CN103409487B - Method used for extracting maize germ active components - Google Patents
Method used for extracting maize germ active components Download PDFInfo
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- CN103409487B CN103409487B CN201310312484.9A CN201310312484A CN103409487B CN 103409487 B CN103409487 B CN 103409487B CN 201310312484 A CN201310312484 A CN 201310312484A CN 103409487 B CN103409487 B CN 103409487B
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Abstract
The invention belongs to the field of biological extraction technologies, and more specifically relates to a method used for extracting maize germ active components. The method comprises following steps: sub-critical fluid extraction of maize oil, wall breaking of germ cake, extraction of globulin with salt, and extraction of maize germ biological active peptides by enzymatic hydrolysis. The method is capable of fully protecting and taking advantage of the active components of maize germ, and biological active peptides are prepared by full utilization of maize germ proteins, so that effective value increase of the by-products generated in the processes of maize deep processing is realized, and social benefits and economic benefits are substantial.
Description
technical field
The invention belongs to technical field of biological extraction, be specifically related to a kind of extracting method of maize germ activeconstituents.
background technology
Corn is annual gramineae plant, originate in America, quattrocento imports China into, present China various places have cultivation, be the important food crop of China, annual production 1.2 hundred million tons, accounts for more than 20% of world's corn ultimate production, being 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, be mainly used in fodder industry as nutritious maize germ, cheap, added value is not high, therefore be necessary to carry out better deep processing and utilization to it, be 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.
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 the present invention is to provide a kind of extracting method of maize germ activeconstituents, fully effectively extract the activeconstituents in maize germ.
The present invention is by the following technical solutions:
An extracting method for maize germ activeconstituents, comprises the following steps: (1) maize germ puts into subcritical abstraction equipment, passes into CO
2extract, solid-liquid separation after extraction, liquid portion is Semen Maydis oil, and solid part is germ cake; (2) pulverize germ cake, add water formation mixed solution, adds wall breaking enzyme and carry out broken wall; (3) broken wall terminates rear interpolation salt and carries out salt and carry; (4) salt is carried through centrifugal acquisition supernatant liquor and precipitation after end, and supernatant liquor is undertaken desalting and concentrating by nanofiltration membrane, is then drying to obtain maize germ sphaeroprotein; (5) in the precipitation after centrifugal, first add water, then add proteolytic enzyme and carry out enzymolysis, after enzymolysis terminates, feed liquid carries out solid-liquid separation, and clear liquid is crossed ultra-filtration membrane and carried out removal of impurities, is concentrated by nanofiltration membrane, is drying to obtain maize germ biologically active peptides.
Extraction conditions in described step (1) is: temperature 40 ~ 60 DEG C, pressure 1 ~ 50MPa, extraction time 10 ~ 300min, CO
2flow 20 ~ 55L/h.
In described step (2), wall breaking enzyme is one or more in middle temperature amylase, polygalacturonase, cellulase, hemicellulase, and the addition of wall breaking enzyme is that it adds 0.01 ~ 3% of forward slip value liquid total mass, and the add-on of water is 5 ~ 10 times of germ cake quality.
When wall breaking enzyme is middle temperature amylase, polygalacturonase, cellulase, hemicellulase four kinds of compound tenses, its composite mass ratio is 1: 1 ~ 2: 3 ~ 7: 2 ~ 5.
Wall breaking enzyme broken wall condition in described step (2) is: temperature 40 ~ 60 DEG C, pH4 ~ 6.5,3 ~ 7 hours time.
In described step (3), salt is one or both in ammonium sulfate, sodium-chlor, and when being two kinds, the mass ratio of ammonium sulfate and sodium-chlor is 1: 1 ~ 3.
In described step (3) addition of salt be germ cake quality 3 ~ 40%, salt carry the time be 1 ~ 4 hour, salt temperature raising degree is 40 ~ 60 DEG C.
Described step (4) and (5) middle drying temperature be not higher than 60 DEG C or adopt spraying dry.
Proteolytic enzyme is in described step (5): one or more in Sumizyme MP, papoid, trypsinase, stomach en-, neutral protease, bromeline, and proteolytic enzyme addition is that it adds 0.05 ~ 5% of forward slip value liquid total mass.
Enzymatic hydrolysis condition in described step (5) is: temperature 40 ~ 60 DEG C, pH 2 ~ 10, time 3 ~ 8 hours, and the add-on of water is 5 ~ 12 times of precipitation quality.
The present invention can utilize the ultra-filtration membrane of different molecular weight to carry out molecular retention to clear liquid, thus obtains the product in different molecular weight interval.
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 broken wall 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 fully extracted sphaeroprotein in maize germ, 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 zymolysis technique 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 make 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.
The maize germ biologically active peptides that the present invention obtains 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.
embodiment
Embodiment 1:
The extracting method of maize germ activeconstituents, comprise the following steps: (1) maize germ extracts Semen Maydis oil through subcritical abstraction still, extraction conditions is: temperature 60 C, pressure 50MPa, extraction time 200min, flow 55L/h, solid-liquid separation after extraction, liquid portion is Semen Maydis oil, and solid part is germ cake; (2) pulverize germ cake, grinding particle size is 100 orders, and the water adding germ cake quality 5 times forms mixed solution, and the compound wall breaking enzyme adding mixed solution quality 0.01% carries out enzymolysis, and enzymatic hydrolysis condition is: temperature 40 DEG C, pH5.5, time 7 hours; (3) broken wall terminates rear interpolation sodium-chlor and carries out salt method sphaeroprotein, and its condition is: the addition of sodium-chlor be germ cake quality 3%, temperature 60 C, the time carried by salt is 1 hour; (4) salt propose end after through centrifugal acquisition supernatant liquor and precipitation, supernatant liquor is undertaken desalting and concentrating by nanofiltration membrane, and then namely spraying dry obtains maize germ sphaeroprotein; (5) upwards walk centrifugal after precipitation in add its 12 times of quality water formed mixed solution, add the Sumizyme MP enzymolysis of mixed solution quality 5% again, enzymatic hydrolysis condition is: temperature 60 C, pH 9.5, time 3 hours, after enzymolysis terminates, feed liquid carries out solid-liquid separation through whizzer, clear liquid is crossed 100,000 molecular weight films and is carried out removal of impurities, and concentrated by nanofiltration membrane, namely spraying dry obtains maize germ biologically active peptides.
Compound wall breaking enzyme used in the present embodiment is: middle temperature amylase, polygalacturonase, cellulase and hemicellulase, and its quality compound proportion is 1: 1: 3: 2.
Embodiment 2:
The extracting method of maize germ activeconstituents, comprise the following steps: (1) maize germ extracts Semen Maydis oil through subcritical abstraction still, extraction conditions is: temperature 40 DEG C, pressure 30MPa, extraction time 100min, flow 20L/h, solid-liquid separation after extraction, liquid portion is Semen Maydis oil, and solid part is germ cake; (2) pulverize germ cake, grinding particle size is 400 orders, and the water adding germ cake quality 10 times forms mixed solution, and the compound wall breaking enzyme adding mixed solution quality 3% carries out enzymolysis, and enzymatic hydrolysis condition is: temperature 55 DEG C, pH6.5, time 5 hours; (3) broken wall terminates rear interpolation ammonium sulfate and carries out salt method sphaeroprotein, and its condition is: the addition of ammonium sulfate be germ cake quality 40%, temperature 50 C, the time carried by salt is 4 hours; (4) salt propose end after through centrifugal acquisition supernatant liquor and precipitation, supernatant liquor is undertaken desalting and concentrating by nanofiltration membrane, and then namely lyophilize obtains maize germ sphaeroprotein; (5) upwards walk centrifugal after precipitation in add its 8 times of quality water formed mixed solution, add the trypsin digestion of mixed solution quality 0.05% again, enzymatic hydrolysis condition is: temperature 40 DEG C, pH 2, time 8 hours, after enzymolysis terminates, feed liquid carries out solid-liquid separation through whizzer, clear liquid is crossed 100,000 molecular weight films and is carried out removal of impurities, and concentrated by nanofiltration membrane, namely lyophilize obtains maize germ biologically active peptides.
Compound wall breaking enzyme used in the present embodiment is: middle temperature amylase, polygalacturonase, cellulase and hemicellulase, and its quality compound proportion is 1: 2: 7: 5.
Embodiment 3:
The extracting method of maize germ activeconstituents, comprise the following steps: (1) maize germ extracts Semen Maydis oil through subcritical abstraction still, extraction conditions is: temperature 50 C, pressure 10MPa, extraction time 50min, flow 30L/h, solid-liquid separation after extraction, liquid portion is Semen Maydis oil, and solid part is germ cake; (2) pulverize germ cake, grinding particle size is 300 orders, and the water adding germ cake quality 8 times forms mixed solution, and the middle temperature amylase adding mixed solution quality 1% carries out enzymolysis, and enzymatic hydrolysis condition is: temperature 60 C, pH4, time 3 hours; (3) broken wall terminates rear interpolation composite salt (mass ratio 1:1 sodium-chlor and ammonium sulfate) and carries out salt method sphaeroprotein, and its condition is: the addition of composite salt be germ cake quality 20%, temperature 40 DEG C, the time carried by salt is 3 hours; (4) salt propose end after through centrifugal acquisition supernatant liquor and precipitation, supernatant liquor is undertaken desalting and concentrating by nanofiltration membrane, and then 40 DEG C are drying to obtain maize germ sphaeroprotein; (5) upwards walk centrifugal after precipitation in add its 5 times of quality water formed mixed solution, add the neutral protease enzymolysis of mixed solution quality 1% again, enzymatic hydrolysis condition is: temperature 50 C, pH 5, time 5 hours, after enzymolysis terminates, feed liquid carries out solid-liquid separation through whizzer, clear liquid is crossed 100,000 molecular weight films and is carried out removal of impurities, is concentrated by nanofiltration membrane, and 40 DEG C are drying to obtain maize germ biologically active peptides.
Measure the composition of embodiment 1-3 Raw maize germ and product maize germ biologically active peptides, compare, measuring method used is as follows:
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 product maize germ biologically active peptides obtained by table 2
| 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 inventive method carries out deep processing to maize germ, the fat in obtained product maize germ biologically active peptides 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 (7)
1. an extracting method for maize germ activeconstituents, is characterized in that comprising the following steps: (1) maize germ puts into subcritical abstraction equipment, passes into CO
2extract, solid-liquid separation after extraction, liquid portion is Semen Maydis oil, and solid part is germ cake; (2) pulverize germ cake, add water formation mixed solution, adds wall breaking enzyme and carry out broken wall; (3) broken wall terminates rear interpolation salt and carries out salt and carry; (4) salt is carried through centrifugal acquisition supernatant liquor and precipitation after end, and supernatant liquor is undertaken desalting and concentrating by nanofiltration membrane, is then drying to obtain maize germ sphaeroprotein; (5) in the precipitation after centrifugal, first add water, then add proteolytic enzyme and carry out enzymolysis, after enzymolysis terminates, feed liquid carries out solid-liquid separation, and clear liquid is crossed ultra-filtration membrane and carried out removal of impurities, is concentrated by nanofiltration membrane, is drying to obtain maize germ biologically active peptides;
Proteolytic enzyme is in described step (5): one or more in Sumizyme MP, trypsinase, neutral protease, and proteolytic enzyme addition is that it adds 0.05 ~ 5% of forward slip value liquid total mass; Enzymatic hydrolysis condition is: temperature 40 ~ 60 DEG C, pH 2 ~ 10, time 3 ~ 8 hours, and the add-on of water is 5 ~ 12 times of precipitation quality.
2. the extracting method of maize germ activeconstituents 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, CO
2flow 20 ~ 55L/h.
3. the extracting method of maize germ activeconstituents as claimed in claim 1 or 2, it is characterized in that: in described step (2), wall breaking enzyme is one or more in middle temperature amylase, polygalacturonase, cellulase, hemicellulase, the addition of wall breaking enzyme is that it adds 0.01 ~ 3% of forward slip value liquid total mass, and the add-on of water is 5 ~ 10 times of germ cake quality.
4. the extracting method of maize germ activeconstituents as claimed in claim 3, is characterized in that: the wall breaking enzyme broken wall condition in described step (2) is: temperature 40 ~ 60 DEG C, pH4 ~ 6.5,3 ~ 7 hours time.
5. the extracting method of maize germ activeconstituents as claimed in claim 1 or 2, is characterized in that: in described step (3), salt is one or both in ammonium sulfate, sodium-chlor, and when being two kinds, the mass ratio of ammonium sulfate and sodium-chlor is 1: 1 ~ 3.
6. the extracting method of maize germ activeconstituents as claimed in claim 5, is characterized in that: in described step (3) addition of salt be germ cake quality 3 ~ 40%, salt carry the time be 1 ~ 4 hour, salt temperature raising degree is 40 ~ 60 DEG C.
7. the extracting method of maize germ activeconstituents as claimed in claim 1 or 2, is characterized in that: described step (4) and (5) middle drying temperature be not higher than 60 DEG C or adopt spraying dry.
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