CN104788533B - A kind of method for producing oligopeptides and amino acid from high temperature grouts using subcritical water - Google Patents
A kind of method for producing oligopeptides and amino acid from high temperature grouts using subcritical water Download PDFInfo
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Abstract
The invention belongs to fats and oils processing accessory substance deep process technology field, and in particular to a kind of method for producing oligopeptides and amino acid from high temperature grouts using subcritical water, be specially：First by being heated to reflux reclaiming the residual grease in high temperature grouts, then using subcritical water hydrolysis grouts, through centrifuging, decolourizing, ultrafiltration, nanofiltration separation, spray drying obtain oligopeptides and amino acid respectively.Compared with traditional chemical hydrolysis preparation method, the inventive method has the advantages that product quality is high, production cost is low, green energy conservation.The inventive method is both realized comprehensive utilization of resources, is increased economic well-being of workers and staff again, have positive social and economic significance using the oligopeptides and amino acid products of fats and oils processing accessory substance high temperature grouts production high added value.
Description
Technical field
The inventive method belongs to the deep process technology field of fats and oils processing accessory substance, and in particular to one utilizes kind of a subcritical water
The method that production oligopeptides and amino acid are prepared from high temperature grouts.
Background technology
China is production and consumption big country, China's rapeseed, soybean, peanut, cottonseed, the sunflower in 2013 of edible oil and fat
Seed, sesame, tea seed and the big oil plant total output of linseed eight are 5846.8 ten thousand tons.Due to oil plant, supply falls short of demand, and China is entered in the same year
Mouth oil plant reaches 2453.9 ten thousand tons.This year China's edible oil and fat supply is 3374.3 ten thousand tons, 22.5 kilograms of pre-capita consumption grease.
Substantial amounts of grouts can be produced during edible fat production, by taking soybean cake dregs as an example, 2013 annual output soybean cake dregs are about 4483.5 ten thousand
Ton.Research shows：Rich in functional components such as albumen, polysaccharide in oil meal.Therefore, separation egg has been produced both at home and abroad
In vain, feed etc..Influenceed by China's consumption of resident custom and existing processing technology, high-temperature baking flavouring and high temperature squeezing are edible
The common technology of grease production, thus produce high temperature grouts.High temperature grouts are one kind of grouts, and it is produced by following two techniques more
It is raw：First, the oil-producing technique for coordinating high-pressure compressing by 100 DEG C of temperatures above produces；Second, there is 160-200 DEG C of high temperature to bake
The oil-producing technique of oil plant Sheng Xiang workshop sections produces.High temperature action can cause oilseed protein to be denatured, and reduce or lose original feature,
So as to limit the subsequent applications of oilseed protein.At present, substantial amounts of high temperature grouts are used as animal feed or fertilizer, added value of product
Low, resource is underutilized.Therefore, it is necessary to carry out the deep processing research of high temperature grouts.
In order to prepare oligopeptides and amino acid using enzymatic hydrolysis, chemical degradation using albumen in high temperature grouts, researcher.
Compared to albumen, oligopeptides is more easy to assimilate, and with functions such as anti-oxidant, removing free radical, regulation blood pressures.Amino acid except
It is beyond the base substance for forming human body, also there is treatment hepatopathy, disease of digestive tract, improve the functions such as muscular strength.Therefore both at home and abroad
There is Related product sale, and obtained consumer's accreditation, the bright outlook with the market development.However, Production by Enzymes polypeptide
Cost is higher, and single enzymolysis polypeptide is low, and the less stable of enzyme addition, which has limited enzyme process to prepare polypeptide and amino acid
Industrialization conversion.Chemical degradation can degrade oligopeptides and amino acid quality, and be polluted the environment in production process, and power consumption is higher,
This also limits chemically treated development.
Subcritical water refers to temperature between 100 DEG C and critical-temperature（374℃）Between, pressure is less than critical pressure（22.4
MPa）, and density is higher than critical density（0.32 g/cm3）Aqueous water.Because HTHP causes hydrogen bond in water, ionized water
The microstructures such as conjunction, tufted structure change, so as to cause subcritical water to have low-k and strong catalytic capability.Because Asia is faced
Boundary's water technology makees reaction medium using pure water, reduces the usage amount of organic reagent and acid base catalysator, simplifies follow-up organic molten
Separating technology is reclaimed in agent.This both reduces production cost, decreases environmental pollution, therefore subcritical water is degraded high temperature grouts system
Standby oligopeptides has researching value with amino acid.
The content of the invention
Present invention aims at providing a kind of method for producing oligopeptides and amino acid from high temperature grouts using subcritical water,
It is high using this method production oligopeptides and amino acid purity height, simple production process, added value of product.
To achieve the above object, the technical scheme that the present invention takes is as follows：
A kind of method for producing oligopeptides and amino acid from high temperature grouts using subcritical water, it comprises the following steps：
1）High temperature grouts pre-process：It will crush（Typically being crushed to 16-60 mesh is advisable）, high temperature grouts after sieving are with having
Residual oil in solvent heating and refluxing extraction grouts, backflow centrifuge after terminating（With 2000-5000 rpm centrifugation 10-30 min
It is advisable）, precipitation is collected, is evaporated under reduced pressure and removes residual solvent, it is standby；
Vacuum distillation condition is preferred：Pressure -0.08--0.10 MPa, temperature are 60-90 DEG C, time 10-30 min；
2）Subcritical water processing：By step 1）The precipitation that gained removes residual solvent mixes with water, is made into 5-20wt%'s
PH is simultaneously adjusted to 3.0-7.0 by solution, is then transferred in high-temperature high-pressure reaction kettle, in 50-120 rpm stirring condition
Under in 120-280 DEG C of hydrolysis 10-70 min, after hydrolysis terminates, be cooled to room temperature；
3）Hydrolyzate purification process：Take step 2）Gained reaction system, pH to 4.1-4.6 is adjusted, is then centrifuged
（It is advisable with 4000-5000 rpm centrifugations 10-40 min）And supernatant is collected, supernatant is decolourized with macroreticular resin, filtering,
Ultrafiltration remove high molecular weight protein, neutralize ultrafiltration permeate, NF membrane desalination, after desalination solution then with NF membrane isolated polypeptide with
Amino acid, after polypeptide liquid and aminoacids solution are concentrated, oligopeptides and amino acid dry powder are obtained using spray drying.
Wherein, step 3）In, macroreticular resin decolorization condition is specially：At 20-40 DEG C, macroreticular resin and hydrolyzate volume
Than for 1:5-25,30-120 rpm stirring 30-90 min.Ultrafiltration removes high molecular weight protein condition：In 20-40
DEG C, operating pressure 0.15-0.35 MPa, solid concentration is 3-6% in feed liquid.Nanofiltration desalination condition is specially：Operating pressure
For 0.20-0.40 MPa, solid concentration is 2-5 % in feed liquid.The condition of nanofiltration separation polypeptide and amino acid is specially：
Operating pressure 0.20-0.40 MPa, solid concentration is 2-5% in feed liquid.Pressuresprayingdrier, 5- are selected in spray drying
9% solid concentration, 160-180 DEG C of inlet temperature, outlet temperature are 70-80 DEG C, and pressure is 1.2-1.6 MPa.
Specifically, step 1）In, the high temperature grouts be high temperature sesame cake meal, high temperature peanut grouts, high temperature soybean cake dregs,
High temperature dregs of rapeseed cake or high temperature cotton cake dregs.The organic solvent is petroleum ether, pentane or ether.Heated reflux condition is：
60-90 DEG C of heating-up temperature, is heated to reflux time 1-3 h, and the amount ratio of high temperature grouts and organic solvent is 1g：8-20ml.
Step 2）In, it is acetic acid, phosphoric acid, hydrochloric acid or sulfuric acid to adjust the acid used in pH, and alkali used is sodium hydroxide, hydrogen-oxygen
Change potassium or sodium carbonate.
Step 2）Middle pH is adjusted to 3.0-7.0, if product is using amino acid to be main, pH is adjusted to 3-5；If product is with oligopeptides
For it is main when, then pH is adjusted to 5-7.
Step 2）Middle 120-280 DEG C of hydrolysising reacting temperature, if product is using amino acid to be main, hydrolysising reacting temperature is adopted
With 200-280 DEG C；If product is using oligopeptides to be main, hydrolysising reacting temperature uses 120-200 DEG C.
Step 3）Middle big pore resin is D301-G, D354 or D370 weakly basic styrene type anion exchange resin.
Milipore filter used in retention albumen uses milipore filter of the molecular cut off for 8000-10000 dalton；NF membrane is adopted used in desalination
With the NF membrane that molecular cut off is 70-100 dalton；Isolated polypeptide uses molecular cut off with NF membrane used in amino acid
For the NF membrane of 200-220 dalton.
The invention provides a kind of oligopeptides for preparing high added value from high temperature grouts using subcritical water and amino acid
Production technology, this method more traditional strong acid hydrolysis method, both reduce chemical reagent using subcritical water technology hydrolysis high temperature grouts
Usage amount, also shorten the production time, production cost is low；The oligopeptides and ammonia that UF membrane, resin decolorization and spray drying obtain
Base acid purity height and quality better.The inventive method improves the utilization rate of Oil processing accessory substance grouts, can be not only social band
Carry out abundant economic value, and the oligopeptides with physiological hygiene and trophic function is advantageous to improve China residence with amino acid products
The fitness of the people, there is positive social value.
Compared with prior art, the inventive method has the beneficial effect that：
1）Subcritical hydrolysis prepares oligopeptides simple with amino acids production technique, green energy conservation, production efficiency height, easily realized
Industrialization；
2）Active peptide powder is prepared using spray drying, on the one hand improves and shortens drying time, improves production efficiency, it is another
Aspect improves the rehydration dissolubility of Gly-His-Lys；
3）To feed liquid isolate and purify desalination using membrane separation technique at normal temperatures in the present invention, it is energy-efficient and without dirt
Dye；
4）Oligopeptides produced by the invention and amino acid purity height, quality better, can be widely applied to cosmetics, food, medicine
Deng field.
Embodiment
The present invention is described in further details below by way of preferred embodiment, but protection scope of the present invention is not limited to
In this.
Used related assays method is as follows in following each embodiments：
1. the assay method of oligopeptides purity：Using kjeldahl determination（GB 5009.5-2010）.
2. trichloroacetic acid nitrogen soluble index assay method：
Take respectively two parts it is identical in quality（1g）Peptide dry powder, by a copy of it with 10mL distilled water dissolve, then add 10
The solution of trichloroacetic acid of mL concentration 10% mixes, and stands 30min, 3500rpm centrifugation 5min, supernatant is calculated using Kjeldahl's method
The quality (g) of soluble nitrogen and the quality (g) of total nitrogen in portion in addition in liquid.Calculation formula is as follows：
Trichloroacetic acid nitrogen soluble index（%）=(nitrogen quality/total nitrogen quality in supernatant) × 100
Trichloroacetic acid nitrogen soluble index is higher, shows that the impurity contents such as albumen in Gly-His-Lys, polypeptide are lower, peptide powder purity is got over
It is high.
3. amino acid composition measuring：Determined using automatic amino acid analyzer（GB/T 5009.124-2003）.
Embodiment 1：
A kind of method for producing oligopeptides and amino acid from high temperature grouts using subcritical water, it comprises the following steps：
1）High temperature sesame cake meal pre-processes：
100 g are taken to crush（It is crushed to 20 mesh）, high temperature sesame cake meal after sieving, be transferred to 2000 mL round-bottomed flasks
In, add 1000 mL pentanes, 80 DEG C of stirrings（80 rpm）3 h are heated to reflux, 4000 rpm centrifugations 25 after backflow terminates
Min, collect precipitation.Precipitation is poured into round-bottomed flask, 80 DEG C, -0.10 MPa vacuum distillation 20 min removing residual solvents, received
It is standby to collect solid.
2）Subcritical water processing：
Take step 1）The precipitation that gained removes residual solvent mixes with water, is made into 10wt% solution, and with 5 mol/L's
PH is adjusted to 5.0 by HCl, and then the solution is transferred in high-temperature high-pressure reaction kettle, in 200 DEG C under 100 rpm stirring conditions
The min of hydrolysis 40.After hydrolysis terminates, room temperature is cooled to.
3）Hydrolyzate purification process：
Take step 2）The reaction system of gained, pH to 4.3 is adjusted, subsequent 5000 rpm centrifuges 30 min, collects supernatant.
Supernatant D301-G macroreticular resins decolourize, and decolorization condition is：The ratio of macroreticular resin and hydrolyzate is 1:90 at 10,30 DEG C
Rpm stirs 60 min.Filtering.Milipore filter of the molecular cut off for 8000 dalton then is used, is gone with 0.20MPa operating pressures
Except high molecular weight protein.After ultrafiltration permeate is neutralized, the NF membrane for using molecular cut off to be 100 dalton is grasped with 0.25 MPa
Make 10 times of desalinations of pressure dilution.Solution uses NF membrane isolated polypeptide and amino of the molecular cut off for 200 dalton after desalination
Acid.Polypeptide liquid is 6% with rotated be concentrated by evaporation to concentration of aminoacids solution, and oligopeptides and amino acid dry powder are obtained using spray drying.
The 15.7 g sesame oligopeptides dry powder obtained are a kind of off-white powders, and in the case of butt, peptide purity is 94.7%, ash
It is divided into 0.5%, sugared content 4.5%, fat content 0.3%；Trichloroacetic acid nitrogen soluble index is 99%.In addition, obtain 12.4 g
Amino acid products, the composition of amino acid products are shown in Table 1.
Embodiment 2：
A kind of method for producing amino acid and oligopeptides from high temperature peanut grouts using subcritical water, it includes following step
Suddenly：
1）High temperature peanut grouts pre-process：
200 g are taken to crush（It is crushed to 30 mesh）, high temperature peanut grouts after sieving, be transferred in 5000 mL round-bottomed flasks,
It is 30-60 DEG C of petroleum ether to add 3000 mL boiling ranges, and 70 DEG C are stirred（100 rpm）2 h are heated to reflux, backflow terminates rear 5000
Rpm centrifuges 30 min, collects precipitation.Precipitation is poured into round-bottomed flask, 70 DEG C, -0.10 MPa vacuum distillation 20 min removings
Residual solvent, it is standby to collect solid.
2）Subcritical water processing：
Take step 1）The precipitation that gained removes residual solvent mixes with water, is made into 8wt% solution, and with 5 mol/L's
H2SO4PH is adjusted to 4.5, then the solution is transferred in high-temperature high-pressure reaction kettle, in 220 DEG C under 90 rpm stirring conditions
The min of hydrolysis 60.After hydrolysis terminates, room temperature is cooled to.
3）Hydrolyzate purification process
Take step 2）The reaction system of gained, pH to 4.5 is adjusted, subsequent 5000 rpm centrifuges 30 min, collects supernatant.
Supernatant D370 macroreticular resins decolourize, and decolorization condition is：The ratio of macroreticular resin and hydrolyzate is 1:90 rpm at 8,30 DEG C
Stir 80 min.Filtering.Milipore filter of the molecular cut off for 10000 dalton then is used, is removed with 0.22MPa operating pressures
High molecular weight protein.After ultrafiltration permeate is neutralized, the dalton NF membrane of molecular cut off 100 is used with 0.25 MPa operating pressures
Dilute 15 times of desalinations.Solution uses NF membrane isolated polypeptide and amino acid of the molecular cut off for 200 dalton after desalination.Polypeptide
Liquid is 8% with rotated be concentrated by evaporation to concentration of aminoacids solution, and oligopeptides and amino acid dry powder are obtained using spray drying.
The 15.1g peanut oligopeptides dry powder of acquisition is a kind of off-white powder, and in the case of butt, peptide purity is 93.9%, ash content
For 0.5%, sugared content 5.3%, fat content 0.3%；Trichloroacetic acid nitrogen soluble index is 99%.In addition, obtain 28.6 g ammonia
Base acid product, the composition of amino acid products are shown in Table 2.
Embodiment 3：
A kind of method for producing amino acid and oligopeptides from high temperature peanut grouts using subcritical water, it includes following step
Suddenly：
1）High temperature peanut grouts pre-process：
200 g are taken to crush（It is crushed to 20 mesh）, high temperature peanut grouts after sieving, be transferred in 5000 mL round-bottomed flasks,
Add 3000 mL ether, 60 DEG C of stirrings（100 rpm）3h is heated to reflux, 4000 rpm centrifuge 30 min after backflow terminates, and collect
Precipitation.Precipitation is poured into round-bottomed flask, 60 DEG C, -0.10 MPa vacuum distillation 20 min removing residual solvents, it is standby to collect solid
With.
2）Subcritical water processing：
Take step 1）The precipitation that gained removes residual solvent mixes with water, is made into 10wt% solution, and with 5 mol/L's
H3PO4PH is adjusted to 6.0, then the solution is transferred in high-temperature high-pressure reaction kettle, in 160 DEG C under 90 rpm stirring conditions
The min of hydrolysis 30.After hydrolysis terminates, room temperature is cooled to.
3）Hydrolyzate purification process
Take step 2）The reaction system of gained, pH to 4.5 is adjusted, subsequent 4500 rpm centrifuges 30 min, collects supernatant.
Supernatant D354 macroreticular resins decolourize, and decolorization condition is：The ratio of macroreticular resin and hydrolyzate is 1:100 at 15,30 DEG C
Rpm stirs 90 min.Filtering.Milipore filter of the molecular cut off for 10000 dalton then is used, with 0.25MPa operating pressures
Remove high molecular weight protein.After ultrafiltration permeate is neutralized, the dalton NF membrane of molecular cut off 100 is used to be operated with 0.30 MPa
18 times of desalinations of pressure dilution.Solution uses NF membrane isolated polypeptide and amino acid of the molecular cut off for 200 dalton after desalination.
Polypeptide liquid is 5% with rotated be concentrated by evaporation to concentration of aminoacids solution, and oligopeptides and amino acid dry powder are obtained using spray drying.
The 28.9g peanut oligopeptides dry powder of acquisition is a kind of off-white powder, and in the case of butt, peptide purity is 94.1%, ash content
For 0.3%, sugared content 5.3%, fat content 0.3%；Trichloroacetic acid nitrogen soluble index is 99%.In addition, obtain 16.1 g ammonia
Base acid product, the composition of amino acid products are shown in Table 3.
Claims (3)
- A kind of 1. method for producing oligopeptides and amino acid from high temperature grouts using subcritical water, it is characterised in that including as follows Step：1) high temperature grouts pre-process：It will crush, the high temperature grouts after sieving are with residual in organic solvent heating and refluxing extraction grouts Oil, backflow centrifuge after terminating, and collect precipitation, are evaporated under reduced pressure and remove residual solvent, standby；2) subcritical water is handled：The precipitation that residual solvent is removed obtained by step 1) is mixed with water, is made into 5-20wt% solution And pH is adjusted to 3.0-7.0, be then transferred in high-temperature high-pressure reaction kettle, under 50-120rpm stirring condition in 120-280 DEG C of hydrolysis 10-70min, after hydrolysis terminates, is cooled to room temperature；3) hydrolyzate purification process：Reaction system obtained by step 2) is taken, pH to 4.1-4.6 is adjusted, is then centrifuged and collected Supernatant, supernatant are decolourized with macroreticular resin, and ultrafiltration removes high molecular weight protein, neutralize ultrafiltration permeate, NF membrane desalination, desalination Solution then uses NF membrane isolated polypeptide and amino acid afterwards, after polypeptide liquid and aminoacids solution are concentrated, is obtained using spray drying Oligopeptides and amino acid dry powder；PH is adjusted to 3.0-7.0 in step 2), if product is using amino acid to be main, pH is adjusted to 3-5；If product is based on oligopeptides When, pH is adjusted to 5-7；120-280 DEG C of hydrolysising reacting temperature in step 2), if product is using amino acid to be main, hydrolysising reacting temperature uses 200- 280℃；If product is using oligopeptides to be main, hydrolysising reacting temperature uses 120-200 DEG C；Big pore resin is D301-G, D354 or D370 weakly basic styrene type anion exchange resin in step 3)；Retention Milipore filter used in albumen uses milipore filter of the molecular cut off for 8000-10000 dalton；NF membrane used in desalination, which uses, cuts Stay the NF membrane that molecular weight is 70-100 dalton；Isolated polypeptide and NF membrane used in amino acid use molecular cut off for The NF membrane of 200-220 dalton.
- 2. the method that oligopeptides and amino acid are produced from high temperature grouts using subcritical water as claimed in claim 1, its feature exist In in step 1), the high temperature grouts are high temperature sesame cake meal, high temperature peanut grouts, high temperature soybean cake dregs, high temperature dregs of rapeseed cake Or high temperature cotton cake dregs；The organic solvent is petroleum ether, pentane or ether；Heated reflux condition is：Heating-up temperature 60- 90 DEG C, it is heated to reflux time 1-3h, the amount ratio of high temperature grouts and organic solvent is 1g：8-20ml.
- 3. the method that oligopeptides and amino acid are produced from high temperature grouts using subcritical water as claimed in claim 1, its feature exist In in step 2), it is acetic acid, phosphoric acid, hydrochloric acid or sulfuric acid to adjust the acid used in pH, and alkali used is sodium hydroxide, potassium hydroxide Or sodium carbonate.
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| CN102060936A (en) * | 2011-01-05 | 2011-05-18 | 江苏大学 | Method for extracting rice bran polysaccharide from sub-critical water |
| CN102276665A (en) * | 2011-06-21 | 2011-12-14 | 晨光生物科技集团天津有限公司 | Method of extracting flax lignin from flax seed cake |
| CN103005141A (en) * | 2012-12-17 | 2013-04-03 | 南昌大学 | Method for improving oxidation resistance and functional property of plant protein |
| CN103880972A (en) * | 2014-03-12 | 2014-06-25 | 江苏大学 | Method of synchronously extracting polysaccharides and proteins from subcritical water |
| CN104286370A (en) * | 2014-08-20 | 2015-01-21 | 华南理工大学 | Method for preparing functional soybean protein by enzyme-assistant subcritical water method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102060936A (en) * | 2011-01-05 | 2011-05-18 | 江苏大学 | Method for extracting rice bran polysaccharide from sub-critical water |
| CN102276665A (en) * | 2011-06-21 | 2011-12-14 | 晨光生物科技集团天津有限公司 | Method of extracting flax lignin from flax seed cake |
| CN103005141A (en) * | 2012-12-17 | 2013-04-03 | 南昌大学 | Method for improving oxidation resistance and functional property of plant protein |
| CN103880972A (en) * | 2014-03-12 | 2014-06-25 | 江苏大学 | Method of synchronously extracting polysaccharides and proteins from subcritical water |
| CN104286370A (en) * | 2014-08-20 | 2015-01-21 | 华南理工大学 | Method for preparing functional soybean protein by enzyme-assistant subcritical water method |
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