CN108504709A - A kind of maize oligopeptide and its industrialized preparing process - Google Patents
A kind of maize oligopeptide and its industrialized preparing process Download PDFInfo
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- CN108504709A CN108504709A CN201810262531.6A CN201810262531A CN108504709A CN 108504709 A CN108504709 A CN 108504709A CN 201810262531 A CN201810262531 A CN 201810262531A CN 108504709 A CN108504709 A CN 108504709A
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Abstract
A kind of maize oligopeptide of present invention offer and its industrialized preparing process wherein include at least peptide fragment pEL, VL and LL in maize oligopeptide composition.The industrialized preparing process of the maize oligopeptide, includes the following steps：Maize yellow-powder is implemented to digest using protease, obtains enzymolysis liquid；Implement centrifugation, filtering, activated carbon adsorption, cation exchange resin purifying and anion exchange resin purification process successively to enzymolysis liquid.Using industrialized preparing process provided by the present invention, pEL with different physiological roles such as protect liver, anti-inflammatory and antidepressions can be retained, as these three functional peptide fragments of the VL and LL of good branched-chain amino acid propylhomoserin replenishers.
Description
Technical field
The present invention relates to a kind of maize oligopeptide and its industrialized preparing process, more particularly to one kind to have pyroglutamyl bright
The maize oligopeptide and its industrialized producing technology of propylhomoserin, valyl leucine and leucyl leucine peptide fragment.
Background technology
Corn is one of Three major grain crops and China's conventional crop in the world.One of main application of corn is
Produce cornstarch, wet grinding production cornstarch during principal by product be corn protein powder (CornGlutenMeal,
CGM), also referred to as maize yellow-powder.Corn protein powder generally contain 60% or so protein, main component be zeins,
Glutelin and globulin.Contain the hydrophobicitys such as a high proportion of leucine, isoleucine, valine, alanine in corn protein powder
Amino acid and proline, glutamic acid etc., but the essential amino acids contents such as lysine, tryptophan are relatively low, this unique amino acid
Composition keeps zein poorly water-soluble, nutritive value not high.
Using Protease Treatment corn protein powder, after so that it is hydrolyzed into the maize oligopeptide of small molecule, can be completely dissolved
Yu Shui, to make it possible its application in food.The study found that maize oligopeptide is rich in branched-chain amino acid, have
Promote protein synthesis in muscle and inhibit the effect of breaks down proteins, can be used for arsenic and clinical nutrition product；It is beautiful
Rice oligopeptide contains a large amount of alanine, the speed that body absorbs ethyl alcohol can be made to slow down, and can promote alcohol metabolism, reduces its poison
Property, caused acute alcoholism of drinking can be greatly reduced.It reduces blood pressure in addition, maize oligopeptide has, reduce blood fat, it is auxiliary
Help the physiological activity for the treatment of hepatopathy and breast cancer.
Maize oligopeptide is usually using corn protein powder as raw material, through the techniques system such as sizing mixing, digesting, be separated, purified, dried
.In current production technology, the purity for focusing on maize oligopeptide and oligomeric peptide content more, and retain dipeptides as much as possible
And tripeptides, so that it is more advantageous to absorption of human body.For example in patent CN201410357708.2 it is exactly by using alkali protease
The corn protein powder after degreasing decoloring is hydrolyzed with neutral proteinase, then passes through series of separate purification process, to obtain
Oligopeptide as much as possible；A kind of preparation process of maize oligopeptide is just described in patent CN200810084992.5, is made low
The molecular weight distribution of poly- peptide is in 2000Da or less.
But in the production process of maize oligopeptide, in order to retain the oligopeptides such as more dipeptides, tripeptides as far as possible
Or oligopeptide purity is pursued, some important functional peptide fragments, such as burnt paddy ammonia are often lost in enzymolysis liquid purification process
Acyl leucine (pyroGlu-Leu, pEL), valyl leucine (Vla-Leu, VL), leucyl leucine (Leu-Leu, LL) etc.
Three peptide fragments.And the result of study that Yukako Yamamoto et al. were published in 2015 on periodical Neuropeptides
《Antidepressant-like effect of food-derived pyroglutamyl peptides in mice》In
Show that pEL has the function of that protect liver, other researchs also indicate that, pEL also has anti-inflammatory and antidepressant different physiological roles；
And VL and LL are also it is verified that good branched-chain amino acid propylhomoserin replenishers can be used as.But it is produced in existing maize oligopeptide
In product, it is substantially not detectable above-mentioned functional peptide fragment.
Invention content
In view of the above-mentioned drawbacks in the prior art, the present invention provides a kind of maize oligopeptide, retains in the maize oligopeptide
Pyroglutamyl leucine (pyroGlu-Leu, pEL), valyl leucine (Vla-Leu, VL), leucyl leucine (Leu-
Leu, LL) these three functional peptide fragments.
The present invention also provides the industrialized preparing process of above-mentioned maize oligopeptide to be made by implementing specific purifying process
PEL with different physiological roles such as protect liver, anti-inflammatory and antidepressions and as good branched-chain amino acid propylhomoserin replenishers
VL and LL these three functional peptide fragments retained.
To achieve the above object, present invention firstly provides a kind of maize oligopeptide, which at least wraps in forming
Include peptide fragment pEL, VL and LL.
Specifically, in maize oligopeptide provided by the present invention, the gross mass (butt) based on maize oligopeptide, burnt paddy ammonia
Content >=0.85% of acyl leucine (pEL), content >=0.15% of valyl leucine (VL), leucyl leucine (LL)
Content >=1.15%.
According to maize oligopeptide provided by the invention, be by maize yellow-powder carry out protease hydrolyzed after, then through point
Made from purifying,
Wherein, above-mentioned isolate and purify includes implementing centrifugation, filtering, activated carbon adsorption, cation exchange to enzymolysis liquid successively
Purifying resin and anion exchange resin purifying.
Above-mentioned protease can be specifically the two steps enzymolysis that alkali protease is combined with neutral proteinase, usually first use
Alkali protease digests maize yellow-powder, continues to digest using neutral proteinase again after enzymolysis, go out after enzymolysis
Then enzyme implements subsequent separation and purification treatment to obtained enzymolysis liquid.
It is to mix maize yellow-powder with water, such as by maize yellow-powder and deionized water in specific implementation process of the present invention
According to 1：The mass ratio mixing of (5-20) is sized mixing, and alkali protease is then added and is digested, wherein adding per 100g maize yellow-powders
The alkali protease entered is 1.85-2.95AU, and neutral proteinase is then added and is digested, wherein being added per 100g maize yellow-powders
Neutral proteinase be 0.45-0.90AU.
The specific addition of alkali protease and neutral proteinase can rationally be adjusted according to its vigor, for example be had in the present invention
In body implementation process, used alkali protease believes (China) Bioisystech Co., Ltd purchased from Novi, specially
About 0.9g alkali proteases are added per 100g maize yellow-powders in Alcalase 2.4L type alkali proteases；Used neutral protein
Enzyme believes (China) Bioisystech Co., Ltd, specially Neutrase 0.8L types neutral proteinase purchased from Novi, per 100g corns
About 0.72g neutral proteinases are added in bloom.
Hydrolysis temperature and enzymolysis time can determine according to the actual conditions of alkali protease used and neutral proteinase, such as
The hydrolysis temperature of alkali protease can be controlled at 50-65 DEG C, and it is 8-10 to adjust pH value, and enzymolysis time is 20min or more, i.e.,
Effectively enzymolysis can be achieved, enzymolysis time is long to generate too much influence, the enzymolysis of general alkali protease to special peptide fragment
Time is 20-60min, such as 30min.The hydrolysis temperature of neutral proteinase can specifically be controlled at 50-65 DEG C, enzymolysis time control
System generally can be controlled in 150-240min, such as 210min in 150min or more.
After the completion of enzymolysis, this field conventional means can be used and carry out enzyme deactivation, for example is warming up to 100 DEG C and maintains 30min left
It is right.
Fail the solid impurity of enzymolysis in enzymolysis liquid containing part, can be removed by centrifugation, in actual industrial production
In, the solid impurity in decanter centrifuge (horizontal type screw settling centrifuge) removal enzymolysis liquid generally can be used；In scale up test
Or when small-scale production, desk centrifuge progress generally can be used.
After centrifugal treating, obtained centrifugate is clarified substantially, then can be further removed by filtration therein big
Molecular substance, the ceramic membrane that the apertures 50-200nm generally can be used implement micro-filtration, then use activated carbon adsorption with hardship of decolourizing
And aromatic amino acid is removed, and through cation exchange resin and anion exchange resin desalination and remove free amino acid successively
Etc. charged species, by above-mentioned separating-purifying, these three functional peptide fragments of pEL, VL and LL in enzymolysis liquid are retained.
Product after separating-purifying is through being dried, you can obtains powdered maize oligopeptide, i.e. corn oligopeptide powder.
The present invention also provides the industrialized preparing process of above-mentioned maize oligopeptide, include the following steps：
Maize yellow-powder is implemented to digest using protease, obtains enzymolysis liquid；
Implement centrifugation, filtering, activated carbon adsorption, cation exchange resin purifying and anion successively to the enzymolysis liquid to hand over
Change purifying resin processing.
According to the industrialized preparing process of maize oligopeptide provided by the invention, in purifying technique, centrifuge for removing
Solid impurity in enzymolysis liquid, filtering can remove macromolecular substances therein, activated carbon adsorption hardship and is removed for decolourizing
The charged species such as aromatic amino acid, anion-cation exchange resin desalination and removal free amino acid, in the process, pEL,
These three functional peptide fragments of VL and LL are retained.
The present invention is not specially limited maize yellow-powder raw material used, use common maize yellow-powder production currently on the market
Product, protein content therein are not less than 50%, are generally concentrated at 50%-60%, for example be purchased from Qinhuangdao black horse Hua starch
60 type maize yellow-powders of limited liability company.
The present invention is not specially limited specific enzymolysis process, and the common enzyme of maize oligopeptide institute is prepared using this field
Technique is solved, protein therein is fully hydrolyzed to oligopeptide and acquisition peptide fragment pEL, VL and LL as much as possible.
In specific implementation process of the present invention, the concrete technology of taken enzymolysis includes：
After maize yellow-powder is mixed with water, sequentially adds alkali protease and neutral proteinase is digested, last enzyme deactivation
Processing, obtains the enzymolysis liquid, wherein：
The mass ratio of maize yellow-powder and water is 1：(5-20)；The alkali protease and neutral egg being added per 100g maize yellow-powders
White enzyme is respectively 1.85-2.95AU and 0.45-0.90AU.
Specifically, maize yellow-powder can be added in retort first, it is proportionally added into deionized water and sizes mixing, and be added
Sodium hydroxide adjusts pH to 8-10；It then is heated to 50-65 DEG C, and alkali protease is added and is digested, enzymolysis is completed
Afterwards, neutral proteinase is continuously added to be digested；Implement destroy the enzyme treatment after the completion of enzymolysis, for example be warming up to 100 DEG C of enzyme deactivations, maintains
30min or so obtains enzymolysis liquid.
For the solid impurity in removal enzymolysis liquid, first enzymolysis liquid can be implemented to centrifuge.The present invention is to centrifugation used
The specific means of separation is not specially limited, for example decanter centrifuge or desk centrifuge can be used and carried out tentatively to enzymolysis liquid
Separating-purifying.
The filtration treatment that enzymolysis liquid after centrifugation is implemented specifically can be used aperture be 50-200nm filter membrane into
Row, is typically chosen the ceramic membrane in the apertures 50-200nm, such as the ceramic membrane in the apertures 100-200nm, further such as the apertures 200nm
Ceramic membrane.After filtering, more clear enzymolysis liquid can be obtained.
Filtered enzymolysis liquid can be handled through activated carbon adsorption first, specifically, in charcoal adsorption process, activated carbon
Mass ratio with maize yellow-powder is (4-25)：100, the grain size of activated carbon is less than 0.0750mm；
The temperature of activated carbon adsorption is 40-60 DEG C, and the time is no less than 10min.
It is appreciated that above-mentioned activated carbon should be food grade active charcoal, such as food-grade wooden powder shaped activated carbon, fineness
100% is sieved by 200 mesh, can be commercially available.
It has passed through the enzymolysis liquid after activated carbon adsorption and cation exchange tree then passed through with the linear flow rate of 1-10mL/min
Fat column starts to collect efflux when ultraviolet detection value reaches 100mAu, stops collecting when ultraviolet detection value is less than 200mAu
Efflux.
Specifically, cation exchange resin column used is the hydrogen type cation exchange resin column pre-equilibrated, and it is used
Cation exchange resin is food-grade cation exchange resin, specially acrylic resin, and particle size is in 0.5-
1.0mm。
Further, to further increase separating-purifying effect, after upper complete sample, continue with deionized water rinse sun from
Sub-exchange resin.
The enzymolysis liquid that have passed through cation exchange resin purifying then passes through anion with the linear flow rate of 1-10mL/min
Exchange resin column starts to collect efflux when ultraviolet detection value reaches 100mAu, stop when ultraviolet detection value is less than 200mAu
Only collect efflux.
Specifically, anion-exchange resin column used is the hydrogen-oxygen type anion-exchange resin column pre-equilibrated, and institute
It is food-grade anion exchange resin, specially acrylic resin with anion exchange resin, particle size is in 0.5-
1.0mm。
Further, to further increase separating-purifying effect, after upper complete sample, continue with deionized water rinse it is cloudy from
Sub-exchange resin.
It is above-mentioned isolate and purify after obtained maize oligopeptide, the arbitrarily acceptable mode in current this field can be used into traveling
One step is processed, and to meet corresponding application demand, for example maize oligopeptide product, Huo Chengwei is obtained after being further dried
Corn oligopeptide powder.Or implement be dried before, concentration is carried out to it first, with shorten drying time and at
This.
Above-mentioned concentration may be used this field Conventional concentration means and complete, for example be concentrated by evaporation.Of the invention specific
It is that the enzymolysis liquid after separating-purifying is concentrated into Baume value using triple effect vacuum concentrator (to use hand for 20% in implementation process
The formula refractometer of holding quickly detects the content of soluble solid in solution, and the numerical value directly read is Baume value, with percentage
Than embodying).
This field conventional drying means can be used in above-mentioned drying process, and the present invention is not specially limited herein, in the present invention
It is that drying is implemented to purified product using spray drying process, spray drying condition is specially in specific implementation process：Import temperature
160-180 DEG C of degree, 60-80 DEG C of outlet temperature.Spray dried products are collected to get to corn oligopeptide powder.
For pEL, VL and the LL for identifying in above-mentioned corn oligopeptide powder, the present invention uses molecular-exclusion chromatography (Superdex
Peptide 10/30GL, GE Healthcare) pre-separation is carried out to maize oligopeptide, recycle reversed-phase high performance liquid chromatography connection
It closes Mass Spectrometric Identification and goes out 3 main feature peptide fragments therein：PEL, VL and LL.
Also, implement process for separating and purifying by using above-mentioned enzymolysis process and cooperation, in obtained maize oligopeptide,
The content of pEL is more than or equal to 0.85%, even up to 1.00% or more；The content of VL is more than or equal to 0.15%, even up to
Content to 0.25% or more, LL is more than or equal to 1.15%, even up to 1.25% or more.
Maize oligopeptide provided by the invention, wherein containing having the different physiological roles such as protect liver, anti-inflammatory and antidepression
PEL peptide fragments, can as VL the and LL peptide fragments of good branched-chain amino acid propylhomoserin replenishers, to be maize oligopeptide product
It provides and is more widely applied foreground.
The industrialized preparing process of maize oligopeptide provided by the invention makes by using specific process for separating and purifying
PEL peptide fragments with different physiological roles such as protect liver, anti-inflammatory and antidepressions and good branched-chain amino acid ammonia can be used as
VL the and LL peptide fragments of sour replenishers are retained, wherein the content of pEL be more than or equal to 0.85%, even up to 1.00% with
On；The content of VL is more than or equal to 0.15%, is even up to more than or equal to 1.15% to the content of 0.25% or more, LL, or even can
Reach 1.25% or more.The loss of functional peptide fragment is not only avoided, and has widened the application range of maize oligopeptide product.
Description of the drawings
Fig. 1 is the standard sample spectrum used in identification pEL in the embodiment of the present invention；
Fig. 2 is the standard sample spectrum used in identification VL in the embodiment of the present invention；
Fig. 3 is the standard sample spectrum used in identification LL in the embodiment of the present invention；
Fig. 4 is pEL content detection collection of illustrative plates in maize oligopeptide obtained in the embodiment of the present invention 1；
Fig. 5 is VL content detection collection of illustrative plates in maize oligopeptide obtained in the embodiment of the present invention 1；
Fig. 6 is LL content detection collection of illustrative plates in maize oligopeptide obtained in the embodiment of the present invention 1；
Fig. 7 is pEL content detection collection of illustrative plates in maize oligopeptide obtained in the embodiment of the present invention 2；
Fig. 8 is VL content detection collection of illustrative plates in maize oligopeptide obtained in the embodiment of the present invention 2；
Fig. 9 is LL content detection collection of illustrative plates in maize oligopeptide obtained in the embodiment of the present invention 2；
Figure 10 is pEL content detection collection of illustrative plates in maize oligopeptide obtained in the embodiment of the present invention 3；
Figure 11 is VL content detection collection of illustrative plates in maize oligopeptide obtained in the embodiment of the present invention 3；
Figure 12 is LL content detection collection of illustrative plates in maize oligopeptide obtained in the embodiment of the present invention 3.
Specific implementation mode
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with the embodiment of the present invention
In attached drawing, technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is
A part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art
The every other embodiment obtained without making creative work, shall fall within the protection scope of the present invention.
In following embodiment and comparative example, utilize reversed-phase high performance liquid chromatography Inertsil ODS-3 (5 μm, 2.1mm ×
250mm, GL Science, Tokyo, Japan) joint mass spectrum (LCMS-8040, Shimadzu, Kyoto, Japan) detection corn
Functional peptide fragment in oligopeptide：The content of pEL, VL and LL.
In following embodiment and comparative example, maize yellow-powder used is purchased from Qinghuangdao Lihua Starch Co., Ltd., and 60
Type maize yellow-powder；Alkali protease is purchased from letter (China) Bioisystech Co., Ltd of Novi, Alcalase 2.4L type alkalinity eggs
White enzyme；Neutral proteinase is purchased from letter (China) Bioisystech Co., Ltd of Novi, Neutrase 0.8L type neutral proteinases.
Embodiment 1
1, the preparation of maize oligopeptide：
1) 500g corn protein powders are added in retort, are sized mixing with 5L deionized waters, NaOH adjustment pH value, which is added, is
9.0,50 DEG C are warming up to, alkali protease 4.5g is added, after digesting 30min, neutral proteinase 3.6g is added, continues to digest
3.5h is warming up to 100 DEG C of enzyme deactivations, maintains 30min.
2) it uses desk centrifuge to carry out centrifugal treating to enzymolysis liquid, collects clear liquid, then use aperture for the ceramics of 200nm
Film is filtered, and further clarifies enzymolysis liquid.
3) addition 20g food-grades wooden powder shaped activated carbon (fineness 100% passes through 200 mesh and sieves) in enzymolysis liquid, 50 DEG C
Lower absorption 30min or so, then removes activated carbon.
4) enzymolysis liquid after activated carbon adsorption is passed through into the hydrogen form cation that has pre-equilibrated with the linear flow rate of 1mL/min
Exchange resin column starts to collect efflux when ultraviolet detection value is higher than 100mAu, continues to spend ionized water punching after upper complete sample
Cation exchange resin column is washed, stops collecting efflux when ultraviolet detection value is less than 200mAu.
5) efflux of collection is passed through into the hydrogen-oxygen type anion exchange resin that has pre-equilibrated with the linear flow rate of 1mL/min
Column starts to collect when ultraviolet detection value is higher than 100mAu, continues to spend ionized water flushing cation exchange tree after upper complete sample
Fat column stops collecting efflux when ultraviolet detection value is less than 200mAu.
6) it concentrates efflux to Baume value 20% with evaporation concentration system to stop, being then fed into spray drying tower, wherein into
160 DEG C of temperature of mouth, 60 DEG C of outlet temperature obtain corn oligopeptide powder product.
2, in maize oligopeptide product functional peptide fragment pEL, VL and LL content detection：
Using molecular-exclusion chromatography (Superdex Peptide 10/30GL, GE Healthcare) to maize oligopeptide
Pre-separation is carried out, using whether containing characteristic peptide fragment in reversed-phase high performance liquid chromatography joint Mass Spectrometric Identification maize oligopeptide：
PEL, VL and LL.
Respectively as shown in Figure 1 to Figure 3 (standard specimen), Fig. 1 includes pEL (m/z=to the Structural Identification collection of illustrative plates of pEL, VL, LL
243.1) first mass spectrometric figure and its secondary fragment collection of illustrative plates under the conditions of CE=-35.0；Fig. 2 includes VL (m/z=231.2)
First mass spectrometric figure and its secondary fragment collection of illustrative plates under the conditions of CE=-35.0；Fig. 3 includes the first mass spectrometric of LL (m/z=245.2)
Figure and its secondary fragment collection of illustrative plates under the conditions of CE=-35.0.
Through being compared with standard specimen (Fig. 1 to Fig. 3), it was demonstrated that exist simultaneously peptide fragment pEL, VL in obtained maize oligopeptide
And LL.Fig. 4 to fig. 6 is the content detection collection of illustrative plates of pEL, VL and LL in maize oligopeptide product made from the present embodiment respectively, warp
Detection, in the maize oligopeptide product, content that the content of functional peptide fragment pEL is 0.88% ± 0.01%, VL is 0.27% ±
The content of 0.00%, LL were 1.20% ± 0.02% (in terms of maize oligopeptide product gross mass, similarly hereinafter).
The method for illustrating to be provided using the present embodiment so that there are the different physiological roles such as protect liver, anti-inflammatory and antidepression
The pEL peptide fragments of property can be retained as VL the and LL peptide fragments of good branched-chain amino acid propylhomoserin replenishers.
Embodiment 2
1, the preparation of maize oligopeptide：
1) 500g corn protein powders are added in retort, are sized mixing with 5L deionized waters, NaOH adjustment pH value, which is added, is
9.0,50 DEG C are warming up to, alkali protease 4.5g is added, after digesting 30min, neutral proteinase 3.6g is added, continues to digest
3.5hr is warming up to 100 DEG C of enzyme deactivations, maintains 30min.
2) it uses desk centrifuge to carry out centrifugal treating to enzymolysis liquid, collects clear liquid, then use aperture for the ceramics of 200nm
Film is filtered, and further clarifies enzymolysis liquid.
3) addition 100g food-grades wooden powder shaped activated carbon (fineness 100% passes through 200 mesh and sieves) in enzymolysis liquid, 50 DEG C
Lower absorption 12min or so, then removes activated carbon.
4) by the enzymolysis liquid after activated carbon adsorption with the linear flow rate of 10mL/min by the Hydrogen sun that has pre-equilibrated from
Sub-exchange resin column starts to collect efflux when ultraviolet detection value is higher than 100mAu, continues to spend ionized water after upper complete sample
Cation exchange resin column is rinsed, stops collecting efflux when ultraviolet detection value is less than 200mAu.
5) efflux of collection is passed through into the hydrogen-oxygen type anion exchange tree that has pre-equilibrated with the linear flow rate of 10mL/min
Fat column starts to collect when ultraviolet detection value is higher than 100mAu, continues to spend ionized water flushing cation exchange after upper complete sample
Resin column stops collecting efflux when ultraviolet detection value is less than 200mAu.
6) it concentrates efflux to Baume value 20% with evaporation concentration system to stop, being then fed into spray drying tower, wherein into
180 DEG C of temperature of mouth, 80 DEG C of outlet temperature obtain corn oligopeptide powder product.
2, in maize oligopeptide product functional peptide fragment pEL, VL and LL content detection：
Method in reference implementation example 1 identifies in the maize oligopeptide product in the present embodiment whether contain pEL, VL and LL
And corresponding content.Fig. 7 to Fig. 9 is the content detection of pEL, VL and LL in maize oligopeptide product made from the present embodiment respectively
Collection of illustrative plates.After testing, peptide fragment pEL, VL and LL are existed simultaneously in the maize oligopeptide, wherein the content of functional peptide fragment pEL is
The content that the content of 0.97% ± 0.01%, VL are 0.17% ± 0.00%, LL is 1.19% ± 0.02%.
The method for illustrating to be provided using the present embodiment so that there are the different physiological roles such as protect liver, anti-inflammatory and antidepression
The pEL peptide fragments of property can be retained as VL the and LL peptide fragments of good branched-chain amino acid propylhomoserin replenishers.
Embodiment 3
1, the preparation of maize oligopeptide：
1) 500g corn protein powders are added in retort, are sized mixing with 5L deionized waters, NaOH adjustment pH value, which is added, is
9.0,50 DEG C are warming up to, alkali protease 4.5g is added, after digesting 30min, neutral proteinase 3.6g is added, continues to digest
3.5hr is warming up to 100 DEG C of enzyme deactivations, maintains 30min.
2) it uses desk centrifuge to carry out centrifugal treating to enzymolysis liquid, collects clear liquid, then use aperture for the ceramics of 200nm
Film is filtered, and further clarifies enzymolysis liquid.
3) addition 50g food-grades wooden powder shaped activated carbon (fineness 100% passes through 200 mesh and sieves) in enzymolysis liquid, 50 DEG C
Lower absorption 30min or so, then removes activated carbon.
4) enzymolysis liquid after activated carbon adsorption is passed through into the hydrogen form cation that has pre-equilibrated with the linear flow rate of 5mL/min
Exchange resin column starts to collect efflux when ultraviolet detection value is higher than 100mAu, continues to spend ionized water punching after upper complete sample
Cation exchange resin column is washed, stops collecting efflux when ultraviolet detection value is less than 200mAu.
5) efflux of collection is passed through into the hydrogen-oxygen type anion exchange resin that has pre-equilibrated with the linear flow rate of 5mL/min
Column starts to collect when ultraviolet detection value is higher than 100mAu, continues to spend ionized water flushing cation exchange tree after upper complete sample
Fat column stops collecting efflux when ultraviolet detection value is less than 200mAu.
6) it concentrates efflux to Baume value 20% with evaporation concentration system to stop, being then fed into spray drying tower, wherein into
170 DEG C of temperature of mouth, 60 DEG C of outlet temperature obtain corn oligopeptide powder product.
2, in corn peptide functional peptide fragment pEL, VL and LL content detection：
Method in reference implementation example 1 identifies in the maize oligopeptide product in the present embodiment whether contain pEL, VL and LL
And corresponding content.Figure 10 to Figure 12 is the content inspection of pEL, VL and LL in maize oligopeptide product made from the present embodiment respectively
Mapping is composed.After testing, peptide fragment pEL, VL and LL are existed simultaneously in the maize oligopeptide, wherein the content of functional peptide fragment pEL is
The content that the content of 1.02% ± 0.01%, VL are 0.24% ± 0.00%, LL is 1.26% ± 0.02%.
The method for illustrating to be provided using the present embodiment so that there are the different physiological roles such as protect liver, anti-inflammatory and antidepression
The pEL peptide fragments of property can be retained as VL the and LL peptide fragments of good branched-chain amino acid propylhomoserin replenishers.
Finally it should be noted that：The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations；To the greatest extent
Present invention has been described in detail with reference to the aforementioned embodiments for pipe, it will be understood by those of ordinary skill in the art that：Its according to
So can with technical scheme described in the above embodiments is modified, either to which part or all technical features into
Row equivalent replacement；And these modifications or replacements, various embodiments of the present invention technology that it does not separate the essence of the corresponding technical solution
The range of scheme.
Claims (10)
1. a kind of maize oligopeptide, which is characterized in that include at least peptide fragment pEL, VL and LL in the maize oligopeptide composition.
2. maize oligopeptide according to claim 1, which is characterized in that the quality based on maize oligopeptide, the pEL's
Content >=1.15% of content >=0.85%, content >=0.15% of VL, LL.
3. maize oligopeptide according to claim 1 or 2, which is characterized in that the maize oligopeptide is by corn
After bloom carries out protease hydrolyzed, then it is obtained through isolating and purifying,
Wherein, described isolate and purify includes implementing centrifugation, filtering, activated carbon adsorption, cation exchange resin to enzymolysis liquid successively
Purifying and anion exchange resin purifying.
4. a kind of industrialized preparing process of claim 1-3 any one of them maize oligopeptides, which is characterized in that including such as
Lower step：
Maize yellow-powder is implemented to digest using protease, obtains enzymolysis liquid；
Implement centrifugation, filtering, activated carbon adsorption, cation exchange resin purifying and anion exchange tree successively to the enzymolysis liquid
Fat purification process.
5. industrialized preparing process according to claim 4, which is characterized in that the enzymolysis step includes：
It after maize yellow-powder is mixed with water, sequentially adds alkali protease and neutral proteinase is digested, and through destroy the enzyme treatment,
The enzymolysis liquid is obtained, wherein：
The mass ratio of maize yellow-powder and water is 1：(5-20)；The alkali protease and neutral proteinase being added per 100g maize yellow-powders
Respectively 1.85-2.95AU and 0.45-0.90AU.
6. industrialized preparing process according to claim 4, which is characterized in that the filtering uses aperture for 50-200nm
Filter membrane carry out.
7. industrialized preparing process according to claim 4, which is characterized in that in charcoal adsorption process, activated carbon
Mass ratio with maize yellow-powder is (4-25)：100, the grain size of the activated carbon is less than 0.0750mm；
The temperature of activated carbon adsorption is 40-60 DEG C, and the time is no less than 10min.
8. industrialized preparing process according to claim 4, which is characterized in that the cation exchange resin purifying is specific
Including：
Enzymolysis liquid after macroporous absorbent resin isolates and purifies is passed through into cation exchange tree with the linear flow rate of 1-10mL/min
Fat column starts to collect efflux when ultraviolet detection value reaches 100mAu, stops collecting when ultraviolet detection value is less than 200mAu
Efflux.
9. industrialized preparing process according to claim 4, which is characterized in that the anion exchange resin purifying is specific
Including：
By the enzymolysis liquid purified through cation exchange resin with the linear flow rate of 1-10mL/min by anion-exchange resin column,
Start to collect efflux when ultraviolet detection value reaches 100mAu, stops collecting outflow when ultraviolet detection value is less than 200mAu
Liquid.
10. industrialized preparing process according to claim 4, which is characterized in that further include implementing to spray to purified product
It is dried, spray drying condition is：160-180 DEG C of inlet temperature, 60-80 DEG C of outlet temperature.
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Cited By (1)
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| WO2021082311A1 (en) * | 2019-10-29 | 2021-05-06 | 中国食品发酵工业研究院有限公司 | Pea peptide having supplementary blood glucose reducing function and preparation method therefor |
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