aquation 15min;
(2) gelatinization, degreasing, destarching: be 1mol.L with volumetric molar concentration
-1Sodium hydroxide solution the Zein powder pH value of solution is adjusted to 5.0, be heated to 85 ℃, and keep 20min so that the zein molecular structure fully launches, and make wherein the abundant gelatinization of starch; Add 0.1% α-alpha-amylase (the SUKAMy-Hi LQ20 be equivalent to the Zein powder quality, Su Kehan biotechnology company limited), enzymolysis 30min under 36rpm, 85 ℃ of conditions, after enzymolysis finished, enzymolysis solution kept 10min, the activity of α-passivation alpha-amylase in 100 ℃, then cooling down to 55 ℃, adopt separating centrifuge with the centrifugal 20min of 4000r/min rotating speed, the purification Zein powder after degrease and liquid, solid slag are degreasing and remove starch;
(3) enzymolysis: with 50 ℃ of deionized waters the solid slag dispersed with stirring is become the zein slurry, guarantee that protein content is 5% in the slurries; Regulate slurry pH value to 6.0, under 50 ℃ of constant temperature, add the neutral protease (Neutrase0.8L that is equivalent to protein quality 0.5% in the zein slurry, letter (China) Investment Co., Ltd of Novi) behind the stirring hydrolysis 2h, add again the aspartic protease be equivalent to protein quality 2% in the zein slurry (outstanding can the food-grade acid protease P rotex26L of section, the Wuxi is outstanding can biotechnology company limited of section) and stir hydrolysis 8h;
(4) go out enzyme, decolouring: enzymolysis is warming up to 85 ℃ after finishing, add 1% activated carbon powder (200 order) stirring and evenly mixing be equivalent to protein wt in the zein slurries, constant temperature 36rpm stirs 60min, contains L-Ala and leucic oligopeptide content in the enzyme of realizing going out, adsorption bleaching and the lifting product;
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slush pump that will contain activated carbon enters plate and frame diatomite filter coarse filtration, clear liquid after the press filtration at first is the ultra-filtration membrane ultrafiltration of 100kDa by molecular weight cut-off, filtered solution passes through the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa again, filtered solution is the nanofiltration membrane nanofiltration of 1nm again through the aperture, finish desalination, remove total free aminoacids and concentrated, concentrated solution drying tower is dry, inlet temperature is controlled to be 165 ℃, temperature out is controlled to be 90 ℃, the dry end, packing namely obtains the high weak bitter taste maize oligopeptide of L-Ala and leucine content.
Embodiment 4
(1) dispersion of Zein powder, aquation: with Zein powder (protein content 〉=60%, be purchased from Shandong west Wang Jituan) be crushed to 80 orders, with deionized water Zein powder being mixed with mass percent is 15% corn protein solution, with stirrer with the rotating speed of 3000rpm with Zein powder disperse, aquation 15min;
(2) gelatinization, degreasing, destarching: be 4mol.L with volumetric molar concentration
-1Sodium hydroxide solution the Zein powder pH value of solution is adjusted to 6.0, be heated to 80 ℃, and keep 30min so that the zein molecular structure fully launches, and make wherein the abundant gelatinization of starch; Add 0.8% α-alpha-amylase (the SUKAMy-Hi LQ20 be equivalent to the Zein powder quality, Su Kehan biotechnology company limited), enzymolysis 50min under 36rpm, 80 ℃ of conditions, after enzymolysis finished, enzymolysis solution kept 20min, the activity of α-passivation alpha-amylase in 95 ℃, then cooling down to 60 ℃, adopt separating centrifuge with the centrifugal 20min of 5000r/min rotating speed, the purification Zein powder after degrease and liquid, solid slag are degreasing and remove starch;
(3) enzymolysis: with 55 ℃ of deionized waters the solid slag dispersed with stirring is become the zein slurry, guarantee that protein content is 10% in the slurries; Regulate slurry pH value to 6.5, under 55 ℃ of constant temperature, add the neutral protease (Neutrase0.8L that is equivalent to protein quality 0.5% in the zein slurry, letter (China) Investment Co., Ltd of Novi) behind the stirring hydrolysis 1.5h, add again the aspartic protease be equivalent to protein quality 0.5% in the zein slurry (outstanding can the food-grade acid protease P rotex26L of section, the Wuxi is outstanding can biotechnology company limited of section) and stir hydrolysis 6h;
(4) go out enzyme, decolouring: enzymolysis is warming up to 80 ℃ after finishing, add 0.4% activated carbon powder (200 order) stirring and evenly mixing be equivalent to protein wt in the zein slurries, constant temperature 36rpm stirs 40min, contains L-Ala and leucic oligopeptide content in the enzyme of realizing going out, adsorption bleaching and the lifting product;
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slush pump that will contain activated carbon enters plate and frame diatomite filter coarse filtration, clear liquid after the press filtration at first is the ultra-filtration membrane ultrafiltration of 100kDa by molecular weight cut-off, filtered solution passes through the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa again, filtered solution is the nanofiltration membrane nanofiltration of 1nm again through the aperture, finish desalination, remove total free aminoacids and concentrated, concentrated solution drying tower is dry, inlet temperature is controlled to be 160 ℃, temperature out is controlled to be 70 ℃, the dry end, packing namely obtains the high weak bitter taste maize oligopeptide of L-Ala and leucine content.
Embodiment 5
(1) dispersion of Zein powder, aquation: with Zein powder (protein content 〉=60%, be purchased from Shandong west Wang Jituan) be crushed to 80 orders, with deionized water Zein powder being mixed with mass percent is 15% corn protein solution, with stirrer with the rotating speed of 3000rpm with Zein powder disperse, aquation 15min;
(2) gelatinization, degreasing, destarching: be 4mol.L with volumetric molar concentration
-1Sodium hydroxide solution the Zein powder pH value of solution is adjusted to 6.0, be heated to 80 ℃, and keep 30min so that the zein molecular structure fully launches, and make wherein the abundant gelatinization of starch; Add 0.8% α-alpha-amylase (the SUKAMy-Hi LQ20 be equivalent to the Zein powder quality, Su Kehan biotechnology company limited), enzymolysis 50min under 36rpm, 80 ℃ of conditions, after enzymolysis finished, enzymolysis solution kept 20min, the activity of α-passivation alpha-amylase in 95 ℃, then cooling down to 60 ℃, adopt separating centrifuge with the centrifugal 20min of 5000r/min rotating speed, the purification Zein powder after degrease and liquid, solid slag are degreasing and remove starch;
(3) enzymolysis: with 55 ℃ of deionized waters the solid slag dispersed with stirring is become the zein slurry, guarantee that protein content is 10% in the slurries; Regulate slurry pH value to 6.5, under 55 ℃ of constant temperature, add the neutral protease (Neutrase0.8L that is equivalent to protein quality 0.5% in the zein slurry, letter (China) Investment Co., Ltd of Novi) behind the stirring hydrolysis 1.5h, add again the aspartic protease be equivalent to protein quality 0.5% in the zein slurry (outstanding can the food-grade acid protease P rotex26L of section, the Wuxi is outstanding can biotechnology company limited of section) and stir hydrolysis 6h;
(4) enzyme that goes out: enzymolysis is warming up to 80 ℃ after finishing, and constant temperature 36rpm stirs the 40min enzyme that goes out.
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slush pump behind the enzyme that will go out enters plate and frame diatomite filter coarse filtration, clear liquid after the press filtration at first passes through the ultra-filtration membrane ultrafiltration of molecular weight cut-off 100kDa, filtered solution passes through the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa again, filtered solution is the nanofiltration membrane nanofiltration of 1nm again through the aperture, finish desalination, remove total free aminoacids and concentrated, concentrated solution drying tower is dry, inlet temperature is controlled to be 160 ℃, temperature out is controlled to be 70 ℃, the dry end, packing namely obtains the high weak bitter taste maize oligopeptide of L-Ala and leucine content.
The maize oligopeptide powder that above embodiment 1～4 makes is a little less than 10 veteran personnel's subjective appreciation bitter tastes, direct water dissolving oral, or add the beverage dissolving orals such as milk, fruit juice, and can be numerous crowds and accept, also easily go out sapid food by simple flavor mixing technological development.Embodiment 5 is not for adding the activated carbon adsorption group, and the maize oligopeptide powder that embodiment 5 makes is stronger through 10 veteran personnel's subjective appreciation bitter tastes, need to or add the bitter taste sequestering agent by allotment and improve local flavor raising acceptability.
Adopt respectively the method for GB/T5009.3, GB/T5009.4, GB/T5009.5 regulation to measure maize oligopeptide powder moisture, ash content, the protein content that embodiment 1～5 makes; The maize oligopeptide powder peptide content that employing makes by the 6.3 methods analyst embodiment 1～5 that stipulate among the GB/T22729-2008; The maize oligopeptide powder relative molecular weight that employing GB/T22729-2008 appendix A high performance gel filtration chromatography analysis embodiment 1～5 makes is less than 1000Da peptide proportion; Adopt Yang Yuexin, Wang Guangya chief editor's " edible analyzing food nutrition components of mirowave handbook (China Light Industry Press, first version in 2002) total free aminoacids that provides, the hydrolysis amino acid analytical procedure detects maize oligopeptide powder Free Amino Acids quality and the total amino acid quality that embodiment 1～5 makes, wherein: the L-Ala quality is that L-Ala content deducts free L-Ala quality income value and accounts for total amino acid content ratio in the maize oligopeptide powder in the maize oligopeptide powder in the oligopeptide, leucine content is that the leucine quality deducts the free leucine content income value and accounts for total amino acid quality ratio in the maize oligopeptide powder in the maize oligopeptide powder in the oligopeptide, the results are shown in Table 1.
The maize oligopeptide powder analyzing and testing result that table 1 embodiment 1～5 makes
Annotate: wherein the maize oligopeptide powder that makes of embodiment 1～5 is designated as respectively 1#, 2#, 3#, 4#, 5#.
By as seen from Table 1, the mass percent of protein is 85.42%～87.63% in the maize oligopeptide that employing the inventive method embodiment 1～4 makes, relative molecular weight is 92.44%～94.21% less than the mass percent of 1000Da peptide, the mass percent of L-Ala is 6.75%～6.94%, and leucic mass percent is 14.12%～14.66%; The mass percent of total free aminoacids is 1.12%～1.36%; Embodiment 5 is not for adding the activated carbon adsorption group, all processing parameters are the same with embodiment 4 except adding activated carbon, maize oligopeptide protein content, free aminoacid content that embodiment 5 makes slightly increase, but L-Ala, leucine content obviously descend in the oligopeptide, L-Ala content is 5.86%, leucine content is 12.97%, and visible activated carbon selective adsorption can obviously improve L-Ala in the maize oligopeptide, leucine content.
Above-described embodiment is the better embodiment of the present invention; but embodiments of the present invention are not restricted to the described embodiments; other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (9)
1. the preparation method of the weak bitter taste maize oligopeptide that a L-Ala and leucine content are high is characterized in that: may further comprise the steps:
(1) dispersion of Zein powder, aquation: Zein powder is pulverized, be formulated as corn protein solution with deionized water, with corn protein solution dispersion, aquation;
(2) gelatinization, degreasing, destarching: the pH value of the corn protein solution that adjusting disperses, aquation is good is 5～7, is heated to 80 ℃～85 ℃, and keeps 20min～60min; Add the α-alpha-amylase that is equivalent to Zein powder quality 0.1%～1%, under 80 ℃～85 ℃ conditions, behind stirring enzymolysis 30min～60min, passivation α-high-temperature starch enzymic activity, cooling down, centrifugal, degrease and liquid are got precipitation;
(3) enzymolysis: the precipitation dispersed with stirring that step (2) is obtained with deionized water is the zein slurry, the mass percent that guarantees protein in the zein slurry is 5%～20%, the pH value of adjusting corn protein milk is 6～7, add the neutral protease that is equivalent to protein quality 0.5%～2% in the zein slurry and stir hydrolysis 1h～2h in 50 ℃～55 ℃, add again the aspartic protease be equivalent to protein quality 0.5%～2% in the zein slurry and stir hydrolysis 4h～8h in 50 ℃～55 ℃, finish to enzymolysis;
(4) go out enzyme, decolouring: after enzymolysis finishes, heat up, add the active carbon powder stirring and evenly mixing, constant temperature stirs 30～60min;
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slurry coarse filtration after the enzyme that will go out, the decolouring, clear liquid after the press filtration is by ultrafiltration, and filtered solution passes through nanofiltration again, finishes desalination, removes total free aminoacids and concentrated, drying obtains the high weak bitter taste maize oligopeptide of L-Ala and leucine content.
2. preparation method according to claim 1, it is characterized in that: the quality percentage of protein is more than 60% in the Zein powder described in the step (1);
Described pulverizing refers to Zein powder is crushed to 80 orders;
The mass percent of the final concentration of Zein powder is 5%～20% in the described corn protein solution;
The velocity of shear of described dispersion, aquation is 2000rpm～3000rpm, and the time is 10 ~ 20min.
3. preparation method according to claim 1 is characterized in that: the pH value described in the step (2) is by using sodium hydroxide solution to regulate;
The stirring velocity of described stirring enzymolysis is 36rpm;
Described passivation α-high-temperature starch enzymic activity refers to enzymolysis solution is kept 10min～20min at 95 ℃～100 ℃;
Described cooling down refers to be cooled to 55 ℃～60 ℃;
Described centrifugal be centrifugal 10min～20min under 4000rpm～6000rpm.
4. preparation method according to claim 3, it is characterized in that: the concentration of described sodium hydroxide solution is 1mol.L
-1～5mol.L
-1
5. preparation method according to claim 1, it is characterized in that: the temperature of the deionized water described in the step (3) is 50 ℃～55 ℃.
6. preparation method according to claim 1, it is characterized in that: the intensification described in the step (4) refers to that temperature rises to 80 ℃～85 ℃;
The add-on of described active carbon powder is equivalent to 0.2%～1% of protein quality in the zein slurry; Used gac is the above gacs of 200 orders.
7. preparation method according to claim 1 is characterized in that: the coarse filtration described in the step (5) refers to the zein stock pump after the enzyme that goes out, the decolouring is entered plate and frame diatomite filter coarse filtration;
Described ultrafiltration refers to the first ultra-filtration membrane ultrafiltration by molecular weight cut-off 100kDa of the stillness of night after the press filtration, and filtered solution is the ultra-filtration membrane ultrafiltration by molecular weight cut-off 5kDa again;
Described nanofiltration refers to that filtered solution is the nanofiltration membrane nanofiltration of 1nm through the aperture;
Described drying is dry by drying tower, and it is 160 ℃～170 ℃ that operating parameters is preferably inlet temperature, and temperature out is 70 ℃～90 ℃.
8. a L-Ala and the high weak bitter taste maize oligopeptide of leucine content, each described preparation method obtains by claim 1～7.
9. the high weak bitter taste maize oligopeptide of L-Ala according to claim 1 and leucine content, it is characterized in that: the mass percent of protein is 85.42%～87.63% in the high weak bitter taste maize oligopeptide of described L-Ala and leucine content, relative molecular weight is 92.44%～94.21% less than the mass percent of 1000Da peptide, the mass percent of L-Ala is 6.75%～6.94%, and leucic mass percent is 14.12%～14.66%; The mass percent of total free aminoacids is 112%～136%.
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| CN105481943A (en) * | 2015-12-15 | 2016-04-13 | 北京天肽生物科技有限公司 | Hangover-alleviating liver-protecting oligopeptide powder |
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