CN109486889A - A kind of moringa seeds micromolecule polypeptide and its preparation process - Google Patents
A kind of moringa seeds micromolecule polypeptide and its preparation process Download PDFInfo
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Abstract
A kind of moringa seeds micromolecule polypeptide and its preparation process, are related to moringa seeds field of deep.Moringa seeds micromolecule polypeptide mainly uses the moringa seeds dregs of rice and complex enzyme to digest under the conditions of natural pH and obtains, the content of peptides > 50% in moringa seeds micromolecule polypeptide, and polypeptide middle-molecular-weihydroxyethyl is less than peptide fragment content >=85% of 1000D.The preparation process of moringa seeds micromolecule polypeptide is to mix the moringa seeds dregs of rice and water, and heating is homogenized to obtain slurries；Slurries are cooling, under the conditions of natural pH, slurries and the alkali protease for accounting for the neutral proteinase of moringa seeds dregs of rice quality 0.5%-4%, accounting for moringa seeds dregs of rice quality 0.2%-5% are obtained into enzymolysis liquid in 50-60 DEG C of enzymatic hydrolysis 2-8h；It by enzymolysis liquid enzyme deactivation, filters to get filtrate, and filtrate is concentrated, is dry.The content of peptides of moringa seeds micromolecule polypeptide product is higher, and especially micromolecule polypeptide content is high, has good absorbability.
Description
Technical field
The present invention relates to moringa seeds field of deep, and in particular to a kind of moringa seeds micromolecule polypeptide and its preparation work
Skill.
Background technique
Moringa is the plant of Moringaceae Moringa, originates in the Himalayas of north India, be planted in Asia extensively
Continent and African subtropical and tropical zones.Moringa whole body is all precious, blade, flower, seed and tender shoots, tender stem, root etc.
It is edible, it is the plant of integration of drinking and medicinal herbs.Hypertension, diabetes, cardiovascular disease, wind are often treated with Moringa by India and African country
The high-incidence disease such as wet, arthritis.Grease and protein rich in moringa seeds, are good protein sources.Moringa
Seed albumen contains more rich for proton or the amino acid of electron supplying capacity, specifically includes: tyrosinase Tyr, methionine Met,
Cysteine Cys；Also contain more rich hydrophobic amino acid, specifically includes: leucine Leu, proline Pro, phenylalanine
Phe and valine Val；And acidic amino acid abundant: glutamic acid Glu.Therefore, can be prepared using enzyme process is had by force
The moringa seeds peptide of antioxidant activity, moringa seeds peptide can be used as efficient antioxidant for food industry, have stronger society
With economic benefit.
Although the protein content in the product that current enzymatic isolation method extracts moringa seeds may be up to 90%, egg
Content of peptides in white matter is lower, and micromolecule polypeptide content is lower, and micromolecule polypeptide refers to that molecular weight is less than the peptide fragment of 1000D,
Therefore, the absorbent properties of product are poor.
Summary of the invention
In order to solve above-mentioned at least one technical problem of the existing technology, the embodiment of the present invention provides a kind of moringa seeds
The content of peptides of micromolecule polypeptide and its preparation process, product are higher, and especially micromolecule polypeptide content is high, have good absorption
Property.
The embodiment of the present invention solves its technical problem and adopts the following technical solutions to realize.
In a first aspect, providing a kind of moringa seeds micromolecule polypeptide, mainly use the moringa seeds dregs of rice and complex enzyme in nature
Enzymatic hydrolysis obtains under the conditions of pH, the content of peptides > 50% in moringa seeds micromolecule polypeptide, and polypeptide middle-molecular-weihydroxyethyl is less than the peptide of 1000D
Duan Hanliang >=85%.
In above-mentioned technical proposal, moringa seeds micromolecule polypeptide is using the moringa seeds dregs of rice and complex enzyme enzyme under the conditions of natural pH
Solution obtains, and realizes the higher value application of the moringa seeds dregs of rice, avoids the waste of moringa seeds albumen, while nature pH enzymolysis process saves
Artificial the step of adjusting pH, reduce work hours, reduces production cost, avoid ash impurities from bringing product into, be conducive to product quality
Raising.The content of peptides > 50% of moringa seeds micromolecule polypeptide, i.e. content of peptides are high, and especially polypeptide middle-molecular-weihydroxyethyl is less than
Peptide fragment content >=85% of 1000D, i.e. micromolecule polypeptide content are high, and small molecule peptide fragment has superior absorbability and low sensitization
Property, therefore, moringa seeds micromolecule polypeptide has good absorbent properties.
In one possible implementation, 10% aqueous solution of mass concentration that moringa seeds micromolecule polypeptide is prepared exists
Light transmittance >=80% of 620nm.
In above-mentioned technical proposal, moringa seeds micromolecule polypeptide prepare 10% aqueous solution of mass concentration 620nm light transmission
Rate reaches 80% or more, and the higher dissolubility for showing product of light transmittance is higher, therefore the dissolubility of product is good, has a wide range of application
It is general, for example the dosage forms such as pulvis, oral solution can be made；And the absorbent properties of product are good.
In one possible implementation, complex enzyme include the neutral proteinase for accounting for moringa seeds dregs of rice quality 0.5%-4%,
Account for moringa seeds dregs of rice quality 0.2%-5% alkali protease.
In above-mentioned technical proposal, under the conditions of natural pH, carried out using a certain amount of neutral proteinase and alkali protease
Enzymatic hydrolysis, can obtain relatively optimal hydrolysis result.
In one possible implementation, complex enzyme further includes the zytase for accounting for moringa seeds dregs of rice quality 0.5%-2%,
And/or account for the 1,4 beta-glucanase of moringa seeds dregs of rice quality 0.2%-1%.
In above-mentioned technical proposal, under the conditions of natural pH, neutral proteinase and alkali protease can utilize system itself
PH and its variation sufficiently digest the moringa seeds dregs of rice with a certain amount of zytase collective effect；Under the conditions of natural pH,
Neutral proteinase and alkali protease can utilize pH and its variation of system itself, make jointly with a certain amount of 1,4 beta-glucanase
With sufficiently being digested to the moringa seeds dregs of rice.
Second aspect provides a kind of preparation process of moringa seeds micromolecule polypeptide comprising following steps:
The moringa seeds dregs of rice and water 1:10-20 in mass ratio are mixed, are heated to 90-105 DEG C, 0.5-2h is homogenized, obtains slurries；
Slurries are cooled to 50-60 DEG C, under the conditions of natural pH, by slurries and account for moringa seeds dregs of rice quality 0.5%-4%'s
Neutral proteinase, the alkali protease for accounting for moringa seeds dregs of rice quality 0.2%-5% obtain enzymolysis liquid in 50-60 DEG C of enzymatic hydrolysis 2-8h；
It by enzymolysis liquid enzyme deactivation, filters to get filtrate, and filtrate is concentrated, is dry.
In above-mentioned technical proposal, using the moringa seeds dregs of rice as raw material, it is able to achieve the higher value application of the moringa seeds dregs of rice, is avoided
The waste of moringa seeds albumen；The moringa seeds dregs of rice and water are homogenized in a certain temperature conditions, it can will be effective in the moringa seeds dregs of rice
Ingredient is dissolved in the water, and does not destroy the activity of effective component, to carry out subsequent enzymolysis step；In natural pH condition
Under, it is digested in a certain temperature conditions using a certain amount of neutral proteinase and alkali protease, saves artificial adjusting
The step of pH, reduces work hours, and reduces production cost, avoids ash impurities from bringing product into, be conducive to the raising of product quality.
In one possible implementation, the method for enzyme deactivation are as follows: in 95-100 DEG C of holding 10-15min；And/or vacuum
The vacuum degree used is concentrated as 0.06-0.08.
In above-mentioned technical proposal, enzyme deactivation is carried out using certain high temperature and time-triggered protocol, relatively minimal energy can be consumed
Inactivate the complex enzymes such as neutral proteinase and alkali protease；It is concentrated using certain vacuum degree, can quickly go to remove water
Point, realize concentration purpose.
In one possible implementation, the condition of spray drying are as follows: inlet air temperature is 180-220 DEG C, leaving air temp
It is 60-95 DEG C, air inducing rotor power is 30-40hz, spray head rotor power 15-20hz.
It in above-mentioned technical proposal, is spray-dried using certain condition, epigranular can be obtained, and main component is
The solid product of moringa seeds polypeptide, convenient for storage and application.
In one possible implementation, alkali protease is made according to following preparation method: by Svf4-21-7 bacterium
Kind expands culture through seed step by step, is transferred to ferment tank, obtains alkali protease fermentation liquid；By alkali protease fermentation liquid through sulphur
It is freeze-dried after acid ammonium salt analysis, dialysis desalting, ion-exchange chromatography, polyethylene glycol embedding concentration, gel permeation chromatography, alkali is made
Property protease solid powder；
And/or neutral proteinase is made according to following preparation method: AS1.398 strain is expanded training through seed step by step
It supports, is transferred to ferment tank, obtains seed liquor；Flocculant is added in seed liquor, obtains filtrate；By filtrate centrifugation, coarse filtration, refined filtration,
Trehalose and cycloheptaamylose are added, precipitating is collected in stirring, and neutral proteinase solid powder is made in vacuum drying.
It is high according to alkali protease made from above-mentioned preparation method and neutral proteinase activity in above-mentioned technical proposal, hold
The activity of controlled enzymatic hydrolysis step easily in technique production.
In one possible implementation, it is additionally added the zytase progress enzyme for accounting for moringa seeds dregs of rice quality 0.5%-2%
Solution, zytase are made according to following preparation method: the slant strains of bacillus alcalophilus CCTCC NO:M2013537 are passed through
Activation, multistage seed culture and first class seed pot expand culture step by step and obtain seed liquor；Seed liquor is connect by 5%-8% inoculum concentration
Enter fermentor, in 37-45 DEG C of constant temperature incubation 10-15h；It is cooled to 10-15 DEG C, constant temperature incubation 15-20h；Continue to be cooled to 2-5
DEG C, by seed liquor with the additional access fermentor of 2%-5% inoculum concentration, constant temperature incubation 20-30h；It is warming up to 10-15 DEG C, constant temperature training
Support 15-20h；It is continuously heating to 37-45 DEG C, constant temperature incubation 15-20h obtains fermentation liquid；By fermentation liquid through filtering, concentration, refined filtration,
Dry zytase solid powder.
In above-mentioned technical proposal, according to the height of xylanase activity made from above-mentioned preparation method, under the conditions of natural pH, in
Property protease and alkali protease can using system itself pH and its variation, it is right with a certain amount of zytase collective effect
The moringa seeds dregs of rice are sufficiently digested.
In one possible implementation, it is additionally added the 1,4 beta-glucanase progress for accounting for moringa seeds dregs of rice quality 0.2%-1%
Enzymatic hydrolysis, 1,4 beta-glucanase are made according to following preparation method: by the inclined-plane bacterium of bacillus licheniformis CCTCC NO:M2013538
The activated culture of kind obtains seed liquor；Seed liquor is accessed in fermentor with 2%-5% inoculum concentration, in 35-45 DEG C of constant temperature incubation
10-15h；Then fermentation medium is filled into the fermenter, in 35-45 DEG C of constant temperature incubation 15-20h；By seed liquor with 1%-4%
The additional access fermentor of inoculum concentration, constant temperature incubation 10-15h；Continuation fills into fermentation medium in the fermenter, in 35-45 DEG C of perseverance
Temperature culture 10-15h, obtains fermentation liquid；By fermentation liquid through filtering, concentration, refined filtration, dry 1,4 beta-glucanase solid powder.
In above-mentioned technical proposal, according to the height of 1,4 beta-glucanase activity made from above-mentioned preparation method, under the conditions of natural pH,
Neutral proteinase and alkali protease can utilize pH and its variation of system itself, make jointly with a certain amount of 1,4 beta-glucanase
With sufficiently being digested to the moringa seeds dregs of rice.
Detailed description of the invention
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is the protein and peptide absorptivity result figure of different product in the embodiment of the present invention.
Specific embodiment
It in order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below will be in the embodiment of the present invention
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, according to normal conditions or manufacturer builds
The condition of view carries out.Reagents or instruments used without specified manufacturer is the conventional production that can be obtained by commercially available purchase
Product.
The moringa seeds micromolecule polypeptide and its preparation process of the embodiment of the present invention are specifically described below.
The embodiment of the present invention provides a kind of moringa seeds micromolecule polypeptide, mainly uses the moringa seeds dregs of rice and complex enzyme certainly
It digests and obtains under the conditions of right pH, the content of peptides > 50% in moringa seeds micromolecule polypeptide, polypeptide middle-molecular-weihydroxyethyl is less than 1000D's
Peptide fragment content >=85%；Optionally, light transmittance of 10% aqueous solution of mass concentration that moringa seeds micromolecule polypeptide is prepared in 620nm
>=80%.
In the present embodiment, complex enzyme may include the neutral proteinase for accounting for moringa seeds dregs of rice quality 0.5%-4%, account for moringa seeds
Dregs of rice quality 0.2%-5% alkali protease.In other embodiments, complex enzyme can also include accounting for moringa seeds dregs of rice quality 1%-
3% neutral proteinase accounts for moringa seeds dregs of rice quality 1%-3% alkali protease.For example, complex enzyme can also include accounting for moringa seeds
The neutral proteinase of dregs of rice quality 0.5%, 1%, 2%, 3%, 4%, account for moringa seeds dregs of rice quality 0.2%, 1%, 2%, 3%, 4%,
5% alkali protease.
In the present embodiment, complex enzyme further includes the zytase for accounting for moringa seeds dregs of rice quality 0.5%-2%, for example, complex enzyme
It further include the zytase for accounting for moringa seeds dregs of rice quality 0.5%, 1%, 1.5%, 2%；And/or complex enzyme further includes accounting for moringa seeds
The 1,4 beta-glucanase of dregs of rice quality 0.2%-1%, for example, complex enzyme further includes accounting for moringa seeds dregs of rice quality 0.2%, 0.5%, 1%
1,4 beta-glucanase.
The embodiment of the present invention provides a kind of preparation process of moringa seeds micromolecule polypeptide comprising following steps:
(1), the moringa seeds dregs of rice and water 1:10-20 in mass ratio are mixed, is heated to 90-105 DEG C, be homogenized 0.5-2h, must starch
Liquid.
In other embodiments, the moringa seeds dregs of rice and water can also 1:12-18 in mass ratio, such as 1:10,1:12,1:15,
1:18,1:20 mixing；In other embodiments, 95-100 DEG C can also be heated to, for example, be heated to 90 DEG C, 95 DEG C, 100 DEG C,
105℃；In other embodiments, 1-1.5h, such as homogenate 0.5h, 1h, 1.5h, 2h can also be homogenized.
(2), slurries are cooled to 50-60 DEG C, under the conditions of natural pH, by slurries and account for moringa seeds dregs of rice quality 0.5%-
4% neutral proteinase, the alkali protease for accounting for moringa seeds dregs of rice quality 0.2%-5% obtain enzyme in 50-60 DEG C of enzymatic hydrolysis 2-8h
Solve liquid.
It in other embodiments, can also be in 52-57 DEG C of enzymatic hydrolysis 4-6h, for example, in 50 DEG C, 52 DEG C, 55 DEG C, 57 DEG C, 60
DEG C enzymatic hydrolysis 2h, 4h, 6h, 8h.
(3), by enzymolysis liquid enzyme deactivation, filter to get filtrate, and by filtrate concentration, dry.
In the present embodiment, the method for enzyme deactivation are as follows: in 95-100 DEG C of holding 10-15min, for example, in 95 DEG C, 98 DEG C, 100 DEG C
Enzyme deactivation 10min, 12min, 15min.And/or the vacuum degree used is concentrated in vacuo for 0.06-0.08, for example, vacuum degree is
0.06、0.065、0.07、0.08。
In the present embodiment, the condition of spray drying are as follows: inlet air temperature is 180-220 DEG C, such as 180 DEG C, 185 DEG C, 190
DEG C, 200 DEG C, 210 DEG C, 220 DEG C, leaving air temp is 60-95 DEG C, such as 60 DEG C, 65 DEG C, 70 DEG C, 75 DEG C, 80 DEG C, 85 DEG C, 87
℃,90℃,95℃；Air inducing rotor power is 30-40hz, such as 30,35,40hz, spray head rotor power 15-20hz, such as 15,
18、19hz。
Feature and performance of the invention are described in further detail with reference to embodiments.
A, it prepares neutral proteinase: bacillus subtilis AS1.398 strain being expanded into culture through seed step by step, is transferred to fermentation
Tank fermentation, obtains seed liquor；Flocculant is added in seed liquor, is stirred evenly, and is stood 30 minutes, is combined into impurity with flocculant
Seed liquor after the completion of flocculation reaction is carried out plate-frame filtering, obtains filtrate by bulk；By filtrate high speed centrifugation, revolving speed is
10000rpm, feed flow rate 80L/h collect centrifugate；Coarse filtration uses aperture for the Kynoar of 0.22um and 0.1um
Filter membrane is filtered, operating pressure 0.1MPa；It uses molecular cut off to be filtered concentration for the polysulfone ultrafiltration membrane of 10K, operates
Pressure 0.1MPa, obtains concentrate；Enzyme activity protective agent trehalose and cycloheptaamylose are added, the weight of trehalose is the weight of concentrate
The 1% of amount, the weight of cycloheptaamylose are the 0.5% of the weight of concentrate, are stirred evenly；Edible second is added when being slowly stirred
Alcohol, collects cotton-shaped neutral proteinase precipitating, and neutral proteinase solid powder is made in vacuum drying.
B, alkali protease is prepared: by alkali protease superior strain Svf4-21-7 (CGMCC No.6417) strain through planting
Son expands culture step by step, is transferred to ferment tank, obtains alkali protease fermentation liquid；By alkali protease fermentation liquid through ammonium sulfate
It is freeze-dried after analysis, dialysis desalting, ion-exchange chromatography, polyethylene glycol embedding concentration, gel permeation chromatography, basic protein is made
Enzyme solid powder.
C, it prepares zytase: the slant strains of intact bacillus alcalophilus CCTCC NO M2013537 is passed through
Activation and one, two, three seed culture and first class seed pot expand culture step by step and obtain seed liquor, are accessed and are sent out with 6% inoculum concentration
Fermentation tank, cultivates 10-15h by 40 DEG C of cultivation temperature, mixing speed 500rpm, ventilation quantity 1:2V/V；Then with 2 DEG C/h rate of temperature fall
Slow cooling is to 10 DEG C, constant temperature incubation 20h；Continue with 2 DEG C/h rate of temperature fall slow cooling to 4 DEG C, at this point, by seed liquor with
The additional access fermentor of 4% inoculum concentration, constant temperature incubation is for 24 hours；10 DEG C finally are to slowly warm up to 2 DEG C/h heating rate, constant temperature training
Support 15h；Continue to be to slowly warm up to 40 DEG C with 2 DEG C/h heating rate, constant temperature incubation 20h obtains fermentation liquid；Fermentation liquid is filtered, is dense
Contracting, refined filtration, dry zytase solid powder.
D, prepare 1,4 beta-glucanase: the slant strains activation culture of bacillus licheniformis CCTCC NO:M2013538 obtains
Seed liquor；By seed liquor in fermentor of the 3% inoculum concentration access containing 3L fermentation medium, 40 DEG C of cultivation temperature, stirring is fast
Spend 300r/min, ventilation quantity 1:2V/V, incubation time 10, h；It then is 10 DEG C by 121 DEG C of sterilizing 20min, temperature by 1L
Fermentation medium fills into fermentor, constant temperature incubation 20h when temperature rises to 40 DEG C；At this point, seed liquor is additional with 2% inoculum concentration
Access fermentor, constant temperature incubation 12h；Continue to fill into 1L by 121 DEG C of sterilizing 20min, the fermentation medium that temperature is 10 DEG C
Fermentor, constant temperature incubation 12h when temperature rises to 40 DEG C, obtains fermentation liquid；By fermentation liquid through filtering, concentration, refined filtration, dry β-
Dextranase solid powder.
Embodiment 1
The present embodiment provides a kind of moringa seeds micromolecule polypeptides, are made according to following preparation process:
Step 1, the moringa seeds dregs of rice and water 1:15 in mass ratio are heated to 100 DEG C, are homogenized 1h, obtain slurries.
Slurries are cooled to 55 DEG C by step 2, under the conditions of natural pH, are added in slurries and are accounted for moringa seeds dregs of rice quality 3%
Neutral proteinase solid powder, the alkali protease solid powder for accounting for moringa seeds dregs of rice quality 3% digest 6h, obtain enzymolysis liquid.
Step 3, by enzymolysis liquid in 100 DEG C of enzyme deactivation 15min, filter to get filtrate.
Filtrate is concentrated in vacuo by step 4 in vacuum degree 0.065.
Step 5, spray drying, control each technological parameter are as follows: and 185 DEG C of inlet air temperature, 87 DEG C of leaving air temp, air inducing rotor function
Moringa seeds micromolecule polypeptide is made in rate 35hz, spray head rotor power 18hz.
Through detecting, content of peptides is 55% in the moringa seeds micromolecule polypeptide of the present embodiment, and molecular weight is less than the peptide of 1000D
Duan Hanliang is up to 90% or more, and the T60 of 10% aqueous solution is 82%.
Embodiment 2
The present embodiment provides a kind of moringa seeds micromolecule polypeptides, are made according to following preparation process:
Step 1, the moringa seeds dregs of rice are mixed with water 1:10 in mass ratio, are heated to 90 DEG C, are homogenized 2h, are obtained slurries.
Slurries are cooled to 50 DEG C by step 2, under the conditions of natural pH, are added in slurries and are accounted for moringa seeds dregs of rice quality 4%
Neutral proteinase solid powder, the alkali protease solid powder for accounting for moringa seeds dregs of rice quality 1% digest 6h, obtain enzymolysis liquid.
Step 3, by enzymolysis liquid in 100 DEG C of enzyme deactivation 10min, filter to get filtrate.
Filtrate is concentrated in vacuo by step 4 in vacuum degree 0.06.
Step 5, spray drying, control each technological parameter are as follows: and 180 DEG C of inlet air temperature, 70 DEG C of leaving air temp, air inducing rotor function
Moringa seeds micromolecule polypeptide is made in rate 30hz, spray head rotor power 15hz.
Through detecting, content of peptides is 53% in the moringa seeds micromolecule polypeptide of the present embodiment, and molecular weight is less than the peptide of 1000D
Duan Hanliang is up to 87% or more, and the T60 of 10% aqueous solution is 81%.
Embodiment 3
The present embodiment provides a kind of moringa seeds micromolecule polypeptides, are made according to following preparation process:
Step 1, the moringa seeds dregs of rice are mixed with water 1:20 in mass ratio, are heated to 100 DEG C, are homogenized 0.5h, are obtained slurries.
Slurries are cooled to 60 DEG C by step 2, under the conditions of natural pH, are added in slurries and are accounted for moringa seeds dregs of rice quality 1%
Neutral proteinase solid powder, the alkali protease solid powder for accounting for moringa seeds dregs of rice quality 4% digest 6h, obtain enzymolysis liquid.
Step 3, by enzymolysis liquid in 100 DEG C of enzyme deactivation 10min, filter to get filtrate.
Filtrate is concentrated in vacuo by step 4 in vacuum degree 0.08.
Step 5, spray drying, control each technological parameter are as follows: and 190 DEG C of inlet air temperature, 95 DEG C of leaving air temp, air inducing rotor function
Moringa seeds micromolecule polypeptide is made in rate 40hz, spray head rotor power 20hz.
Through detecting, content of peptides is 51% in the moringa seeds micromolecule polypeptide of the present embodiment, and molecular weight is less than the peptide of 1000D
Duan Hanliang is up to 88% or more, and the T60 of 10% aqueous solution is 85%.
Embodiment 4
The present embodiment provides a kind of moringa seeds micromolecule polypeptide, the preparation process substantially phases of preparation process and embodiment 1
Together, the difference is that:
Under the conditions of natural pH, in slurries be added account for moringa seeds dregs of rice quality 2% neutral proteinase solid powder, account for it is peppery
The alkali protease solid powder of the wooden seed dregs of rice quality 2%, the zytase solid powder for accounting for moringa seeds dregs of rice quality 2%, finally make
Obtain moringa seeds micromolecule polypeptide.
Through detecting, content of peptides is 57% in the moringa seeds micromolecule polypeptide of the present embodiment, and molecular weight is less than the peptide of 1000D
Duan Hanliang is up to 91% or more, and the T60 of 10% aqueous solution is 83%.
Embodiment 5
The present embodiment provides a kind of moringa seeds micromolecule polypeptide, the preparation process substantially phases of preparation process and embodiment 1
Together, the difference is that:
Under the conditions of natural pH, in slurries be added account for moringa seeds dregs of rice quality 2% neutral proteinase solid powder, account for it is peppery
The alkali protease solid powder of the wooden seed dregs of rice quality 2%, the 1,4 beta-glucanase solid powder for accounting for moringa seeds dregs of rice quality 2%, finally
Moringa seeds micromolecule polypeptide is made.
Through detecting, content of peptides is 58% in the moringa seeds micromolecule polypeptide of the present embodiment, and molecular weight is less than the peptide of 1000D
Duan Hanliang is up to 93% or more, and the T60 of 10% aqueous solution is 83%.
Embodiment 6
The present embodiment provides a kind of moringa seeds micromolecule polypeptide, the preparation process substantially phases of preparation process and embodiment 1
Together, the difference is that:
Under the conditions of natural pH, in slurries be added account for moringa seeds dregs of rice quality 2% neutral proteinase solid powder, account for it is peppery
The alkali protease solid powder of the wooden seed dregs of rice quality 2% accounts for 1% zytase solid powder of moringa seeds dregs of rice quality, 1% β-
Moringa seeds micromolecule polypeptide is finally made in dextranase solid powder.
Through detecting, content of peptides is 60% in the moringa seeds micromolecule polypeptide of the present embodiment, and molecular weight is less than the peptide of 1000D
Duan Hanliang is up to 94% or more, and the T60 of 10% aqueous solution is 87%.
Comparative example 1
This comparative example provides a kind of method of moringa seeds protein peptides, extracting method the following steps are included:
(1) thermophilic digestion: decladding after the drying of fresh moringa seeds is crushed, 60 meshes, boiling Moringa under the conditions of 100 DEG C are crossed
Seed powder 30min.
(2) high-pressure homogeneous: deionized water is added according to solid-liquid ratio 1:6g/mL in the moringa seeds powder after thermophilic digestion of learning from else's experience, with
The rate of 8000rpm is sheared 10 minutes, is crossed colloid mill 2 times, obtains moringa seeds homogenate, and 20MPa high-pressure homogeneous 2 times.
(3) alkalinity extraction: being added sodium hydroxide in moringa seeds powder homogenate, adjusts pH value to 7.0, is 40 DEG C in temperature
Under conditions of, 40min is extracted, is cooled to room temperature, obtains suspension.
(4) digest: adjusting suspension pH value to 7.0, be added protease that quality is moringa seeds quality 1% (pancreatin and
The mass ratio of NS37071 protease is 2:3), it is digested under conditions of 50 DEG C 18 hours, 90 DEG C of enzyme deactivation 30min, is centrifuged 20min；
Separation, removes upper layer grease, and gained supernatant is enzymolysis liquid.
(5) be classified alcohol precipitation: vacuum concentration enzymolysis liquid to solid content 30% is added the pre- dehydrated alcohol for being cooled to 4 DEG C, makes
In system ethyl alcohol mass content reach after 10%, 200r/min room temperature at the uniform velocity stir 4h, centrifugation 20min obtains supernatant；It will
Supernatant is concentrated into solid content 30%, the pre- dehydrated alcohol for being cooled to 4 DEG C is added, so that ethyl alcohol mass content reaches in system
After 40%, 200r/min room temperature at the uniform velocity stir 4h, it is centrifuged 20min, is precipitated；The deionized water of 8 times of weight in wet base of precipitating is added, it is molten
Xie Hou is concentrated in vacuo, and freeze-drying obtains powder；
(6) macroporous resin enrichment: powder is soluble in water, concentration 100mg/mL crosses Amberlite XAD-16 macropore tree
Rouge column, using 0-60vol% ethanol solution carry out 8 column volume gradient elutions, flow velocity 2mL/min, specifically: 1-5 column
Volume uses 100vol% water, 0vol% ethanol elution；6-7 column volume uses 50vol% water, 50vol% ethanol elution；7-
8 column volumes use 40vol% water, 60vol% ethanol elution；60% (v/v) ethanol eluate is collected, freeze-drying must have
There are the moringa seeds protein peptides of strong anti-oxidative activity.
It is 92% through protein content in detection moringa seeds protein peptides, moisture content 7%, ORAC value is 1455 μm of ol
Trolox equiv/g。
Below by way of isolated rat test for intestinal absorption to the absorbability of the moringa seeds micromolecule polypeptide of the embodiment of the present invention
It can be carried out evaluation.
Weight 180-200gSD rat is selected in as animal subject, barbital sodium solution is injected into rat abdominal cavity and carries out fiber crops
It is liquor-saturated, open abdomen, by stomachus pyloricus position confirm duodenum, jejunum upper end is ligatured, in ligation port side cut small intestine, and by its
7CM is removed, is placed in Tyrode liquid and cleans 3-4 times, and is overturn in ligation mouth side insertion glass bar, removal enteral is held
Object, for use.
The small intestine broken ends of fractured bone is inserted at the tip for the horminess glass tube that both ends are open, and is tightened the small intestine broken ends of fractured bone with cotton thread, by
The butt end of horminess glass tube injects test liquid 10ml to the Tyrode liquid of intestinal tube serosa side injection 2ml in another Boiling tube,
As mucous membrane side liquid.Specifically provide three kinds of test liquids tested, respectively moringa seeds cypress prepare solution, embodiment 1 it is peppery
Solution, the soya-bean polypeptides (purchase is in Wuhan Tiantianhao Biology Products Co., Ltd) of the wooden seed micromolecule polypeptide preparation are prepared molten
Liquid, test liquid are to be formulated with Tyrode liquid, and mass concentration is 0.1%.
It puts it into 37 DEG C of thermostats to fix, be continually fed into mixed gas (+95% oxygen of 5% carbon dioxide), cultivate 1h,
The liquid extracted in mucous membrane side liquid and horminess glass tube is detected, and measures liquid protein content of peptides using Forint phenol method,
Shown in its result figure 1.
As seen from Figure 1, moringa seeds micromolecule polypeptide is substantially better than the moringa seeds dregs of rice in small intestine in the absorptivity of small intestine
Absorptivity, and difference has extremely significant property；Although absorptivity of the moringa seeds micromolecule polypeptide in small intestine is slightly below soya-bean polypeptides
Absorptivity in small intestine, but do not have otherness, the comprehensive moringa seeds micromolecule polypeptide for illustrating the embodiment of the present invention has excellent
Absorbent properties more.
In conclusion the moringa seeds micromolecule polypeptide and its preparation process of the embodiment of the present invention, the content of peptides of product compared with
Height, especially micromolecule polypeptide content are high, have good absorbability.
Embodiments described above is a part of the embodiment of the present invention, instead of all the embodiments.Reality of the invention
The detailed description for applying example is not intended to limit the range of claimed invention, but is merely representative of selected implementation of the invention
Example.Based on the embodiments of the present invention, obtained by those of ordinary skill in the art without making creative efforts
Every other embodiment, shall fall within the protection scope of the present invention.
Claims (10)
1. a kind of moringa seeds micromolecule polypeptide, which is characterized in that it mainly uses the moringa seeds dregs of rice and complex enzyme in natural pH item
Enzymatic hydrolysis obtains under part, the content of peptides > 50% in the moringa seeds micromolecule polypeptide, and the polypeptide middle-molecular-weihydroxyethyl is less than 1000D
Peptide fragment content >=85%.
2. moringa seeds micromolecule polypeptide according to claim 1, which is characterized in that the moringa seeds micromolecule polypeptide is prepared
10% aqueous solution of mass concentration 620nm light transmittance >=80%.
3. moringa seeds micromolecule polypeptide according to claim 1, which is characterized in that the complex enzyme includes accounting for the Moringa
Neutral proteinase, the Zhan Suoshu moringa seeds dregs of rice quality 0.2%-5% alkali protease of seed dregs of rice quality 0.5%-4%.
4. moringa seeds micromolecule polypeptide according to claim 3, which is characterized in that the complex enzyme further include account for it is described peppery
The zytase of the wooden seed dregs of rice quality 0.5%-2%, and/or, Zhan Suoshu moringa seeds dregs of rice quality 0.2%-1% 1,4 beta-glucanase.
5. a kind of preparation process of moringa seeds micromolecule polypeptide, which is characterized in that itself the following steps are included:
The moringa seeds dregs of rice and water 1:10-20 in mass ratio are mixed, are heated to 90-105 DEG C, 0.5-2h is homogenized, obtains slurries；
The slurries are cooled to 50-60 DEG C, under the conditions of natural pH, by the slurries and account for the moringa seeds dregs of rice quality
The alkali protease of the neutral proteinase of 0.5%-4%, Zhan Suoshu moringa seeds dregs of rice quality 0.2%-5%, in 50-60 DEG C of enzymatic hydrolysis 2-
8h obtains enzymolysis liquid；
It by the enzymolysis liquid enzyme deactivation, filters to get filtrate, and the filtrate is concentrated, is dry.
6. the preparation process of moringa seeds micromolecule polypeptide according to claim 5, which is characterized in that the method for the enzyme deactivation
Are as follows: in 95-100 DEG C of holding 10-15min；And/or the vacuum degree used is concentrated in vacuo as 0.06-0.08.
7. the preparation process of moringa seeds micromolecule polypeptide according to claim 5, which is characterized in that the condition of spray drying
Are as follows: inlet air temperature is 180-220 DEG C, and leaving air temp is 60-95 DEG C, and air inducing rotor power is 30-40hz, spray head rotor power
15-20hz。
8. the preparation process of moringa seeds micromolecule polypeptide according to claim 5, which is characterized in that the alkali protease
It is to be made according to following preparation method: Svf4-21-7 strain is expanded into culture through seed step by step, ferment tank is transferred to, obtains alkali
Property protease fermented liquid；By alkali protease fermentation liquid through ammonium sulfate precipitation, dialysis desalting, ion-exchange chromatography, polyethylene glycol
It is freeze-dried after embedding concentration, gel permeation chromatography, alkali protease solid powder is made；
And/or the neutral proteinase is made according to following preparation method: AS1.398 strain is expanded training through seed step by step
It supports, is transferred to ferment tank, obtains seed liquor；Flocculant is added in the seed liquor, obtains filtrate；By filtrate centrifugation, coarse filtration, essence
Trehalose and cycloheptaamylose are added in filter, and precipitating is collected in stirring, and neutral proteinase solid powder is made in vacuum drying.
9. the preparation process of moringa seeds micromolecule polypeptide according to claim 5, which is characterized in that be additionally added account for it is described peppery
The zytase of the wooden seed dregs of rice quality 0.5%-2% is digested, and the zytase is made according to following preparation method: will be thermophilic
The slant strains of hot bacillus CCTCC NO:M2013537 are activated, multistage seed culture and first class seed pot expand step by step
Culture obtains seed liquor；The seed liquor is accessed into fermentor by 5%-8% inoculum concentration, in 37-45 DEG C of constant temperature incubation 10-15h；
It is cooled to 10-15 DEG C, constant temperature incubation 15-20h；Continue to be cooled to 2-5 DEG C, the seed liquor is additional with 2%-5% inoculum concentration
Access fermentor, constant temperature incubation 20-30h；It is warming up to 10-15 DEG C, constant temperature incubation 15-20h；It is continuously heating to 37-45 DEG C, constant temperature
15-20h is cultivated, fermentation liquid is obtained；By the fermentation liquid through filtering, concentration, refined filtration, dry zytase solid powder.
10. the preparation process of moringa seeds micromolecule polypeptide according to claim 5, which is characterized in that be additionally added described in accounting for
The 1,4 beta-glucanase of moringa seeds dregs of rice quality 0.2%-1% is digested, and 1,4 beta-glucanase is made according to following preparation method: will
The activated culture of the slant strains of bacillus licheniformis CCTCC NO:M2013538 obtains seed liquor；By the seed liquor with
2%-5% inoculum concentration accesses in fermentor, in 35-45 DEG C of constant temperature incubation 10-15h；Then fermented and cultured is filled into the fermenter
Base, in 35-45 DEG C of constant temperature incubation 15-20h；By the seed liquor with the additional access fermentor of 1%-4% inoculum concentration, constant temperature incubation
10-15h；Continuation fills into fermentation medium in the fermenter, in 35-45 DEG C of constant temperature incubation 10-15h, obtains fermentation liquid；By the hair
Zymotic fluid is through filtering, concentration, refined filtration, dry 1,4 beta-glucanase solid powder.
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