CN101250575A - Method for preparing vanadium-containing peptides and products produced thereby - Google Patents
Method for preparing vanadium-containing peptides and products produced thereby Download PDFInfo
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Abstract
The invention discloses a process for preparing peptide which comprises vanadium, which comprises using stewing material which is rich in protein after scouring with acidulous water solution, mixing with vanadate, using soybean peptide as fermentation accelerating agent, grafting fermenting bacterial strain into raw material which is stewed, fermenting for 6-72h under 20-40 DEG C, washing, drying, disintegrating in ultrafine to obtain vanadium-enriched polypeptide powder, removing peptide with large molecular to obtain vanadium-enriched liquid peptide, and chelating through adding vanadate to obtain vanadium-enriched peptide whose peptide concentration is over 60%, and vanadium element concentration is over 100 mg/kg.
Description
Technical field
The present invention relates to and particularly to contain the preparation method of the peptide of organic vanadium for the preparation method of the biological raw material of food and medicine processing usefulness; The invention still further relates to product by this method preparation.
Background technology
Vanadium is one of human essential trace element.No matter be inorganic vanadium, organic vanadium, the bio-transformation vanadium under threshold limit values, as long as can absorb, just can produce medical care effect.
Organism is very low to the specific absorption of inorganic vanadium, and its positively charged ion maximum absorbance is 10%, and the negatively charged ion specific absorption only is 0.01-0.1%.Simultaneously, the inorganic vanadium toxic side effect is big, and the human body therapy significant quantity is 100-1000 a times of animal experiment toxic dose, and therefore present known inorganic vanadium can not be in clinical application.Than inorganic vanadium, organic vanadium biological activity height, toxicity is low.And human body there is not toxicity with the biological attitude vanadium of protein natural fusion.For example sea cucumber is one of food of only a few enrichment vanadium on the earth, and its ash content vanadiumcontent is up to 15%, but never has the people to poison because of beche-de-mer.Therefore, preparing biological attitude vanadium is a kind of approach safely and effectively for clinical application.
A kind of method that rich vanadium soybean, rich vanadium peanut etc. contain vanadium high-protein food material for preparing has been put down in writing in the applicant's No. 200710302099.0 patent applications, has solved the problem of replenishing v element for human body safety.Yet food proteins just can be absorbed by the body after must being decomposed by stomach and enteral plurality of enzymes earlier.Past attempts thinks that amino acid is the main absorpting form after protein decomposes.The present age, experimental study showed, human body exists amino acid absorption and two kinds of separate transporting mechanisms of peptide absorption to the absorption of nitrogen material.Compare with the amino acid transport system, peptide absorbs rapider, and energy consumption is low, the efficient height, and the amino acid composition more tends to balance, and transports uncontested property and inhibition between the various peptide.And peptide molecule can form inner complex with the metal ion that comprises vanadium, has solubility, is beneficial to body and absorbs.Therefore, with v element and small-peptide chelated making nutritious supplementary, be the preferred form that replenishes v element for human body.In the present age, though by the peptide chelate complex (SPC) of the multiple metallic element of this mechanism preparation as the 4th generation Rare Elements Preparations be widely used in animal rearing, do not see that as yet the record that contains the vanadium polypeptide and contain the little peptide product of vanadium is arranged both at home and abroad.
Summary of the invention
Technical problem to be solved by this invention is, opens up above-mentioned technical field, provides a kind of method of the rich vanadium biological products that can be mass-produced, its product organic vanadium content height, edible safety, specific absorption height, biological value height.
The technical scheme that realizes the object of the invention is as follows:
Choices of raw materials: the root, stem, leaf that can select any suitable mankind plant seed that eat, rich in protein or plant for use are as starting material.Soybean meal, peanut meal behind wherein preferred soybean, peanut and the grease of squeezing out more preferably are rich vanadium soybean and the rich vanadium soybean meal after peanut and the degreasing thereof and the rich vanadium peanut meal of 200710302099.0 Chinese patent application method preparation by application number.
After selected starting material being carried out preliminary working such as necessary cleaning, removal of impurities, process by " acidic aqueous solution washing-boiling-interpolation vanadium compound-fermentation-peptide and vanadate chelating " processing step.
The step 1 washing
Selected raw material earlier with the acidic aqueous solution flushing, is used flushing with clean water again.Used acidic aqueous solution uses the edible acid preparation, and suitable potential of hydrogen is pH3.0-7.5.The kind of used sour material can influence the local flavor of the finished product, therefore can select the edible acid material preparation acidic aqueous solution of different varieties as required for use.Wherein, citric acid, oxysuccinic acid, tartrate, acetic acid all are applicable varieties.
The step 2 boiling
Material after the washing is carried out boiling, and suitable boiling temperature is 100-125 ℃, and cooking time is 20-60 minute.The boiling postcooling is standby to room temperature.
As the optimization method of boiling step, add an amount of other food raw materials in the material before boiling or after the boiling, in order to regulate the local flavor of the finished product.
Step 3 adds vanadium
Treat fermentation materials and spray the vanadium compound aqueous solution.
The step 4 fermentation
(1) uses fermentation accelerant: soybean peptides is dissolved in water, is sprayed on and treats to make fermentation accelerant on the fermentation materials.
(2) inoculation fermentation bacterial strain: the zymophyte that fermentation is used can be any fermented bacterium that is suitable for proteolysis, wherein preferred bacillus CAU208.Fermentation strain is inoculated in treats in the fermentation materials, in 20-40 ℃ of temperature, cultivated 6-72 hour, stop fermentation, sterilization.
(3) preparation contains the peptide class intermediates of v element: after material stops fermentation, clean, drying is pulverized, the rough vanadium polypeptide powder that contains; The rough vanadium polypeptide powder that contains is added the water mill slurry, and slagging-off separates and removes not protolysate, gets liquid rich vanadium polypeptide; The rich vanadium polypeptide of liquid state is concentrated, and the macromole peptide is removed in ultrafiltration, gets the little peptide of liquid rich vanadium; The rich vanadium polypeptide of liquid state is concentrated, and drying gets solid-state rich vanadium polypeptide; The little peptide of the rich vanadium of liquid state is concentrated, and drying gets the little peptide of solid-state rich vanadium.
Above-mentioned processing step two to five fermentation process that embodied are solid state fermentation; The present invention can also use liquid fermentation method, and its concrete processing step is, carries out boiling after the material after the washing is broken into slurries with pigment, directly add vanadium compound afterwards, stirring makes its dissolving, soybean peptides is added the inoculation fermentation bacterial strain again, in 20-40 ℃ of temperature, cultivated 6-72 hour, stop fermentation, sterilization, drying, pulverize, the rough vanadium polypeptide powder that contains.Liquid material defibrination after maybe will fermenting, slagging-off, not protolysate is removed in separation, gets liquid rich vanadium polypeptide, and further work concentrates, drying treatment, gets solid-state rich vanadium polypeptide.
The step 5 chelating
Any intermediates through step 4 fermentation gained, comprise liquid rich vanadium polypeptide, solid-state rich vanadium polypeptide, the little peptide of liquid rich vanadium, the little peptide of solid-state rich vanadium and the rough vanadium polypeptide powder that contains, all can with vanadate aqueous solution chelating, make the higher rich vanadium peptides products of vanadiumcontent.
Its concrete processing method is, contains the peptide intermediates with vanadate solution with the liquid state of step 4 gained and mixes, and being adjusted to vanadium peptide mass ratio is 1: the aqueous solution of 2-4, and heating, constant temperature stirs, and can get the higher rich vanadium peptides products of liquid vanadiumcontent; Further make concentrated, drying treatment, can get the higher rich vanadium peptides products of solid-state vanadiumcontent, comprise the higher liquid rich vanadium polypeptide of vanadiumcontent, solid-state rich vanadium polypeptide, the little peptide of liquid rich vanadium, the little peptide of solid-state rich vanadium.To the solid-state intermediates of step 4 gained, then add the water mill slurry earlier, deal with again.
By method for preparing of the present invention contain the vanadium peptide product, calculate peptide content 〉=60% wherein, v element content 〉=100mg/kg with dry weight.Can be used for making nutritive food, foodstuff additive, nutritious supplementary, medicine etc.
As the optimization method of above-mentioned zymotechnique, have the characteristic of good vanadium enrichment ability according to unicellular lower eukaryote, to fermented bacterium mutagenic and breeding in advance before inoculation, in mutagenic processes, use simultaneously to contain the vanadium substratum.The mutagenesis method of cultivation is ultraviolet mutagenesis or nitrosoguanidine mutagenesis, or carries out ultraviolet ray and the cultivation of nitrosoguanidine complex mutation simultaneously.This method can make the rich vanadium fermented bacterium of high vigor, to improve the product vanadiumcontent and to contain the peptide amount, shortens fermentation period, reduces production costs.Its mutagenesis cultural method is as follows:
(1) preparation contains the vanadium substratum: add an amount of vanadium compound and prepare four kinds of substratum such as activating substratum, seed culture medium, isolation medium, fermention medium respectively, use at different production links.
1. activate substratum:, be used to activate vanadium resistance characteristics by culturing micro-organisms with peptone, glucose, yeast extract paste, potassium primary phosphate, agar, vanadium compound, soybean peptides and water preparation.
2. seed culture medium:, be used to tame vanadium enrichment characteristic by culturing micro-organisms with peptone, glucose, yeast extract paste, potassium primary phosphate, vanadium compound, bean cake powder, soybean peptides and water preparation.
3. isolation medium:, be used for from separated the bacterial strain that using value is arranged by culturing micro-organisms with peptone, glucose, yeast extract paste, potassium primary phosphate, skim-milk, agar, vanadium compound, soybean peptides and water preparation.
4. fermention medium:, be used to measure the proteolysis power and the vanadium enrichment ability of bacterial strain with bean cake powder, peptone, molasses, vanadium compound, soybean peptides and water preparation.
(2) fermented bacterium mutafacient system:
Ultraviolet mutagenesis: bacterial classification is regulated unicellular bacteria suspension viable count to 10 after activating the cultivation of substratum and seed culture medium
6-10
8Individual/ml, liquid layer thickness 0.2-0.4cm is with UV-lamp irradiation mutagenesis.
Nitrosoguanidine mutagenesis: bacterial classification was cultivated about 36 hours through activating substratum and seed culture medium, and regulating viable count is 10
5-10
8Individual/ml, the whole mass concentration of nitrosoguanidine is 0.8-1.2mg/ml, 30 ℃ of water bath processing 30min.
Ultraviolet ray and nitrosoguanidine complex mutation: bacterial classification was cultivated about 36 hours through activating substratum and seed culture medium, and regulating viable count is 10
7-10
9Individual/ml, the whole mass concentration of nitrosoguanidine is 0.8-1.2mg/ml, and 30 ℃ of water bath processing 30min get the liquid layer thickness 0.2-0.4cm after nitrosoguanidine is handled, and the UV-lamp of going is again shone mutagenesis.
(3) strain screening
Primary dcreening operation: primary dcreening operation is just being become bacterial strain behind the ultraviolet mutagenesis; Primary dcreening operation is just being become bacterial strain behind the nitrosoguanidine mutagenesis.
Multiple sieve: the just change bacterial strain that primary dcreening operation is obtained, cultivate 36-60h in the triangular flask fermention medium of access 100ml/250ml, measure the degree of hydrolysis of protein hydrolyzate, multiple sieve, just change bacterial strain that must be more stable, after cultivating in the isolation medium, the transparent circle diameter in the mensuration isolation medium and the ratio of colony diameter, filter out the big bacterial strain of ratio, cultivate through seed culture medium, be inoculated in the fermention medium, the fermented liquid degree of hydrolysis is measured in certain fermentation period (36-60h) back, just change bacterial strain that must be more stable, be divided into two groups of 2 multiple sieves, one group is carried out UV treatment, one group is carried out nitrosoguanidine and handles, select wherein functional and stable bacterial strain and carry out ultraviolet ray and nitrosoguanidine mutagenesis simultaneously, get the superior strain of excellent in stability, superior strain is inoculated into activates on the substratum, went down to posterity 1 time every 5 days later on, passed for 10 generations altogether as the 1st generation, with each for shake flask fermentation (the bottled liquid 100ml of 250ml triangle), measure the degree of hydrolysis of fermented liquid, selected good stability, high yield is good, the regressive bacterial strain of not undergoing mutation is for keeping bacterial strain, the rich vanadium fermentation strain of promptly high vigor.
The rich vanadium fermentation strain of the high vigor heritability of preparation is stable as stated above, v element content can reach 〉=200mg/kg (dry weight), improves 5-8% than maternal bacterial strain degree of hydrolysis, and hydrolysis power improves 35-45%, shorten fermentation period 10-12 hour, production cost reduces 15-25%.
The rich vanadium fermentation strain of the high vigor heritability of preparation is stable as stated above, v element content can reach 〉=200mg/kg (dry weight), improves 5-8% than maternal bacterial strain degree of hydrolysis, and hydrolysis power improves 35-45%, shorten fermentation period 10-12 hour, production cost reduces 15-25%.
The technical process of above-mentioned zymophyte mutagenic and breeding, time parameter, temperature parameter etc. can be adjusted according to different fermented bacteriums.
Above-mentioned zymophyte mutagenesis cultivating method is also replaceable to be protoplastis fusion method or gene engineering research, and the arbitrary combination of also available mutagenesis, protoplastis fusion method, three kinds of methods of gene engineering research is cultivated and used.
The present invention adopts the microbial method fermentation to produce peptide, produces the peptide technology with the chemical method that with the soybean is representative at present with enzymolysis process and compares, and is easy to suitability for industrialized production, saves the energy and water.Particularly adopt the substratum contain vanadium to add ultraviolet ray and the nitrosoguanidine mutagenesis technology is cultivated high vigor richness vanadium fermented bacterium, improve the content of vanadium of fermentation materials, improve hydrolysis rate of protein, add with vanadium compound and peptide huge legendary turtle and close, further improve the v element content of peptide.The rich vanadium polypeptide of gained can be used the ultra-filtration membrane ultrafiltration purification, gets the little peptide of the rich vanadium of high purity, can be used for developing peptide vanadium medicine.V element wherein can be a carrier with little peptide, relies on the absorption system of little peptide uniqueness to be absorbed by body, and its biological value is 2～3 times of inorganic salts additive.
Special when being raw material with the rich vanadium dregs of beans after rich vanadium soybean or the degreasing, the complementary synergy of the biological activity of v element wherein and the biological activity of soybean peptides is specially adapted to prevention or alleviates diabetic complication.Simultaneously, soybean protein is resolved into polypeptide, cut off two kinds of hydrophobic amino acids of hydrophobicity polypeptide end specifically, eliminated the bitter taste that enzymolysis process prepares the beans peptide because in fermentation process, produced abundant proteolytic enzyme.Simultaneously, soybean has been eliminated harmful compositions such as trypsin inhibitor in the soybean, the flatulence factor behind microbial fermentation.
Embodiment
The present invention preparation contains the starting material that method is got of vanadium peptide, and the root, stem, leaf that can select any suitable mankind plant seed that eat, rich in protein or plant for use are as starting material.Soybean meal, peanut meal behind wherein preferred soybean, peanut and the grease of squeezing out more preferably are rich vanadium soybean and the rich vanadium soybean meal after rich vanadium peanut and the degreasing thereof and the rich vanadium peanut meal of 200710302099.0 Chinese patent application method preparation by application number.
200710302099.0 number Chinese patent application prepares the technical essential of rich vanadium edible plant seed, plant seed is done " first vanadium compound, plant growth promoter, seed soak cultivate preservative solution soaking-permanent oxygen of the fixed temperature and humidity of guaranteeing the quality for the first time urge educate-secondary vanadium compound, plant growth promoter, seed soak cultivate preservative solution soaking-secondary guarantee the quality the permanent oxygen of fixed temperature and humidity urge educate-clean, drying ", get to urge for 2 times and educate rich vanadium plant seed.Below in conjunction with embodiment the technology of production vanadium peptide provided by the present invention is done further narration.
Embodiment 1 prepares rich vanadium soybean by No. 200710302099.0 Chinese patent application methods
(1) adds 100 liters in water with 450mg sodium orthovanadate, 10g Plant hormones regulators,gibberellins, 10g potassium bromate, prepare first soak solution.Soybean is cleaned, in the first soak solution of ratio input in 1: 3 (W/V), regulated constant temperature between 16-37 ℃, soak after 8-15 hour, soybean is separated with soak solution, get first soaking soybean.The soak solution of separated and collected can be retained in addition and use.
(2) first soaking soybean is evenly sprayed the seed preservative 0.1% potassium bromate aqueous solution, place 16-37 ℃ of temperature, relative humidity 60-95%, 300-1000M
3/ (th) the permanent oxygen of air output is urged educating to urge in the device and is educated 8-10 hour, and get to urge for the first time and educate soybean.
(3) in the surplus liquid of first immersion, add 150g sodium orthovanadate, 5g Plant hormones regulators,gibberellins, 5g potassium bromate, add water to 100 liters, preparation secondary soak solution.To urge for the first time and educating in the soybean input secondary soak solution, 16-38 ℃ of constant temperature soaks to be cultivated 1-3 hour, and soak solution is separated, and got the secondary soaking soybean.
(4) the secondary soaking soybean is evenly sprayed the nutrient solution of guaranteeing the quality by the 50-100ml/kg consumption, placing temperature is that 16-38 ℃, relative humidity are that 55-95%, air inerchange amount are 300-1000M
3Urge under/(th) the permanent oxygen and educate 1.5-8.5 hour, secondary is urged and is educated soybean.
(5) secondary is urged educate soybean with 1-7% aqueous citric acid solution and flushing with clean water, lyophilize gets the complete rich vanadium soybean of outward appearance, must plant the rich vanadium soybean of skin explosion with other desiccating methods.
The method that present embodiment is used for soyabean processing also can be used for various plants seeds such as peanut, red bean, gets rich accordingly vanadium plant seed material.
Embodiment 2 mutagenesiss are cultivated the rich vanadium fermented bacterium of high vigor
1. bacterial classification is selected
Select any be suitable for proteolysis and edible microorganism for use, for example bacillus, zhizopchin bacterium, graceful radiation mucormycosis, monascus ruber, black-koji mould, graceful radiation mucormycosis, Bacillus subtilus, Bacillus natto, actinomycetes, bacillus pumilus, Aspergillus usamii bacterium variant, cinnamon aspergillus tubigensis, aspergillus oryzae, cereuisiae fermentum, bulgaricus ccm etc.Wherein preferred bacillus CAU208.
2. culture medium preparation
Add vanadium compound and prepare four kinds of substratum such as activation substratum, seed culture medium, isolation medium, fermention medium respectively in different production link uses.
1. activate substratum: be used to activate vanadium resistance characteristics by culturing micro-organisms, its materials and proportioning are, peptone 0.25-0.75%, glucose 0.25-0.75%, yeast extract paste 0.25-0.75%, potassium primary phosphate 0.25-0.75%, agar 1-1.5%, vanadium compound 0.002-0.006%, soybean peptides 1-6% add water to 1000ml.
2. seed culture medium: be used to tame vanadium enrichment characteristic by culturing micro-organisms, its materials and proportioning are, peptone 0.25-0.75%, glucose 0.25-0.75%, yeast extract paste 0.25-0.75%, potassium primary phosphate 0.25-0.75%, bean cake powder 0.1-0.2%, vanadium compound 0.002-0.006%, soybean peptides 1-6% add water to 1000ml.
3. isolation medium: be used for from separated the bacterial strain that using value is arranged by culturing micro-organisms, its materials and proportioning are, peptone 0.25-0.75%, glucose 0.25-0.75%, yeast extract paste 0.25-0.75%, potassium primary phosphate 0.25-0.75%, skim-milk 0.3-1.0%, vanadium compound 0.002-0.006%, soybean peptides 1-6% add water to 1000ml.
4. fermention medium: be used to measure fermentative activity and vanadium enrichment ability that the using value bacterial strain is arranged, its materials and proportioning are, bean cake powder 10%, peptone 0.25-0.75%, molasses 0.77%, vanadium compound 0.002-0.006%, soybean peptides 1-6% add water to 1000ml.
The system component of each substratum and usage ratio can be adjusted according to the mutagenic and breeding of different fermentations bacterial classification.
3. mutafacient system
The ultraviolet mutagenesis method can be used, also the nitrosoguanidine mutagenesis method can be used, and preferred ultraviolet ray and nitrosoguanidine complex mutation method.
(1) ultraviolet mutagenesis: bacterial classification is cultivated about 36h through activating substratum and seed culture medium, regulates unicellular bacteria suspension viable count to 10
6-10
8Individual/ml, liquid layer thickness 0.2-0.4cm, use the 15W-30W UV-lamp at 20cm-30cm apart from irradiation 0.5-4min.
(2) nitrosoguanidine mutagenesis: bacterial classification is cultivated 36h through activating substratum and seed culture medium, and regulating the bacteria suspension viable count is 10
5-10
8Individual/ml, the nitroso guanidine solution for preparing is added in the substratum, the whole mass concentration of nitrosoguanidine is 0.8-1.2mg/ml, 25-35 ℃ of water bath processing 25-35min.
(3) ultraviolet ray and nitrosoguanidine complex mutation: bacterial classification is cultivated 36h through activating substratum and seed culture medium, and regulating the bacteria suspension viable count is 10
7-10
9Individual/ml, the whole mass concentration of nitrosoguanidine is 0.8-1.2mg/ml, 25-35 ℃ of water bath processing 25-35min, and the liquid layer thickness after nitrosoguanidine is handled is 0.2-0.4cm, carry out ultraviolet mutagenesis, use the 15W-30W UV-lamp at 20cm-30cm apart from irradiation 0.5-4min.
(4) strain screening
Primary dcreening operation: behind the ultraviolet mutagenesis, primary dcreening operation is just being become bacterial strain; Behind the nitrosoguanidine mutagenesis, primary dcreening operation is just being become bacterial strain.
Multiple sieve: the just change bacterial strain that primary dcreening operation is obtained, cultivate 36-60h in the triangular flask fermention medium of access 100ml/250ml, measure the degree of hydrolysis of protein hydrolyzate, multiple sieve, obtain more stable just change bacterial strain, put in the isolation medium cultivate 36-60h after, the transparent circle diameter in the mensuration isolation medium and the ratio of colony diameter, filter out the big bacterial strain of ratio, after the seed culture medium bacterium is cultivated, be inoculated in the fermention medium, (36-60h) measures the fermented liquid degree of hydrolysis behind certain fermentation period, just change bacterial strain that must be more stable, be divided into two groups of 2 multiple sieves, one group is carried out ultraviolet mutagenesis, one group is carried out nitrosoguanidine mutagenesis, get functional and stable bacterial strain, again this bacterial strain is carried out ultraviolet ray and nitrosoguanidine mutagenesis simultaneously, get the superior strain of excellent in stability, superior strain is inoculated into activates on the substratum as the 1st generation, went down to posterity 1 time every 5 days later on, passed for 10 generations altogether, each for shake flask fermentation (the bottled liquid 100ml of 250ml triangle), is measured the degree of hydrolysis of fermented liquid, selected good stability, high yield is good, the regressive bacterial strain of not undergoing mutation is the rich vanadium fermented bacterium of high vigor for keeping bacterial strain.
The bean powder of organic vanadium and soybean peptides is rich in the preparation of embodiment 3 solid state fermentations
Make raw material with common soybeans, or make raw material with rich vanadium soybean by embodiment 1 method preparation, pH with 1-7% citric acid or the preparation of other edible acids is the acidic aqueous solution washing of 3.0-7.5 earlier, use flushing with clean water then, drain, every kilogram is sprayed 10-50% vanadium compound aqueous solution 50-500ml, stirs.100-125 ℃ was steamed 30-60 minute, be cooled to room temperature, spray soybean peptides, will be inoculated in the cooled material of boiling by the 1-10% inoculum size by the efficient rich vanadium fermentation strain that embodiment 2 method seed selections are cultivated, in 20-40 ℃ of temperature, cultivate, press air output 1000-3000M
3/ (th) ventilation, per 16 hours stirrings once stopped fermentation after beans is soft through 10-72 hour.Clean beans with 2% citric acid water, 60 ℃ of temperature oven dry, pulverize, gained powder v element content 〉=100mg/kg, water-soluble protein in its nitrogen material 〉=70%, polypeptide 〉=55% is for being rich in the bean powder of organic vanadium and soybean peptides, can directly add seasonings and convert water and drink, also can be used as the v element additive of rich vanadium food, nutritious prod.
The little bean powder of polypeptide of organic vanadium is rich in the preparation of embodiment 4 solid state fermentations
Make raw material with common red bean, or, press the processing method processing and fabricating of embodiment 2 to make raw material by the rich vanadium red bean of embodiment 1 method preparation.
Embodiment 5 liquid fermentation methods prepare rich vanadium soybean polypeptide liquid, rich vanadium soyabean polypeptide powder and rich vanadium Semen sojae atricolor small peptide liquid, rich vanadium Semen sojae atricolor small peptide powder
Make raw material with common soybeans, or make raw material with rich vanadium soybean by embodiment 1 method preparation, pH with 1-7% citric acid or the preparation of other edible acids is the acidic aqueous solution washing of 3.0-7.5 earlier, use flushing with clean water then, 100-125 ℃ was steamed 30-60 minute, 1 part of ripe beans adds the aqueous solution of 5 parts of 0.001-0.006% sodium vanadates, defibrination; Slurries pump into fermentor tank, being heated to 100 ℃ kept 30 minutes, cool 40 ℃, press the 10-60mg/L consumption and add soybean peptides, press the rich vanadium Bacillus subtilus of embodiment 2 method seed selections cultivation and the mixed strains of rich vanadium lactobacillus by the inoculation of 5% inoculum size, wherein rich vanadium Bacillus subtilus and rich vanadium lactobacillus mass ratio are 1: 0.1～0.3; 37-39 ℃ of heat-preservation fermentation in inoculation back 6 hours, viable bacteria is no less than 200,000,000 in every liter of feed liquid of fermenting process maintenance; Fermentation back feed liquid breakdown of emulsion is divided into three layers, and it is liquid going up two-layer, and bottom is a solid-liquid mixing, and the superiors are grease; Separate grease; Remove greasy feed liquid and carry out the screenings separation, gained liquid is the rich vanadium soybean polypeptide liquid that contains a small amount of oligose, and bits are oligose and Mierocrystalline cellulose; Get the rich vanadium soybean peptides of the 0.5-1mol sodium vanadate aqueous solution and 1.5-3mol liquid chelating, concentrate, get the higher rich vanadium soybean polypeptide liquid of content of vanadium, with rich vanadium soybean polypeptide liquid drying then rich vanadium soyabean polypeptide powder, be used to prepare rich vanadium nutritive food, also can be used as the system component for preparing various rich vanadium food.
With above-mentioned rich vanadium soybean polypeptide liquid ultra-filtration membrane and nanofiltration membrane filtering macromole peptide, must make with extra care rich vanadium Semen sojae atricolor small peptide liquid, get the little liquid chelating of the rich vanadium soybean of the 0.5-1mol sodium vanadate aqueous solution and 1.5-3mol again, get the higher rich vanadium Semen sojae atricolor small peptide liquid of vanadiumcontent, remake concentrated, dry, then rich vanadium Semen sojae atricolor small peptide powder, be used to prepare rich vanadium nutritive food, medicine, also can be used as the recipe ingredient for preparing various rich vanadium food.
Embodiment 6 liquid fermentation methods prepare rich vanadium peanut polypeptide liquid, rich vanadium peanut polypeptide powder and the little peptide liquid of rich vanadium peanut, the little peptide powder of rich vanadium peanut
Make raw material with common peanut, or, press the processing method processing and fabricating of embodiment 5 to make raw material by the rich vanadium peanut of embodiment 1 method preparation.
Embodiment 7 prepares rich vanadium soybean polypeptide liquid, rich vanadium soyabean polypeptide powder and rich vanadium Semen sojae atricolor small peptide liquid, rich vanadium Semen sojae atricolor small peptide powder with soybean meal
Make raw material with the dregs of rice that common soybeans squeezes behind the grease, or make raw material with the dregs of rice that squeeze by the rich vanadium soybean of embodiment 1 method preparation behind the grease, get 1 part of soybean meal, pH with 1-7% citric acid or the preparation of other edible acids is the acidic aqueous solution washing of 3.0-7.5, use flushing with clean water then, drain, adding 15-20 part 0.001-0.006% ammonium metavanadate aqueous solution mixes, 100-125 ℃ boiling sterilization 20-60 minute, be cooled to 25-28 ℃, press the 5-30mg/L consumption and add soybean peptides, stir, add the black-koji mould of pressing embodiment 2 method mutagenesis cultivation by 10% inoculum size, also can use the fermented bacterium of cultivating without mutagenesis, 25-28 ℃ fermented 72 hours; The fermentation whole process feeds aseptic compressed air, and constantly stirs, and keeps pH6.5-7.0, as potential of hydrogen unusually then with HCL or Ca (OH)
2Regulate.Carry out screenings after the fermentation and separate, getting supernatant liquor is rich vanadium soybean polypeptide liquid, and its peptide content can reach 60%.Get the rich vanadium soybean polypeptide of 0.5-1mol ammonium metavanadate aqueous solution and 2-3mol liquid chelating, get the higher rich vanadium soybean polypeptide liquid of content of vanadium, get rich vanadium soyabean polypeptide powder through concentrate drying again, be used to prepare rich vanadium nutritive food.
The rich vanadium soybean polypeptide of above-mentioned liquid state is removed the macromole peptide with ultra-filtration membrane and nanofiltration membrane, then rich vanadium Semen sojae atricolor small peptide, get the rich vanadium Semen sojae atricolor small peptide of the 0.5-1mol sodium vanadate aqueous solution and 1.5-3mol liquid chelating again, the higher rich vanadium Semen sojae atricolor small peptide liquid of vanadiumcontent; Rich vanadium Semen sojae atricolor small peptide liquid is concentrated, and drying gets rich vanadium Semen sojae atricolor small peptide powder.
Embodiment 8 prepares rich vanadium peanut polypeptide liquid, rich vanadium peanut polypeptide powder and the little peptide liquid of rich vanadium peanut, the little peptide powder of rich vanadium peanut with peanut meal
Make raw material with the dregs of rice that common peanut is squeezed behind the grease, or make raw material, press the processing method processing and fabricating of embodiment 7 with the dregs of rice that the rich vanadium peanut by the preparation of embodiment 1 method squeezes behind the grease.
Rich vanadium saccharomyces cerevisiae is cultivated in embodiment 9 mutagenesis
Selecting cereuisiae fermentum (Saccharomyces cerevisiae Meyen ex Hansen RCEF1013) to carry out mutagenesis by the method for embodiment 2 cultivates.Wherein, employed substratum is adjusted into its prescription and proportioning,
1. activate substratum: materials and proportioning are 0.001-0.006% ammonium meta-vanadate, 1-3% glucose, 1-3% agar, 0.5-1.5% yeast powder, 0.5-1.5% peptone, soybean peptides 1-6%.
2. seed culture medium: materials and proportioning be, the 0.001-0.006% ammonium meta-vanadate, and 0.8-1.2% glucose, the 0.4-0.6% peptone, the 0.2-0.3% yeast powder, the 15-25%11BX wort, soybean peptides 1-6% adds water to 1000ml.
3. isolation medium: materials and proportioning be, the 0.001-0.006% ammonium meta-vanadate, and 0.8-1.2% glucose, the 0.4-0.6% peptone, the 15-25%11BX wort, soybean peptides 1-6% adds water to 1000ml.
4. fermention medium: materials and proportioning be, the 0.001-0.006% ammonium meta-vanadate, and 0.8-1.2% glucose, the 0.2-0.4% yeast powder, the 0.4-0.6% peptone, the 15-25%11BX wort, soybean peptides 1-6% adds water to 1000ml.
By the mutagenesis of present embodiment method cultivate the rich vanadium cereuisiae fermentum of high vigor, its v element content 〉=800mg/kg (dry weight) gets rich vanadium beer yeast powder with drying, can be directly used in the rich vanadium nutritive food of preparation, also can be used as the system component of the various rich vanadium food of preparation.
Embodiment 10 is a basic raw material with rich vanadium soybean, adds other food raw materials, and preparation is used to modulate the rich vanadium polypeptide of beans sauce
Select rich vanadium soybean 15kg for use, be ground into meal, add hot water 67.5kg, stir 1.471 * 10 by the preparation of embodiment 1 method
3Cook 30min under the Pa pressure;
With flour 4.2kg, salt 5.6kg, water 16kg, mixing stirs evenly the back normal pressure and cooks, preparation seasonings I;
With flour 1.8kg, add water move to 20 ° of B é, add 0.2% calcium chloride and regulate pH to 6.2, add α-Dian Fenmei 3% (every gram raw material adds α-Dian Fenmei 5U) liquefaction, the 100 ℃ of sterilizations that finish of liquefying, be cooled to 60 ℃, add and press rich vanadium black-koji mould (AS3.324) the 7% fermentation 3h that embodiment 2 methods are cultivated, be cooled to 30 ℃, the rich vanadium distillery yeast 5% that embodiment 2 methods are cultivated is pressed in inoculation, normal temperature fermentation 3d makes seasonings II;
With raw material, seasonings I, seasonings II mixing, smash and be cooled to below 50 ℃, press 0.2-0.8% inoculation aspergillus oryzae (UE328 is made in Shanghai), stir water-bath fermentation 15 days.Suitable leavening temperature is 43-47 ℃ of 5 days early stage, 48-50 ℃ of 5 days mid-term, 52-55 ℃ of 5 days later stage.Stirring is once every other day during this time;
Above-mentioned fermentation materials is pressed the 0.001-0.006% consumption add sodium metavanadate, press the 1-3% consumption and add soybean peptides, fully stir, low temperature fermentation one month makes the soybean polypeptide chelating in vanadium ion and the slurries, sterilization, get the slurries of v element content 〉=60mg/kg, be used to modulate beans sauce.
Embodiment 11 is used to modulate the rich vanadium polypeptide of soy sauce with rich vanadium soybean meal preparation
Select the dregs of rice 100kg after the rich vanadium soybean for preparing by embodiment 1 method is squeezed grease for use, with auxiliary material wheat 40kg, wheat bran 10kg mixes, pulverize, adding 95 ℃ of constant temperature of 3 times of amount 33-35% aqueous hydrochloric acids soaked 15 days, adding yellow soda ash is neutralized to about pH7, be cooled to that (be 38-40 ℃ winter about 34-40 ℃, be 34-36 ℃ summer, and spring and autumn is 35-37 ℃), press 0.2-0.5% inoculation aspergillus oryzae (UE336-2 is made in Shanghai), 50 ℃ of ferment at constant temperature 8 days, then cool to 40-45 ℃, press the 0.001-0.006% consumption and add sodium vanadate, press the 1-6% consumption and add soybean peptides, 40-45 ℃ of constant temperature after-ripening 15 days, then cooled to again 33 ℃ of left and right sides ferment at constant temperature 15-20 days, and made the soybean polypeptide chelating in vanadium ion and the slurries, solid-liquid separation, liquid heat to 80 ℃ is kept the 10min sterilization, it is anticorrosion to add an amount of sorbyl alcohol, adds 0.3 odorant % again and increases aquatic foods, adds chitin and stirs companion's removal precipitation, get v element content 〉=80mg/L soybean polypeptide liquid, be used to modulate soy sauce.
Claims (10)
1. method for preparing the peptide that contains vanadium, this peptide that contains vanadium is direct-edible, the material that also can be used as preparation foodstuff additive or medicine, it is characterized in that, select for use the root that is fit to human plant seed edible, rich in protein or plant, stem, leaf as starting material, or be starting material, process by " acidic aqueous solution washing-boiling-interpolation vanadium compound-fermentation-peptide and vanadate chelating " processing step with the soybean meal behind the grease of squeezing out, peanut meal, wherein
Washing is used the edible acid preparation with acidic aqueous solution, and potential of hydrogen is pH3.0-7.5;
Boiling temperature is 100-125 ℃, and cooking time is 20-60 minute;
To spray vanadium compound aqueous solution mode to treating that fermentation materials adds vanadium compound;
Soybean peptides is dissolved in water, is sprayed on and treats to make fermentation accelerant on the fermentation materials;
Cultivated in 20-40 ℃ of temperature 6-72 hour behind the inoculation fermentation bacterium, sterilization is cleaned, and drying is pulverized, the rough vanadium polypeptide powder that contains;
To contain the vanadium polypeptide powder and add the water mill slurry, slagging-off separates and removes not protolysate, gets rich vanadium polypeptide liquid;
Rich vanadium polypeptide liquid is concentrated, and drying is pulverized, and gets rich vanadium polypeptide powder;
Rich vanadium polypeptide liquid is concentrated, and the macromole peptide is removed in ultrafiltration, gets the little peptide liquid of rich vanadium;
The little peptide liquid of rich vanadium is concentrated, and drying is pulverized, and gets the little peptide powder of rich vanadium;
Any peptide class intermediates that contain vanadium with fermentation back gained, liquid direct application, use the solid-state water mill slurry back that then adds earlier, mix with vanadate solution respectively, being adjusted to vanadium peptide mass ratio is 1: the aqueous solution of 2-4, heating, constant temperature, stir, finish chelating, get the higher liquid rich vanadium peptides products of vanadiumcontent;
The rich vanadium peptides products that liquid vanadiumcontent is higher is further done to concentrate, drying treatment, gets the higher rich vanadium peptides products of solid-state vanadiumcontent, comprises liquid rich vanadium polypeptide, solid-state rich vanadium polypeptide, the little peptide of liquid rich vanadium, the little peptide of solid-state rich vanadium.
2. by the described method of claim 1, it is characterized in that, make the suitable mankind plant seed that eat, rich in protein that starting material use and be soybean or peanut.
3. by the described method of claim 1, it is characterized in that, the suitable mankind plant seed that eat, rich in protein of doing the starting material use is, to soybean or peanut do " first vanadium compound, plant growth promoter, seed soak cultivate preservative solution soaking-permanent oxygen of the fixed temperature and humidity of guaranteeing the quality for the first time urge educate-secondary vanadium compound, plant growth promoter, seed soak cultivate preservative solution soaking-secondary guarantee the quality the permanent oxygen of fixed temperature and humidity urge educate-clean, drying " processing, urge for 2 times of gained and to educate rich vanadium soybean or rich vanadium peanut.
4. by the described method of claim 1, it is characterized in that the kind that the acidic aqueous solution that is used for washing material is prepared used edible acid is a kind of of citric acid, oxysuccinic acid, tartrate, acetic acid.
5. by the described method of claim 1, it is characterized in that, add other food raw materials in the material before boiling or after the boiling, in order to regulate the local flavor of the finished product.
6. by the described method of claim 1, it is characterized in that the material fermenting process uses liquid fermentation method, its concrete processing step is, carries out boiling after the material after the washing is broken into slurries with pigment, adds vanadium compound afterwards, stirring makes its dissolving, soybean peptides is added the inoculation fermentation bacterial strain again, in 20-40 ℃ of temperature, cultivated 6-72 hour, stop fermentation, sterilization, drying, pulverize, the rough vanadium polypeptide powder that contains; Liquid material defibrination after maybe will fermenting, slagging-off, not protolysate is removed in separation, gets liquid rich vanadium polypeptide, and further work concentrates, drying treatment, gets solid-state rich vanadium polypeptide.
7. by the described method of claim 1, it is characterized in that the employed zymophyte of fermentation step is any be suitable for protein hydrolysate and edible microorganism.
8. by the described method of claim 7, it is characterized in that the employed zymophyte of fermentation step is bacillus CAU208.
9. by claim 1,7 or 8 described methods, it is characterized in that, the employed fermented bacterium of fermentation step is process ultraviolet mutagenesis or nitrosoguanidine mutagenesis, or through the ultraviolet high vigor bacterial classification of cultivating with the nitrosoguanidine complex mutation, its mutagenesis cultural method is:
(1) preparation contains the vanadium substratum: prepare four kinds of substratum such as activating substratum, seed culture medium, isolation medium, fermention medium respectively with vanadium compound:
1. activate substratum: with peptone, glucose, yeast extract paste, potassium primary phosphate, agar, vanadium compound, soybean peptides and water preparation;
2. seed culture medium: with peptone, glucose, yeast extract paste, potassium primary phosphate, vanadium compound, bean cake powder, soybean peptides and water preparation;
3. isolation medium: with peptone, glucose, yeast extract paste, potassium primary phosphate, skim-milk, agar, vanadium compound, soybean peptides and water preparation;
4. fermention medium: with bean cake powder, peptone, molasses, vanadium compound, soybean peptides and water preparation;
(2) fermented bacterium mutagenesis:
Ultraviolet mutagenesis: bacterial classification is after activating the cultivation of substratum and seed culture medium, with UV-lamp irradiation mutagenesis;
Nitrosoguanidine mutagenesis: bacterial classification adds nitrosoguanidine mutagenesis through activating after substratum and seed culture medium are cultivated;
Ultraviolet ray and nitrosoguanidine complex mutation: bacterial classification adds nitrosoguanidine earlier after activation substratum and seed culture medium cultivation, the UV-lamp of going is again shone mutagenesis;
(3) strain screening:
Through ultraviolet mutagenesis, or nitrosoguanidine mutagenesis, or primary dcreening operation is just being become bacterial strain in the zymophyte behind ultraviolet ray and the nitrosoguanidine complex mutation; The primary dcreening operation gained is just become bacterial strain inserting in the fermention medium and cultivate, measuring the degree of hydrolysis of protein hydrolyzate, multiple sieve, just change bacterial strain that must be more stable; Further after cultivating in the isolation medium, the transparent circle diameter in the mensuration isolation medium and the ratio of colony diameter, filter out the big bacterial strain of ratio, cultivate through seed culture medium, be inoculated in the fermention medium, measure the fermented liquid degree of hydrolysis behind certain fermentation period, just change bacterial strain that must be more stable is divided into two groups of 2 multiple sieves, and one group is carried out UV treatment, one group is carried out nitrosoguanidine and handles, select wherein functional and stable bacterial strain and carry out ultraviolet ray and nitrosoguanidine mutagenesis simultaneously, get the superior strain of excellent in stability, superior strain is inoculated into activates on the substratum as the 1st generation, went down to posterity 1 time every 5 days later on, passed for 10 generations altogether, each for shake flask fermentation, is measured the degree of hydrolysis of fermented liquid, selected good stability, high yield is good, the regressive bacterial strain of not undergoing mutation keeps, the rich vanadium fermentation strain of promptly high vigor.
10. by the product of claim 1～4 and the described method preparation of 6～9 arbitrary claims, it is characterized in that with dry weight basis, wherein peptide content is more than 60%, v element content is more than 100mg/kg.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104738384A (en) * | 2014-08-27 | 2015-07-01 | 何定 | Preparation method of hydrolyzed protein peptide product |
| CN105815565A (en) * | 2016-03-28 | 2016-08-03 | 宋天文 | Sugar factory waste liquid treatment method |
| CN106578961A (en) * | 2016-12-13 | 2017-04-26 | 黑龙江省农业科学院食品加工研究所 | Method for preparing red bean peptide oral liquid with effect of reducing blood pressure and blood glucose by using fermentation method |
| CN113749175A (en) * | 2021-09-09 | 2021-12-07 | 中新国际联合研究院 | Preparation method of bitter-free soybean polypeptide with blood fat reducing activity |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0328157D0 (en) * | 2003-12-04 | 2004-01-07 | Imp College Innovations Ltd | Compounds |
| GB0421836D0 (en) * | 2004-10-01 | 2004-11-03 | Avidex Ltd | T cell receptors containing a non-native disulfide interchain bond linked to immunomodulatory agents |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104738384A (en) * | 2014-08-27 | 2015-07-01 | 何定 | Preparation method of hydrolyzed protein peptide product |
| CN105815565A (en) * | 2016-03-28 | 2016-08-03 | 宋天文 | Sugar factory waste liquid treatment method |
| CN106578961A (en) * | 2016-12-13 | 2017-04-26 | 黑龙江省农业科学院食品加工研究所 | Method for preparing red bean peptide oral liquid with effect of reducing blood pressure and blood glucose by using fermentation method |
| CN113749175A (en) * | 2021-09-09 | 2021-12-07 | 中新国际联合研究院 | Preparation method of bitter-free soybean polypeptide with blood fat reducing activity |
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