CN102994598B - 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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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 weak bitter taste maize oligopeptide that a kind of L-Ala and leucine content are high and preparation method thereof.
Background technology
Corn is one of Three major grain crops in the world, Ye Shi China Traditional Agricultural crop.2009, China's corn yield reached 1.64 hundred million tons, accounts for grain yield 20%, occupies 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 W-Gum process is Zein powder (Corn Gluten Meal, CGM), also claim bloom, Zein powder is generally containing the protein of 60% left and right, and main component is zein (68%), gluten (22%) and sphaeroprotein (1.2%).In corn protein, 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, but the essential amino acids content such as Methionin, tryptophane is lower, the amino acid of this uniqueness forms makes 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 is mainly as the feed resource of livestock and poultry cheapness or naturally discharge with waste liquid.According to incompletely statistics, the Zein powder that discharge naturally with waste liquid every year in the whole nation, more than 80,000 tons, has not only been wasted resource, and has been caused environmental pollution.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 makes it have unique physiological function, by scientific and reasonable biotechnology means, can obtain the bioactive peptide with different physiological roles, 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, there is 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 that L-Ala and the leucine concentration by improving in blood produces stable phosphopyridine nucleotide NAD+, strengthen the activity of ethanol dehydrogenase and acetaldehyde dehydrogenase, promote decomposition and the metabolism of ethanol in body, thereby the concentration that reduces 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 particularly existing with oligopeptide form, leucine content are weighed the sober up effect of corn sobering-up peptide.
Natural corn protein molecular is due to effects such as hydrogen bond, hydrophobic bond, disulfide linkage, and peptide chain is curled in protein molecular inside and is similar to spherically, and close structure and have stronger hydrophobicity, is difficult to by protease hydrolysis; In addition, in maize yellow-powder, also there are starch, grease and the pigment etc. of a great deal of, 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 enzymolysis zein, obtains the small molecules amount maize oligopeptide of high yield.Maize oligopeptide is prepared an outstanding difficult problem and is comprised four aspects: (1) product purity is low, and maize oligopeptide protein content is on the low side, is mainly that Zein powder purification processes can not get position in earlier stage, has 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 mouthfeel bitter taste of corn functional peptides is dense even stimulates, very difficult accepted by the public, applies and is restricted; (4) maize oligopeptide has multiple physiological function, but not for a certain given efficacy research and development, product efficacy is not outstanding.As patent CN101531700A discloses, to take maize yellow-powder (protein content 60% left and right) be raw material, employing buck is processed, except destarching, the impurity such as grease obtain high purity zein material slag, again by 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%, neutral protein enzyme dosage is 5%～8%, total enzyme dosage reaches 10%～16%, water consumption, alkali, 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 patent, decolouring is not gone peculiar smell technique, is not described product special flavour in addition, not specifically for a certain effect design of product yet, 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, the dry corn sobering-up peptide powder that obtains of spraying, the peptide section that in this product, molecular weight is less than 1200Da accounts for 73.19%, L-Ala and leucic content are greater than respectively 6.43% and 12.42%, the corn peptide purity of product is low, local flavor situation is not described, in addition because only removing macromolecular substance by roughing out and ultrafiltration in invention, the product Determination of Free Amino Acids obtaining, ash oontent does not provide data, the actual L-Ala existing with 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, after enzymolysis product centrifugation, supernatant liquor obtains through two-stage ultrafiltering membrane sepn the little peptide that main molecules amount is 200～1000Da, this method is large by enzyme amount, reaches 5%～8%.
Summary of the invention
In order to solve difficult point and the weak point of above-mentioned prior art, primary and foremost purpose of the present invention is to provide the preparation method of a kind of L-Ala and the high weak bitter taste maize oligopeptide of leucine content.This preparation method adopts degreasing, destarch to process, in conjunction with enzymolysis, activated carbon selective adsorption, concentrated, dry L-Ala, the weak bitter taste maize oligopeptide that leucine content is high prepared, with low cost, is applicable 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 obtaining 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 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, comprises the following steps:
(1) dispersion of Zein powder, aquation: Zein powder is pulverized, with deionized water, be formulated as corn protein solution, by 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, zein molecular structure is fully launched, and make the abundant gelatinization of starch wherein; Add the α-alpha-amylase that is equivalent to Zein powder quality 0.1%～1%, under 80 ℃～85 ℃ conditions, stir after enzymolysis 30min～60min, passivation α-high-temperature starch enzymic activity, cooling down, centrifugal, degrease and liquid, get precipitation; Precipitation is the purification zein after degreasing and destarching;
(3) enzymolysis: be zein slurry by the precipitation dispersed with stirring that deionized water obtains step (2), the mass percent that guarantees protein in 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 zein slurry to stir hydrolysis 1h～2h in 50 ℃～55 ℃, add again the aspartic protease that is equivalent to protein quality 0.5%～2% in zein slurry to stir hydrolysis 4h～8h in 50 ℃～55 ℃, finish to enzymolysis;
(4) go out enzyme, decolouring: after enzymolysis finishes, heat up, add active carbon powder stirring and evenly mixing, constant temperature stirs 30～60min, realizes go out enzyme, decolouring, lifting L-Ala and leucic content;
(5) coarse filtration, ultrafiltration, nanofiltration: by the zein slurry coarse filtration after the enzyme that goes out, decolouring, clear liquid after press filtration is by ultrafiltration, and filtered solution passes through nanofiltration again, completes desalination, removes total free aminoacids concentrated, dry, obtain the weak bitter taste maize oligopeptide that L-Ala and leucine content are high.
Zein powder described in step (1) (claiming again bloom), requires at least to reach Zein powder NYT685-2003 regulation for feed, preferably one-level feed Zein powder;
In described Zein powder, the mass percent of protein is more than 60%;
Described pulverizing refers to Zein powder is crushed to 80 orders;
In described corn protein solution, the mass percent of the final concentration of Zein powder is 5%～20%;
Described dispersion, the velocity of shear of aquation are 2000rpm～3000rpm, and the time is 10 ~ 20min.
PH value described in step (2) is by being used 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 to 10min～20min at 95 ℃～100 ℃;
Described cooling down refers to and is cooled to 55 ℃～60 ℃;
Described centrifugal be centrifugal 10min～20min under 4000rpm～6000rpm.
The temperature of the deionized water described in step (3) is 50 ℃～55 ℃.
Intensification described in 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 zein slurry; Gac used is gac more than 200 orders.
Coarse filtration described in step (5) refers to the zein stock pump after the enzyme that goes out, decolouring is entered to plate-frame type diatomaceous earth filter coarse filtration;
Described ultrafiltration refers to that stillness of night after press filtration is first by the ultra-filtration membrane ultrafiltration of molecular weight cut-off 100kDa, and filtered solution is again by the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa;
Described nanofiltration refers to the nanofiltration membrane nanofiltration that filtered solution is 1nm through aperture;
Described being dried is preferably dried 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, by aforesaid method, prepared, in preparation-obtained maize oligopeptide, the mass percent of protein is 85.42%～87.63%, the mass percent that relative molecular weight is less than 1000Da peptide is 92.44%～94.21%, 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 Soboring-up liver-protecting product.
Compared with prior art, tool has the following advantages and beneficial effect in the present invention:
(1) the present invention adopts high temperature gelatinization, α-high-temperature starch enzyme liberating, centrifugal separation process, efficiently remove on the one hand starch, the fat in 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 (mass percent that relative molecular weight is less than 1000Da peptide is 92.44%～94.21%), this method energy consumption is low, easy to operate, be easy to realization of industrialization.The imporosity that zein is opened in research at present generally adopts the techniques such as high temperature high pressure process (more than about 121 ℃ processing 10min), micronizing, ultrasonication, or adopt denaturing agent as S-WAT etc., the former is as patent CN101126103A, CN101411494A etc., and energy consumption is high, high to equipment requirements; The latter is as patent CN1786183A etc., and 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 Zein powder of research removal is at present processed centrifugal high temperature again or low-temperature amylase processing centrifuging process, as patent CN101531700A, CN102172293A etc., this method has been removed grease in alkali lye processing is centrifugal, also lost the soluble proteins in Zein powder, reduced product yield simultaneously; Also after having report to adopt organic solvent extraction grease, process again starch technique, have complex operation, residual safety issue.
(2) in enzymolysis process of the present invention, adopt to regulate initial slurry pH value to 6.0～7.0, neutral protease enzymolysis to cause that zein starches pH value and be down to and below 5.0, add aspartic protease 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%, 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 controls selective adsorption coloring matter by the preferred activated carbon of science and absorbing process and bitter substance reaches decolouring, removes peculiar smell, this activated carbon has containing L-Ala, the low feature 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, is also highly susceptible to industrialization.
It is raw material that the present invention is intended to take Zein powder (bloom), adopt degreasing, destarch is processed, in conjunction with gradient enzymolysis, activated carbon selective adsorption, concentrated, the dry maize oligopeptide of preparing weak bitter taste, this peptide protein matter content is more than 85%, relative molecular weight is less than 1000Da oligopeptide and accounts for more than 90%, free aminoacid content is lower than 1.5%, a little less than bitter taste, feature amino acid (L-Ala, leucine) content is high, the L-Ala containing in 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 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, Soboring-up liver-protecting effect is outstanding, can be used as high-quality functional material and is widely used in varieties of food items, 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
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited to this.
Embodiment 1
(1) dispersion of Zein powder, aquation: by Zein powder (protein content >=60%, be purchased from Shandong Xi Wang group) be crushed to 80 orders, with deionized water, Zein powder being mixed with to mass percent is 10% corn protein solution, with stirrer with the rotating speed of 2000rpm by Zein powder disperse, aquation 20min;
(2) gelatinization, degreasing, destarching: by volumetric molar concentration, be 3mol.L
-1sodium hydroxide solution Zein powder pH value of solution is adjusted to 6.0, be heated to 82 ℃, and keep 40min, zein molecular structure is fully launched, and makes the abundant gelatinization of starch wherein; Add 0.5% α-alpha-amylase (the SUKAMy-Hi LQ20 that is equivalent to Zein powder quality, Su Kehan biotechnology company limited), enzymolysis 45min under 36rpm, 82 ℃ of conditions, after enzymolysis finishes, enzymolysis solution keeps 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, degrease and liquid, solid slag is degreasing and removes the purification Zein powder after starch;
(3) enzymolysis: with 55 ℃ of deionized waters, solid slag dispersed with stirring is become to zein slurry, guarantee that in slurries, protein content is 10%; 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 zein slurry, letter (China) Investment Co., Ltd of Novi) stir after hydrolysis 1.5h, add again the aspartic protease (the food-grade acid protease P rotex26L of Jie Neng section, Wuxi biotechnology company limited of Jie Neng section) that is equivalent to protein quality 1% in zein slurry to stir hydrolysis 6h;
(4) go out enzyme, decolouring: after enzymolysis finishes, be warming up to 85 ℃, add 0.6% activated carbon powder (200 order) stirring and evenly mixing that is equivalent to protein wt in zein slurries, constant temperature 36rpm stirs 45min, realizes the enzyme that goes out, adsorption bleaching and promotes in product containing L-Ala and leucic oligopeptide content;
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slush pump containing activated carbon is entered to plate-frame type diatomaceous earth filter coarse filtration, the ultra-filtration membrane ultrafiltration that first clear liquid after press filtration is 100kDa by molecular weight cut-off, filtered solution is again by the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa, the nanofiltration membrane nanofiltration that filtered solution is 1nm through aperture again, complete desalination, remove total free aminoacids concentrated, concentrated solution drying tower is dry, it is 170 ℃ that inlet temperature is controlled, it is 80 ℃ that temperature out is controlled, dry end, packing, obtain the weak bitter taste maize oligopeptide that L-Ala and leucine content are high.
Embodiment 2
(1) dispersion of Zein powder, aquation: by Zein powder (protein content >=60%, be purchased from Shandong Xi Wang group) be crushed to 80 orders, with deionized water, Zein powder being mixed with to mass percent is 20% corn protein solution, with stirrer with the rotating speed of 3000rpm by Zein powder disperse, aquation 10min;
(2) gelatinization, degreasing, destarching: by volumetric molar concentration, be 5mol.L
-1sodium hydroxide solution Zein powder pH value of solution is adjusted to 7.0, be heated to 80 ℃, and keep 60min, zein molecular structure is fully launched, and makes the abundant gelatinization of starch wherein; Add 1% α-alpha-amylase (the SUKAMy-Hi LQ20 that is equivalent to Zein powder quality, Su Kehan biotechnology company limited), enzymolysis 60min under 36rpm, 80 ℃ of conditions, after enzymolysis finishes, enzymolysis solution keeps 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, degrease and liquid, solid slag is degreasing and removes the purification Zein powder after starch;
(3) enzymolysis: with 50 ℃ of deionized waters, solid slag dispersed with stirring is become to zein slurry, guarantee that in slurries, protein content is 20%; 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 zein slurry, letter (China) Investment Co., Ltd of Novi) stir after hydrolysis 1h, add again the aspartic protease (the food-grade acid protease P rotex26L of Jie Neng section, Wuxi biotechnology company limited of Jie Neng section) that is equivalent to protein quality 0.5% in zein slurry to stir hydrolysis 4h;
(4) go out enzyme, decolouring: after enzymolysis finishes, be warming up to 80 ℃, add 0.2% activated carbon powder (400 order) stirring and evenly mixing that is equivalent to protein wt in zein slurries, constant temperature 36rpm stirs 30min, realizes the enzyme that goes out, adsorption bleaching and promotes in product containing L-Ala and leucic oligopeptide content;
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slush pump containing activated carbon is entered to plate-frame type diatomaceous earth filter coarse filtration, the ultra-filtration membrane ultrafiltration that first clear liquid after press filtration is 100kDa by molecular weight cut-off, filtered solution is again by the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa, the nanofiltration membrane nanofiltration that filtered solution is 1nm through aperture again, complete desalination, remove total free aminoacids concentrated, concentrated solution drying tower is dry, it is 160 ℃ that inlet temperature is controlled, it is 70 ℃ that temperature out is controlled, dry end, packing, obtain the weak bitter taste maize oligopeptide that L-Ala and leucine content are high.
Embodiment 3
(1) dispersion of Zein powder, aquation: by Zein powder (protein content >=60%, be purchased from Shandong Xi Wang group) be crushed to 80 orders, with deionized water, Zein powder being mixed with to mass percent is 5% corn protein solution, with stirrer with the rotating speed of 2500rpm by Zein powder disperse, aquation 15min;
(2) gelatinization, degreasing, destarching: by volumetric molar concentration, be 1mol.L
-1sodium hydroxide solution Zein powder pH value of solution is adjusted to 5.0, be heated to 85 ℃, and keep 20min, zein molecular structure is fully launched, and makes the abundant gelatinization of starch wherein; Add 0.1% α-alpha-amylase (the SUKAMy-Hi LQ20 that is equivalent to Zein powder quality, Su Kehan biotechnology company limited), enzymolysis 30min under 36rpm, 85 ℃ of conditions, after enzymolysis finishes, enzymolysis solution keeps 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, degrease and liquid, solid slag is degreasing and removes the purification Zein powder after starch;
(3) enzymolysis: with 50 ℃ of deionized waters, solid slag dispersed with stirring is become to zein slurry, guarantee that in slurries, protein content is 5%; 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 zein slurry, letter (China) Investment Co., Ltd of Novi) stir after hydrolysis 2h, add again the aspartic protease (the food-grade acid protease P rotex26L of Jie Neng section, Wuxi biotechnology company limited of Jie Neng section) that is equivalent to protein quality 2% in zein slurry to stir hydrolysis 8h;
(4) go out enzyme, decolouring: after enzymolysis finishes, be warming up to 85 ℃, add 1% activated carbon powder (200 order) stirring and evenly mixing that is equivalent to protein wt in zein slurries, constant temperature 36rpm stirs 60min, realizes the enzyme that goes out, adsorption bleaching and promotes in product containing L-Ala and leucic oligopeptide content;
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slush pump containing activated carbon is entered to plate-frame type diatomaceous earth filter coarse filtration, the ultra-filtration membrane ultrafiltration that first clear liquid after press filtration is 100kDa by molecular weight cut-off, filtered solution is again by the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa, the nanofiltration membrane nanofiltration that filtered solution is 1nm through aperture again, complete desalination, remove total free aminoacids concentrated, concentrated solution drying tower is dry, it is 165 ℃ that inlet temperature is controlled, it is 90 ℃ that temperature out is controlled, dry end, packing, obtain the weak bitter taste maize oligopeptide that L-Ala and leucine content are high.
Embodiment 4
(1) dispersion of Zein powder, aquation: by Zein powder (protein content >=60%, be purchased from Shandong Xi Wang group) be crushed to 80 orders, with deionized water, Zein powder being mixed with to mass percent is 15% corn protein solution, with stirrer with the rotating speed of 3000rpm by Zein powder disperse, aquation 15min;
(2) gelatinization, degreasing, destarching: by volumetric molar concentration, be 4mol.L
-1sodium hydroxide solution Zein powder pH value of solution is adjusted to 6.0, be heated to 80 ℃, and keep 30min, zein molecular structure is fully launched, and makes the abundant gelatinization of starch wherein; Add 0.8% α-alpha-amylase (the SUKAMy-Hi LQ20 that is equivalent to Zein powder quality, Su Kehan biotechnology company limited), enzymolysis 50min under 36rpm, 80 ℃ of conditions, after enzymolysis finishes, enzymolysis solution keeps 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, degrease and liquid, solid slag is degreasing and removes the purification Zein powder after starch;
(3) enzymolysis: with 55 ℃ of deionized waters, solid slag dispersed with stirring is become to zein slurry, guarantee that in slurries, protein content is 10%; 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 zein slurry, letter (China) Investment Co., Ltd of Novi) stir after hydrolysis 1.5h, add again the aspartic protease (the food-grade acid protease P rotex26L of Jie Neng section, Wuxi biotechnology company limited of Jie Neng section) that is equivalent to protein quality 0.5% in zein slurry to stir hydrolysis 6h;
(4) go out enzyme, decolouring: after enzymolysis finishes, be warming up to 80 ℃, add 0.4% activated carbon powder (200 order) stirring and evenly mixing that is equivalent to protein wt in zein slurries, constant temperature 36rpm stirs 40min, realizes the enzyme that goes out, adsorption bleaching and promotes in product containing L-Ala and leucic oligopeptide content;
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slush pump containing activated carbon is entered to plate-frame type diatomaceous earth filter coarse filtration, the ultra-filtration membrane ultrafiltration that first clear liquid after press filtration is 100kDa by molecular weight cut-off, filtered solution is again by the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa, the nanofiltration membrane nanofiltration that filtered solution is 1nm through aperture again, complete desalination, remove total free aminoacids concentrated, concentrated solution drying tower is dry, it is 160 ℃ that inlet temperature is controlled, it is 70 ℃ that temperature out is controlled, dry end, packing, obtain the weak bitter taste maize oligopeptide that L-Ala and leucine content are high.
Embodiment 5
(1) dispersion of Zein powder, aquation: by Zein powder (protein content >=60%, be purchased from Shandong Xi Wang group) be crushed to 80 orders, with deionized water, Zein powder being mixed with to mass percent is 15% corn protein solution, with stirrer with the rotating speed of 3000rpm by Zein powder disperse, aquation 15min;
(2) gelatinization, degreasing, destarching: by volumetric molar concentration, be 4mol.L
-1sodium hydroxide solution Zein powder pH value of solution is adjusted to 6.0, be heated to 80 ℃, and keep 30min, zein molecular structure is fully launched, and makes the abundant gelatinization of starch wherein; Add 0.8% α-alpha-amylase (the SUKAMy-Hi LQ20 that is equivalent to Zein powder quality, Su Kehan biotechnology company limited), enzymolysis 50min under 36rpm, 80 ℃ of conditions, after enzymolysis finishes, enzymolysis solution keeps 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, degrease and liquid, solid slag is degreasing and removes the purification Zein powder after starch;
(3) enzymolysis: with 55 ℃ of deionized waters, solid slag dispersed with stirring is become to zein slurry, guarantee that in slurries, protein content is 10%; 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 zein slurry, letter (China) Investment Co., Ltd of Novi) stir after hydrolysis 1.5h, add again the aspartic protease (the food-grade acid protease P rotex26L of Jie Neng section, Wuxi biotechnology company limited of Jie Neng section) that is equivalent to protein quality 0.5% in zein slurry to stir hydrolysis 6h;
(4) enzyme that goes out: be warming up to 80 ℃ after enzymolysis finishes, constant temperature 36rpm stirs the 40min enzyme that goes out.
(5) coarse filtration, ultrafiltration, nanofiltration: the zein slush pump going out after enzyme is entered to plate-frame type diatomaceous earth filter coarse filtration, clear liquid after press filtration is first by the ultra-filtration membrane ultrafiltration of molecular weight cut-off 100kDa, filtered solution is again by the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa, the nanofiltration membrane nanofiltration that filtered solution is 1nm through aperture again, complete desalination, remove total free aminoacids concentrated, concentrated solution drying tower is dry, it is 160 ℃ that inlet temperature is controlled, it is 70 ℃ that temperature out is controlled, dry end, packing, obtain the weak bitter taste maize oligopeptide that L-Ala and leucine content are high.
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 by simple flavor mixing technological development, go out sapid food.Embodiment 5 is for not adding 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 improve acceptability by allocating or adding bitter taste sequestering agent to improve local flavor.
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 methods analyst embodiment 1～5 of 6.3 regulations in GB/T22729-2008, the maize oligopeptide powder relative molecular weight that adopts GB/T22729-2008 appendix A high performance gel filtration chromatography analysis embodiment 1～5 to make is less than 1000Da peptide proportion, adopt Yang Yuexin, Wang Guangya chief editor's the edible analyzing food nutrition components of mirowave handbook > > of < < (China Light Industry Press, first version in 2002) total free aminoacids providing, 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: in oligopeptide, L-Ala quality is that in maize oligopeptide powder, L-Ala content deducts free L-Ala quality income value and accounts for total amino acid content ratio in maize oligopeptide powder, in oligopeptide, leucine content is that in maize oligopeptide powder, leucine quality deducts free leucine content income value and accounts for total amino acid quality ratio in maize oligopeptide powder, the results are shown in Table 1.
The maize oligopeptide powder analyzing and testing result that table 1 embodiment 1～5 makes
Note: the maize oligopeptide powder that wherein embodiment 1～5 makes is designated as respectively 1#, 2#, 3#, 4#, 5#.
From table 1, in the maize oligopeptide that employing the inventive method embodiment 1～4 makes, the mass percent of protein is 85.42%～87.63%, the mass percent that relative molecular weight is less than 1000Da peptide is 92.44%～94.21%, 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 activated carbon adsorption group, except adding activated carbon, all processing parameters are the same with embodiment 4, maize oligopeptide protein content, free aminoacid content that embodiment 5 makes slightly increase, but in oligopeptide, L-Ala, leucine content obviously decline, L-Ala content is 5.86%, leucine content is 12.97%, and visible activated carbon selective adsorption can obviously improve L-Ala in maize oligopeptide, leucine content.
Above-described embodiment is preferably 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 principle, substitutes, combination, simplify; all should be equivalent substitute mode, within being included in protection scope of the present invention.
Claims (8)
1. a preparation method for the weak bitter taste maize oligopeptide that L-Ala and leucine content are high, is characterized in that: comprise the following steps:
(1) dispersion of Zein powder, aquation: Zein powder is pulverized, with deionized water, be formulated as corn protein solution, by 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 20 min～60min; Add the α-alpha-amylase that is equivalent to Zein powder quality 0.1%～1%, under 80 ℃～85 ℃ conditions, stir after enzymolysis 30 min～60min, passivation α-high-temperature starch enzymic activity, cooling down, centrifugal, degrease and liquid, get precipitation;
(3) enzymolysis: be zein slurry by the precipitation dispersed with stirring that deionized water obtains step (2), the mass percent that guarantees protein in 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 zein slurry to stir hydrolysis 1h～2h in 50 ℃～55 ℃, add again the aspartic protease that is equivalent to protein quality 0.5%～2% in zein slurry to stir hydrolysis 4 h～8h in 50 ℃～55 ℃, finish to enzymolysis;
(4) go out enzyme, decolouring: after enzymolysis finishes, heat up, add active carbon powder stirring and evenly mixing, constant temperature stirs 30～60min;
(5) coarse filtration, ultrafiltration, nanofiltration: by the zein slurry coarse filtration after the enzyme that goes out, decolouring, clear liquid after press filtration is by ultrafiltration, and filtered solution passes through nanofiltration again, completes desalination, removes total free aminoacids concentrated, dry, obtain the weak bitter taste maize oligopeptide that L-Ala and leucine content are high;
The add-on of the active carbon powder described in step (4) is equivalent to 0.2%～1% of protein quality in zein slurry; Gac used is gac more than 200 orders;
In the high weak bitter taste maize oligopeptide of the described L-Ala obtaining and leucine content, the mass percent of L-Ala is 6.75%～6.94%, and leucic mass percent is 14.12%～14.66%.
2. preparation method according to claim 1, is characterized in that: in the Zein powder described in step (1), the quality percentage of protein is more than 60%;
Described pulverizing refers to Zein powder is crushed to 80 orders;
In described corn protein solution, the mass percent of the final concentration of Zein powder is 5%～20%;
Described dispersion, the velocity of shear of aquation are 2000rpm～3000rpm, and the time is 10～20min.
3. preparation method according to claim 1, is characterized in that: the pH value described in step (2) is by being used 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 to 10 min～20min at 95 ℃～100 ℃;
Described cooling down refers to and is cooled to 55 ℃～60 ℃;
Described centrifugal be centrifugal 10 min～20min under 4000rpm～6000rpm.
4. preparation method according to claim 3, is characterized in that: the concentration of described sodium hydroxide solution is 1 mol.L
-1～5 mol.L
-1.
5. preparation method according to claim 1, is characterized in that: the temperature of the deionized water described in step (3) is 50 ℃～55 ℃.
6. preparation method according to claim 1, is characterized in that: the intensification described in 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 zein slurry; Gac used is gac more than 200 orders.
7. preparation method according to claim 1, is characterized in that: the coarse filtration described in step (5) refers to the zein stock pump after the enzyme that goes out, decolouring is entered to plate-frame type diatomaceous earth filter coarse filtration;
Described ultrafiltration refers to that the clear liquid after press filtration first passes through the ultra-filtration membrane ultrafiltration of molecular weight cut-off 100kDa, and filtered solution is again by the ultra-filtration membrane ultrafiltration of molecular weight cut-off 5kDa;
Described nanofiltration refers to the nanofiltration membrane nanofiltration that filtered solution is 1nm through aperture;
Described being dried is dried by drying tower, and operating parameters is that inlet temperature is 160 ℃～170 ℃, and temperature out is 70 ℃～90 ℃.
8. the high weak bitter taste maize oligopeptide of L-Ala and leucine content, obtains by the preparation method described in claim 1～7 any one.
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