CN104418944A - Technology for separating multiple bioactive components in maize germ - Google Patents
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- CN104418944A CN104418944A CN201310409858.9A CN201310409858A CN104418944A CN 104418944 A CN104418944 A CN 104418944A CN 201310409858 A CN201310409858 A CN 201310409858A CN 104418944 A CN104418944 A CN 104418944A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
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- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
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- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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Abstract
The invention relates to a technology for separating multiple bioactive components in maize germ. The technology comprises the following steps: 1, putting the maize germ into a subcritical extraction kettle, and extracting maize oil; 2, performing enzymolysis on defatted germ by adopting a composite wall-breaking enzyme, so that the germ tissue structure is damaged; 3, adding a composite salt after the wall-breaking operation is ended, and performing salt extraction on globulin; 4, centrifuging after the salt extraction is ended, desalting and concentrating the supernatant by using a nanofiltration membrane, and drying, thereby obtaining maize germ globulin; and preparing a biological active peptide of the maize germ from a precipitation part by virtue of a composite enzyme; and 5, performing solid-liquid separation on feed liquid by virtue of a centrifugal machine after the enzymolysis is ended, performing membrane filtration and membrane concentration on the liquor, and drying, thereby obtaining the biological active peptide of the maize germ. According to the technology disclosed by the invention, the active ingredients in the maize germ are fully protected and utilized, and the biological active peptide is prepared by fully utilizing the maize germ on the basis. The values of by-products in the corn deep-processing process are effectively added, and the social benefits and economical benefits are very obvious.
Description
Technical field:
The invention belongs to biological technical field, be specifically related to the isolation technique of the multiple bioactive ingredients of maize germ.
Background technology:
Corn is annual gramineae plant, originates in America, and quattrocento imports China into, and present China various places have cultivation.Be the important food crop of China, annual production 1.2 hundred million tons, accounting for more than 20% of world's corn ultimate production, is one of Three major grain crops, is also the food crop that worldwide production is the highest.But China's corn deep processing utilize present situation unsatisfactory.From current corn deep processing present situation, corn resources mainly make use of the W-Gum of 70%, mainly for the production of alcohol, starch and β-amylose, fodder industry is mainly used in as nutritious maize germ, cheap, added value is not high, is therefore necessary to carry out better deep processing and utilization to it, is worth thus the increment realizing product to give full play to it.
Corn kernel by seed coat, pericarp, endosperm and embryo four part form.Maize germ is positioned at the bottom of corn kernel side, is the starting point of corn growth and growth.Though maize germ quality only accounts for 11% ~ 14% of seed, nutritive value is higher than endosperm, it concentrated the protein of 22% in corn kernel, the mineral substance of 83% and 84% fat.The crude fat content about 35% ~ 56% of maize germ, accounts for 85% of corn oil-containing total amount.So maize germ is mainly as a kind of oil resource, for producing Fructus Maydis oil.Corn germ protein is a kind of quality protein, nutritious, also containing functional proteins such as the sphaeroprotein with immunologic function.
Linoleic acid content in Semen Maydis oil, up to 65%, is the highest in cereal, and it acts synergistically with the vitamin-E in maize germ, can reduce serum cholesterol concentration and prevent it to be deposited on vessel wall.Therefore, Semen Maydis oil has certain preventive and therapeutic action to coronary heart disease, atherosclerosis, hyperlipidaemia and hypertension etc.Vitamin-E also can promote that human body cell divides, and delays senility.Carotene contained in Semen Maydis oil, can be converted into vitamin A after being absorbed by the body, it has protective effect on cancer risk; Plant sterol in Semen Maydis oil has anti-oxidant, the physiological action such as anti-inflammatory, decreasing cholesterol.In addition, eat Semen Maydis oil more and cancer therapy drug can also be suppressed the side effect of human body, stimulate brain cell, strengthen the mental and memory of people.
Corn germ protein is formed primarily of white protein, sphaeroprotein, prolamine and alkali-soluble protein, there is research display: albumin content in corn germ protein is up to 41%, next is glutelin content is 19%, and content of prolamine is minimum is only 3%, and wherein sphaeroprotein content is 14%.
Sphaeroprotein has the effect of enhancing body resistibility.Sphaeroprotein can be precipitated by 50% saturation ratio ammoniumsulphate soln, is insoluble to pure water and is dissolved in the protein of weak brine.At present, to animal globulin, particularly the research of human body sphaeroprotein is very deep, and research range rises to clinical application from theoretical investigation.Fewer to phytoglobulin research, mainly concentrate on the extraction of phytoglobulin, detection and application, because humans and animals sphaeroprotein removes virus difficulty, its safety in utilization is reduced, and people more and more pay close attention to the application of phytoglobulin.
Maize germ biologically active peptides is the polypeptide mixture of the certain molecular weight scope that corn germ protein is formed after directed enzymically hydrolyse, there is good solubility, mobility, thermostability, and its viscosity is low, concentration is high, absorb fast, utilization ratio advantages of higher in vivo, oxidation-resistance, Ginseng Extract, reduction blood sugar concentration, hypotensive, the physiologically active that promotes alcohol metabolism (sobering up) etc. unique can also be discharged when it becomes the less dipeptides of molecular weight, tripeptides.Research and utilization corn germ protein, exploitation novelty teabag, improves its comprehensive utilization value, extends intensive processing industry chain (supply chain), has become a current important subject; Also can be the Application and Development of similar biologically active peptides in food, medicine, healthcare products provides new thinking and countermeasure simultaneously.
To sum up, the utilization of maize germ should take into full account the various activeconstituentss in maize germ, utilize advanced biotechnological means, original composition of maximum reservation maize germ, and prepare new bioactive ingredients on this basis, thus reach effective comprehensive utilization of maize germ, at utmost play its biological value, for mankind's health care belt carrys out welfare.There is very large defect in current enterprise in the deep processing of maize germ, only be confined to the extraction of Fructus Maydis oil, and mostly in the process extracting Fructus Maydis oil only focus on quantity and not quality awareness, greatly reduce effective utilization of maize germ bioactive ingredients.Therefore we are necessary to utilize new technology that maize germ bioactive ingredients is retained fully, and prepare new bioactive ingredients on this basis, reach the object making full use of maize germ resource.
Summary of the invention:
The object of this invention is to provide the isolation technique of the multiple bioactive ingredients of a kind of maize germ, the product bioactive ingredients content of gained is many and active high, is easily absorbed by body, can effectively regulating intestinal canal is healthy, and provides certain immunologic function.
For achieving the above object, the present invention is by the following technical solutions:
An isolation technique for the multiple bioactive ingredients of maize germ, comprises the following steps: comprise the following steps: (1) maize germ puts into subcritical abstraction still, passes into CO
2extract, obtain the Semen Maydis oil of biological activity high-content.(2) pulverize the germ cake after extraction, grinding particle size is 100 ~ 400 orders, adopts compound wall breaking enzyme to carry out enzymolysis to it, is conducive to the extraction of sphaeroprotein and the preparation of bioactive peptide to destroy embryonic tissue structure.(3) broken wall terminates rear interpolation composite salt and carries out salt method sphaeroprotein, and its condition is: the addition of composite salt is 3 ~ 40%, salt carry the time be 1 ~ 4 hour, salt temperature raising degree is 40 ~ 60 DEG C.(4) salt propose end after through centrifugal, supernatant liquor is undertaken desalting and concentrating by nanofiltration membrane, is then drying to obtain maize germ sphaeroprotein; Sediment fraction prepares maize germ biologically active peptides by prozyme.(5) after enzymolysis terminates, feed liquid carries out solid-liquid separation through whizzer, and clear liquid is crossed 100,000 molecular weight films and carried out removal of impurities, is concentrated, be drying to obtain maize germ biologically active peptides by nanofiltration membrane.The film of different molecular weight can be utilized to carry out molecular retention to clear liquid, thus obtain the product in different molecular weight interval.
Extraction conditions in described step (1) is: temperature 40 ~ 60 DEG C, pressure 1-50MPa, extraction time 10 ~ 300min, flow 20 ~ 55L/h.
Compound wall breaking enzyme in described step (2) is: one or more in middle temperature amylase, polygalacturonase, cellulase, hemicellulase; Its compound proportion is: 1: 1 ~ 2: 3 ~ 7: 2 ~ 5; The addition of prozyme is that it adds 0.01 ~ 3% of forward slip value liquid gross weight.
Wall breaking enzyme enzymatic hydrolysis condition in described step (2) is: material-water ratio 1: 5 ~ 10, temperature 40 ~ 60 DEG C, pH4 ~ 6.5,3 ~ 7 hours time.
In described step (3), composite salt is one or both in ammonium sulfate, sodium-chlor, if add two kinds, its compound proportion is: 1: 1 ~ 3.
Described step (4) and (5) middle drying temperature be not higher than 60 DEG C or adopt spraying dry.
Belonging in step (4) prozyme be: one or more in Sumizyme MP, papoid, trypsinase, stomach en-, neutral protease, bromeline; Proteolytic enzyme addition is that it adds 0.05 ~ 5% of forward slip value liquid total amount.
Enzymatic hydrolysis condition in described step (4) is: material-water ratio 1: 5 ~ 12, temperature 40 ~ 60 DEG C, pH 2 ~ 10,3 ~ 8 hours time.
The present invention take maize germ as raw material, by in subcritical equipment extraction plumule Semen Maydis oil, this process operates at low temperatures, make the activeconstituents in Semen Maydis oil such as plant sterol, vitamin A, E etc. obtain better protecting, and protect other bioactive ingredients such as the activated protein in maize germ; Carrying out enzymolysis by wall breaking enzyme to plumule makes the plant tissue of maize germ destroy, and is more conducive to extraction and the preparation of other activeconstituentss in maize germ; Utilize salt method to be extracted sphaeroprotein in maize germ fully, maize germ sphaeroprotein has a lot of physiological function, and effectively can improve human immunological competence; The albumen made full use of in maize germ by complex enzyme zymohydrolysis technology prepares biologically active peptides, maize germ biologically active peptides has good physico-chemical property and processing characteristics and important physiological function, as promoted the absorption of mineral substance, anti-oxidant, hypotensive, reducing blood-fat, decreasing cholesterol, the effect such as antibacterial, can be widely used in food and field of medicaments.The present invention adequately protects and make use of the effective active composition in maize germ, and makes full use of corn germ protein on this basis and prepare biologically active peptides.The byproduct achieved in corn deep processing process effectively rises in value, and Social benefit and economic benefit is all very remarkable.
Embodiment
Embodiment 1:
An isolation technique for the multiple bioactive ingredients of maize germ, comprises the following steps: (1) maize germ extracts Semen Maydis oil through subcritical abstraction equipment, and extraction conditions is: temperature 60 C, pressure 50MPa, extraction time 200min, flow 55L/h.(2) pulverize degreasing plumule, grinding particle size is 100 orders, and adopt compound wall breaking enzyme to carry out enzymolysis to it, enzymatic hydrolysis condition is: material-water ratio 1: 5, temperature 40 DEG C, pH5.5,7 hours time.(3) broken wall terminates rear feed liquid and adds sodium-chlor and carry out salt method sphaeroprotein, and its condition is: the addition of sodium-chlor is 3%, to carry the time be 1 hour to salt.(4) salt propose end after through centrifugal, supernatant liquor is undertaken desalting and concentrating by nanofiltration membrane, and then namely drying machine obtains maize germ sphaeroprotein; Sediment fraction is by Sumizyme MP enzyme-squash techniqued maize germ biologically active peptides, and enzymatic hydrolysis condition is: material-water ratio 1: 12, temperature 60 C, pH 9.5, time 3 hours.(5) after enzymolysis terminates, feed liquid carries out solid-liquid separation through whizzer, and clear liquid is crossed 100,000 molecular weight films and carried out removal of impurities, is concentrated, be drying to obtain maize germ biologically active peptides by nanofiltration membrane.The film of different molecular weight can be utilized to carry out molecular retention to clear liquid, thus obtain the product in different molecular weight interval.
In described step (2), compound wall breaking enzyme is: middle temperature amylase, polygalacturonase, cellulase, hemicellulase, and its compound proportion is: 1: 1: 3: 2; The addition of compound wall breaking enzyme is that it adds 0.01% of forward slip value liquid gross weight.
Spraying dry is adopted to carry out drying in described step (4) and (5).
In affiliated step (4), proteolytic enzyme addition is that it adds 5% of forward slip value liquid total amount.
Embodiment 2:
An isolation technique for the multiple bioactive ingredients of maize germ, comprises the following steps: (1) maize germ extracts Semen Maydis oil through subcritical abstraction equipment, and extraction conditions is: temperature 40 DEG C, pressure 30MPa, extraction time 100min, flow 20L/h.(2) pulverize degreasing plumule, grinding particle size is 400 orders, and adopt compound wall breaking enzyme to carry out enzymolysis to it, enzymatic hydrolysis condition is: material-water ratio 1: 10, temperature 55 DEG C, pH6.5,5 hours time.(3) broken wall terminates rear feed liquid and adds ammonium sulfate and carry out salt method sphaeroprotein, and its condition is: the addition of ammonium sulfate is 40%, to carry the time be 4 hours to salt.(4) salt propose end after through centrifugal, supernatant liquor is undertaken desalting and concentrating by nanofiltration membrane, and then namely drying machine obtains maize germ sphaeroprotein; Sediment fraction is by stomach en-enzyme-squash techniqued maize germ biologically active peptides, and enzymatic hydrolysis condition is: material-water ratio 1: 12, temperature 40 DEG C, pH 2,8 hours time.(5) after enzymolysis terminates, feed liquid carries out solid-liquid separation through whizzer, and clear liquid is crossed 100,000 molecular weight films and carried out removal of impurities, is concentrated, be drying to obtain maize germ biologically active peptides by nanofiltration membrane.The film of different molecular weight can be utilized to carry out molecular retention to clear liquid, thus obtain the product in different molecular weight interval.
In described step (2), compound wall breaking enzyme is: middle temperature amylase, polygalacturonase, cellulase, hemicellulase, and its compound proportion is: 1: 2: 7: 5; The addition of compound wall breaking enzyme is that it adds 3% of forward slip value liquid gross weight.
Lyophilize is adopted in described step (4) and (5).
Affiliated step (4) proteolytic enzyme addition is that it adds 0.05% of forward slip value liquid total amount.
Embodiment 3:
An isolation technique for the multiple bioactive ingredients of maize germ, comprises the following steps: temperature 40 ~ 60 DEG C, pressure 1-50MPa, extraction time 10 ~ 300min, flow 20 ~ 55L/h.(1) maize germ extracts Semen Maydis oil through subcritical abstraction equipment, and extraction conditions is: temperature 50 C, pressure 10MPa, extraction time 300min, flow 30L/h.(2) pulverize degreasing plumule, grinding particle size is 200 orders, and adopt compound wall breaking enzyme to carry out enzymolysis to it, enzymatic hydrolysis condition is: material-water ratio 1: 8, temperature 50 C, pH5.5, time 5 hours.(3) broken wall terminates rear feed liquid and adds composite salt and carry out salt method sphaeroprotein, and its condition is: the addition of composite salt is 6%, salt carry the time be 2 hours, salt temperature raising degree is 50 DEG C.(4) salt propose end after through centrifugal, supernatant liquor is undertaken desalting and concentrating by nanofiltration membrane, is then drying to obtain maize germ sphaeroprotein; Sediment fraction prepares maize germ biologically active peptides by prozyme, and enzymatic hydrolysis condition is: material-water ratio 1: 10, temperature 50 C, pH 8, time 5 hours.(5) after enzymolysis terminates, feed liquid carries out solid-liquid separation through whizzer, and clear liquid is crossed 100,000 molecular weight films and carried out removal of impurities, is concentrated, be drying to obtain maize germ biologically active peptides by nanofiltration membrane.The film of different molecular weight can be utilized to carry out molecular retention to clear liquid, thus obtain the product in different molecular weight interval.
Described step (2) compound wall breaking enzyme is: one or more in middle temperature amylase, polygalacturonase, cellulase, hemicellulase; Its compound proportion is: 1: 1.5: 5: 3; The addition of compound wall breaking enzyme is that it adds 1% of forward slip value liquid gross weight.
In described step (3), composite salt is ammonium sulfate, sodium-chlor, and its compound proportion is: 1: 1.
Described step (4) and (5) middle drying temperature are 40 DEG C.
In affiliated step (4), prozyme is: Sumizyme MP, papoid, trypsinase, neutral protease; Proteolytic enzyme addition is that it adds 1% of forward slip value liquid total amount.
The composition measurement of product:
Determining the protein quantity: micro-Kjeldahl, sample and vitriol oil heat altogether, namely itrogenous organic substance decomposes generation ammonia (digestion), the ammonia produced and effect of sulfuric acid generate ammonium sulfate, then make it to decompose through highly basic alkalization and release ammonia, be evaporated in acid solution by steam by ammonia, the degree be neutralized according to this acid solution can calculate the nitrogen content in sample.
Sphaeroprotein assay: be respectively white protein, sphaeroprotein, gluten, prolamine according to the albumen in the solubleness maize germ of protein.Lipase, cellulase, hemicellulase, amylase is utilized to carry out enzymolysis to maize germ, remove non-proteinaceous and obtain corn germ protein, then the principle of salt solution is dissolved according to sphaeroprotein, ammoniumsulphate soln through 40% extracts sphaeroprotein wherein, and drying is weighed and obtained the content of sphaeroprotein in maize germ.
Little peptide molecular weight measures: by gel filteration determining product Small Peptides molecular weight distribution.Dextrane gel due to different exclusion scope has a specific protein molecule weight range, linear between the logarithm of molecular weight and elution volume within the scope of this, therefore be standard with the protein of several known molecular amount, carry out gel chromatography, with the elution volume of often kind of protein, the logarithm of its molecular weight is mapped, drawing standard elution curve.Unknown sample carries out gel chromatography under same condition, and the elution volume used according to it, can obtain the molecular weight of unknown sample from typical curve.
The determination of fat: adopt the lipid content in soxhlet extraction methods mensuration maize germ.Utilize fat can be dissolved in the character of organic solvent, in apparatus,Soxhlet's, sample anhydrous diethyl ether or sherwood oil equal solvent are extracted repeatedly, after extracting the fat in sample, boil off solvent, the material of gained is the mixture of lipid, to the crude fat content in its i.e. derived sample of weighing.
Albumen and lipid content change in table 1 raw material
Each segment molecule volume production product proportion in table 2 maize germ biologically active peptides
| Molecular weight Da | Embodiment 1 | Embodiment 2 | Embodiment 3 |
| 100-1000 | 88% | 90% | 95% |
| 1000-10000 | 8% | 8% | 4% |
| 10000-100000 | 4% | 2% | 1% |
As shown in Table 1, after the technology of the present invention carries out deep processing to maize germ, the fat in maize germ and protein content all obviously reduce, and the extraction yield of fat and albumen is higher; Sphaeroprotein in maize germ obtains sufficient extraction; The biological activity peptide product major part molecular weight of albumen prepared as shown in Table 2 is distributed in below 1000Da.
Claims (8)
1. an isolation technique for the multiple bioactive ingredients of maize germ, comprises the following steps:
1) maize germ puts into subcritical abstraction still, passes into CO2 and extracts, and obtains the Semen Maydis oil of biological activity high-content.
2) pulverize the germ cake after extraction, grinding particle size is 100 ~ 400 orders, adopts compound wall breaking enzyme to carry out enzymolysis to it, is conducive to the extraction of sphaeroprotein and the preparation of bioactive peptide to destroy embryonic tissue structure.
3) broken wall terminates rear interpolation composite salt and carries out salt method sphaeroprotein, and its condition is: the addition of composite salt is 3 ~ 40%, salt carry the time be 1 ~ 4 hour, salt temperature raising degree is 40 ~ 60 DEG C.
4) salt propose end after through centrifugal, supernatant liquor is undertaken desalting and concentrating by nanofiltration membrane, is then drying to obtain maize germ sphaeroprotein; Sediment fraction prepares maize germ biologically active peptides by prozyme.
5) after enzymolysis terminates, feed liquid carries out solid-liquid separation through whizzer, and clear liquid is crossed 100,000 molecular weight films and carried out removal of impurities, is concentrated, be drying to obtain maize germ biologically active peptides by nanofiltration membrane.The film of different molecular weight can be utilized to carry out molecular retention to clear liquid, thus obtain the product in different molecular weight interval.
2. the isolation technique of the multiple bioactive ingredients of maize germ as claimed in claim 1, is characterized in that: the extraction conditions in described step 1) is: temperature 40 ~ 60 DEG C, pressure 1-50MPa, extraction time 10 ~ 300min, flow 20 ~ 55L/h.
3. the isolation technique of the multiple bioactive ingredients of maize germ as claimed in claim 2, is characterized in that: described step 2) in compound wall breaking enzyme be: one or more in middle temperature amylase, polygalacturonase, cellulase, hemicellulase; Its compound proportion is: 1: 1 ~ 2: 3 ~ 7: 2 ~ 5; The addition of prozyme is that it adds 0.01 ~ 3% of forward slip value liquid gross weight.
4. the isolation technique of the multiple bioactive ingredients of maize germ as claimed in claim 1, is characterized in that: described step 2) in wall breaking enzyme enzymatic hydrolysis condition be: material-water ratio 1: 5 ~ 10, temperature 40 ~ 60 DEG C, pH4 ~ 6.5,3 ~ 7 hours time.
5. the isolation technique of the multiple bioactive ingredients of maize germ as claimed in claim 1, is characterized in that: in described step 3), composite salt is one or both in ammonium sulfate, sodium-chlor, if add two kinds, its compound proportion is: 1: 1 ~ 3.
6. the isolation technique of the multiple bioactive ingredients of maize germ as claimed in claim 1, is characterized in that: described step 4) and 5) middle drying temperature is not higher than 60 DEG C or adopt spraying dry.
7. the isolation technique of the multiple bioactive ingredients of maize germ as claimed in claim 1, is characterized in that: in described step 4), prozyme is: one or more in Sumizyme MP, papoid, trypsinase, stomach en-, neutral protease, bromeline; Proteolytic enzyme addition is that it adds 0.05 ~ 5% of forward slip value liquid total amount.
8. the isolation technique of the multiple bioactive ingredients of maize germ as claimed in claim 1, is characterized in that: the enzymatic hydrolysis condition in described step 4) is: material-water ratio 1: 5 ~ 12, temperature 40 ~ 60 DEG C, pH 2 ~ 10,3 ~ 8 hours time.
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| CN108251479A (en) * | 2018-02-08 | 2018-07-06 | 金华市艾力生物科技有限公司 | A kind of preparation method of maize germ active peptide |
| CN108244329A (en) * | 2018-02-26 | 2018-07-06 | 山东师范大学 | A kind of preparation process of quinoa germ protein powder |
| CN108576313A (en) * | 2018-04-12 | 2018-09-28 | 河南牧业经济学院 | A kind of auxiliary prevents health protection tea and preparation method and the detection method of nonalcoholic fatty liver |
| CN109355338A (en) * | 2018-11-22 | 2019-02-19 | 石家庄爱肽生物科技有限公司 | A kind of preparation method for extracting small-molecular peptides from corn |
| CN111378709A (en) * | 2020-03-20 | 2020-07-07 | 广西壮族自治区农业科学院 | Preparation method of corn polypeptide-selenium chelate |
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