CN108753885B - The preparation method and queen bee nit glucosidase suppressor activity peptide of a kind of queen bee nit glucosidase suppressor activity peptide and application - Google Patents
The preparation method and queen bee nit glucosidase suppressor activity peptide of a kind of queen bee nit glucosidase suppressor activity peptide and application Download PDFInfo
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- CN108753885B CN108753885B CN201810597298.7A CN201810597298A CN108753885B CN 108753885 B CN108753885 B CN 108753885B CN 201810597298 A CN201810597298 A CN 201810597298A CN 108753885 B CN108753885 B CN 108753885B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
Abstract
The present invention provides a kind of preparation methods of queen bee nit glucosidase suppressor activity peptide, belong to functional food technical field, including queen bee larva successively through enzymatic hydrolysis, centrifuge separation, filtering and ultrafiltration step.Preparation method provided by the invention can prepare molecular weight be [3~5) active peptide of kDa, be able to suppress glucuroide.
Description
Technical field
The invention belongs to functional food technical fields, and in particular to a kind of queen bee nit glucosidase suppressor activity peptide
Preparation method and queen bee nit glucosidase suppressor activity peptide and application.
Background technique
Queen bee nit is honeybee fertilized eggs on the royal cell not covered, through worker bee feeding royal jelly after hatching to before nymphosis
Larva body, be the byproduct during royal jelly and its product.Queen bee nit is very nutritious, and protein content is high, has very
High edible and medical value, but queen bee nit resource domestic at present is only simply processed, or will be directly as raw material
Outlet, the technology content and added value of product are low, and the resource of most queen bee nits is not developed and used effectively yet.
Correlative study shows that hydrolysis final product of the protein in digestion under albumen enzyme effect is mostly 2~4
The small peptide of a amino acid residue composition.Because small peptide structure is simple, molecular weight is small, unsaturated, so can pass through the infiltration of cell membrane
It is directly entered into the cell with original shape thoroughly, without re-digesting, does not also need to expend energy.Therefore the absorption of small-molecular peptides,
Conversion and utilization, are all efficient, complete, thorough.
Summary of the invention
In view of this, the purpose of the present invention is to provide a kind of preparation sides of queen bee nit glucosidase suppressor activity peptide
Method can obtain queen bee nit glucosidase suppressor activity peptide, which is able to suppress glucosidase activity.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention provides a kind of preparation methods of queen bee nit glucosidase suppressor activity peptide, comprising the following steps:
1) queen bee larva is mixed into homogenate with water, after the pH value for adjusting homogenate is 6~7, is mixed with protease, 50
2~4h is digested at~60 DEG C, obtains enzymolysis liquid；The protease include alkali protease, neutral proteinase, acid protease,
Flavor protease or papain；
2) enzymolysis liquid for obtaining the step 1) carries out destroy the enzyme treatment, and obtained destroy the enzyme treatment liquid is centrifugated, will
Obtained supernatant is filtered, and obtains filtrate, and the speed of the centrifuge separation is 4000~5000rpm；
3) filtrate for obtaining the step 2) carries out first time ultrafiltration using the ultrafiltration membrane that molecular cut off is 10kDa,
Obtained first time ultrafiltrate is subjected to second of ultrafiltration with molecular cut off for the ultrafiltration membrane of 5kDa, obtains second of ultrafiltration
Liquid；
4) second of ultrafiltrate for obtaining the step 3) carries out third time with molecular cut off for the ultrafiltration membrane of 3kDa
Ultrafiltration, obtain peptide fragment molecular weight [3~5) the queen bee nit glucosidase suppressor activity peptide of KDa.
Preferably, after second of ultrafiltrate carries out third time ultrafiltration, by obtained third time ultrafiltrate to retain point
The ultrafiltration membrane that son amount is 1kDa carries out the 4th ultrafiltration, obtain peptide fragment molecular weight be [1~3) the queen bee nit glucoside of KDa
Enzyme inhibitory peptide.
Preferably, after the third time ultrafiltrate carries out the 4th ultrafiltration, by the obtain the 4th ultrafiltrate to retain point
The ultrafiltration membrane that son amount is 0.3~0.5kDa carries out the 5th ultrafiltration, obtains the queen bee nit glucose of peptide fragment molecular weight < 1kDa
Glycosides enzyme inhibitory peptide.
Preferably, the mass ratio of the enzyme activity of the step 1) protease and queen bee larva is (1500~2500) U:1g；
The time of the protease hydrolyzed is 2.5~3.5h.
Preferably, the quality of the step 1) queen bee larva and the volume ratio of water are 1g:(2~6) mL.
Preferably, the first time ultrafiltration, second of ultrafiltration, third time ultrafiltration, the 4th ultrafiltration and the 5th ultrafiltration
Conditional sampling includes: filtrate flow velocity for 0.5~10L/H, and membrane component pressure is 0~0.8MPa, and pure water is added before terminating in ultrafiltration
It is washed, until the permeate of outflow is colourless.
The present invention also provides a kind of queen bee nit glucosidase suppressor activity peptide, the queen bee nit glucuroide
The molecular weight of inhibitory activity peptide be [3~5) KDa.
The present invention also provides the queen bee nit glucosidase suppressor activity peptides described in above-mentioned technical proposal to drop in preparation
Application in hypoglycemic medicament.
The present invention also provides the queen bee nit glucosidase suppressor activity peptides described in above-mentioned technical proposal to eat in preparation
Application in product.
The present invention provides a kind of preparation method of queen bee nit glucosidase suppressor activity peptide, by queen bee larva through enzyme
After enzymatic hydrolysis, the mixtures such as different molecular weight peptide fragment, lipid like material, salt, insoluble sludge are obtained, after centrifuge separation,
The substances such as soluble peptide fragment, salt, insoluble lipid are left, insoluble sludge is cast out, after filtering, remove insoluble rouge
The impurity of matter class and partial size greater than 5 μm except desalting, leaves soluble peptide fragment, obtains queen bee nit glucoside after ultrafiltration
Enzyme inhibitory peptide.
The embodiment of the present invention is as the result is shown: preparation method provided by the invention can prepare molecular weight be [3~5) kDa
Active peptide obtain the peptide fragment of different molecular weight, middle-molecular-weihydroxyethyl is further using after the ultrafiltration of different molecular weight ultrafiltration membrane
[3~5) the inhibition glucoside enzyme performance highest that has of the active peptide of kDa.
Specific embodiment
The present invention provides a kind of preparation methods of queen bee nit glucosidase suppressor activity peptide, comprising the following steps:
1) queen bee larva is mixed into homogenate with water, after the pH value for adjusting homogenate is 6~7, is mixed with protease, 50
2~4h is digested at~60 DEG C, obtains enzymolysis liquid；The protease include alkali protease, neutral proteinase, acid protease,
Flavor protease or papain；
2) enzymolysis liquid for obtaining the step 1) carries out destroy the enzyme treatment, and obtained destroy the enzyme treatment liquid is centrifugated, will
Obtained supernatant is filtered, and obtains filtrate, and the speed of the centrifuge separation is 4000~5000rpm；
3) filtrate for obtaining the step 2) carries out first time ultrafiltration using the ultrafiltration membrane that molecular cut off is 10kDa,
Obtained first time ultrafiltrate is subjected to second of ultrafiltration with molecular cut off for the ultrafiltration membrane of 5kDa, obtains second of ultrafiltration
Liquid；
4) second of ultrafiltrate for obtaining the step 3) carries out third time with molecular cut off for the ultrafiltration membrane of 3kDa
Ultrafiltration, obtain peptide fragment molecular weight [3~5) the queen bee nit glucosidase suppressor activity peptide of KDa.
Queen bee larva is mixed homogenate with water by the present invention, after the pH value for adjusting homogenate is 6~7, is mixed with protease,
2~4h is digested at 50~60 DEG C, obtains enzymolysis liquid；The protease includes alkali protease, neutral proteinase, acidic protein
Enzyme, flavor protease or papain.
In the present invention, the queen bee nit is preferably honeybee fertilized eggs on the royal cell not covered, is fed with bee through worker bee
Royal jelly is after hatching to the larva body before nymphosis.Grinds again after preferably queen bee nit is lyophilized by the present invention, obtain queen bee nit
Powder, the present invention is not particularly limited the condition of the queen bee nit grinds of the freeze-drying, using conventional technical means.
In the present invention, the partial size of the queen bee larva is preferably 10~100 μm, more preferably 15~25 μm, optimal
It is selected as 20 μm.
In the present invention, the quality of the queen bee larva and the volume ratio of water are preferably 1g:(2~6) mL, more preferably
1g:(3~5) mL, most preferably 1g:4mL.
The present invention is not particularly limited the condition of the homogenate, using the conventional use of homogenate bee of those skilled in the art
The condition of king larva.
The pH value that the present invention adjusts homogenate is 6~7, preferably 6.2~6.8, more preferably 6.5.The present invention preferably uses
Dilute hydrochloric acid and sodium hydroxide solution adjust the pH value of homogenate.
In the present invention, the protease includes alkali protease, neutral proteinase, acid protease, flavor protease
Or papain.The source of protease enzyme of the present invention is not particularly limited, using conventional commercial product.
In the present invention, the mass ratio of the enzyme activity of the protease and queen bee larva is preferably (1500~2500) U:
1g, more preferably 1700~2300U:1g, most preferably 1900~2100U:1g.
It is mixed after the pH value of present invention adjusting homogenate with enzyme, 2~4h is digested at 50~60 DEG C, obtains enzymolysis liquid, it is excellent
It is selected in 2.5~3.5h of enzymatic hydrolysis at 52~58 DEG C, more preferably digests 3h at 53~55 DEG C.In the present invention, the queen bee nit
Powder obtains the mixtures such as different molecular weight peptide, lipoid material, salt, insoluble sludge after digesting.
Obtained enzymolysis liquid is carried out destroy the enzyme treatment by the present invention, obtained destroy the enzyme treatment liquid is centrifugated, by what is obtained
Supernatant is filtered, and obtains filtrate, and the speed of the centrifuge separation is 4000~5000rpm.
In the present invention, the condition of the destroy the enzyme treatment preferably includes: the time of the destroy the enzyme treatment is preferably 5~
15min, more preferably 8~12min, most preferably 10min；The temperature of the destroy the enzyme treatment is preferably 90~110 DEG C, more preferably
It is 95~105 DEG C, most preferably 100 DEG C.
In the present invention, the speed of the centrifuge separation be 4000~5000rpm, preferably 4200~4800rpm, it is more excellent
It is selected as 4500rpm；The time of the centrifuge separation is preferably 5~10min, more preferably 6~9min, most preferably 7~8min.
In the present invention, the centrifuge separation leaves the substances such as soluble peptides, salt, insoluble fats, casts out insoluble sludge.
In the present invention, the filtering is it is preferable to use filter cloth progress, and the aperture of the filter cloth is preferably 1~5 μm, more preferably
It is 2~4 μm, most preferably 3 μm.In the present invention, the filtering removes the impurity of insoluble fats and partial size greater than 5 μm.
In the present invention, the condition of the first time ultrafiltration preferably includes: filtrate flow velocity is preferably 0.5~10L/H, more excellent
It is selected as 2~8L/H；Membrane component pressure is preferably 0~0.8MPa, and ultrafiltration is added pure water before terminating and is washed, until outflow is saturating
It is colourless for crossing liquid.In the present invention, the peptide that the ultrafiltration makes ultrafiltrate middle-molecular-weihydroxyethyl be less than molecular cut off filters out as far as possible.
In the present invention, the peptide that the ultrafiltration makes ultrafiltrate middle-molecular-weihydroxyethyl be less than molecular cut off filters out as far as possible.
After first time ultrafiltrate is carried out second of ultrafiltration by the present invention, by second obtained of ultrafiltrate with molecular cut off
Carry out third time ultrafiltration for the ultrafiltration membrane of 3kDa, obtain peptide fragment molecular weight be [3~5) suppression of the queen bee nit glucuroide of KDa
Active peptide processed.
In the present invention, the condition of second of ultrafiltration preferably includes: filtrate flow velocity is preferably 0.5~10L/H, more excellent
It is selected as 2~8L/H；Membrane component pressure is preferably 0~0.8MPa, and ultrafiltration is added pure water before terminating and is washed, until outflow is saturating
It is colourless for crossing liquid.In the present invention, the peptide that the ultrafiltration makes ultrafiltrate middle-molecular-weihydroxyethyl be less than molecular cut off filters out as far as possible.
In the present invention, the peptide that the ultrafiltration makes ultrafiltrate middle-molecular-weihydroxyethyl be less than molecular cut off filters out as far as possible.
After second ultrafiltrate is preferably carried out third time ultrafiltration by the present invention, by obtained third time ultrafiltrate to retain point
The ultrafiltration membrane that son amount is 1kDa carries out the 4th ultrafiltration, obtain peptide fragment molecular weight be [1~3) the queen bee nit glucoside of KDa
Enzyme inhibitory peptide.
In the present invention, the condition of the 4th ultrafiltration preferably includes: filtrate flow velocity is preferably 0.5~10L/H, more excellent
It is selected as 2~8L/H；Membrane component pressure is preferably 0~0.8MPa, and ultrafiltration is added pure water before terminating and is washed, until outflow is saturating
It is colourless for crossing liquid.In the present invention, the peptide that the ultrafiltration makes ultrafiltrate middle-molecular-weihydroxyethyl be less than molecular cut off filters out as far as possible.
In the present invention, the peptide that the ultrafiltration makes ultrafiltrate middle-molecular-weihydroxyethyl be less than molecular cut off filters out as far as possible.
After third time ultrafiltrate is preferably carried out the 4th ultrafiltration by the present invention, by the obtain the 4th ultrafiltrate to retain point
The ultrafiltration membrane that son amount is 0.3~0.5kDa carries out the 5th ultrafiltration, obtains the queen bee nit grape that peptide fragment molecular weight is < 1kDa
Glucoside inhibiting activity peptide.
In the present invention, the condition of the 5th ultrafiltration preferably includes: filtrate flow velocity is preferably 0.5~10L/H, more excellent
It is selected as 2~8L/H；Membrane component pressure is preferably 0~0.8MPa, and ultrafiltration is added pure water before terminating and is washed, until outflow is saturating
It is colourless for crossing liquid.In the present invention, the peptide that the ultrafiltration makes ultrafiltrate middle-molecular-weihydroxyethyl be less than molecular cut off filters out as far as possible.
In the present invention, the peptide that the ultrafiltration makes ultrafiltrate middle-molecular-weihydroxyethyl be less than molecular cut off filters out as far as possible.
The present invention is freeze-dried after obtained ultrafiltrate is concentrated under reduced pressure respectively, obtains queen bee nit Glucosidase inhibitor
Active peptide.
The present invention is not particularly limited the condition of the reduced pressure and freeze-drying, normal using those skilled in the art
Advise the condition of the reduced pressure and freeze-drying that use.
The present invention also provides the queen bee nit glucosidase suppressor activity peptide described in above-mentioned technical proposal, the queen bees
The molecular weight of larva glucosidase suppressor activity peptide be [3~5) KDa.
The present invention also provides the queen bee nit Glucosidase inhibitors that the preparation method described in above scheme is prepared
Active peptide is preparing the application in hypoglycemic drug.In the present invention, the queen bee nit glucosidase suppressor activity peptide energy
Enough inhibit the activity of glucuroide.
The present invention also provides the queen bee nit Glucosidase inhibitors that the preparation method described in above scheme is prepared
Active peptide is preparing the application in food.In the present invention, the queen bee nit glucosidase suppressor activity peptide is able to suppress
The activity of glucuroide.
Below with reference to embodiment to a kind of preparation side of queen bee nit glucosidase suppressor activity peptide provided by the invention
Method and queen bee nit glucosidase suppressor activity peptide and application are described in detail, but they cannot be interpreted as to this
The restriction of invention protection scope.
Embodiment 1
Freeze-drying queen bee nit is milled, 80 meshes is crossed, is homogenized after the water of freeze-drying 4 times of volumes of queen bee nit weight is added, used
Dilute hydrochloric acid and low-concentration sodium hydroxide solution adjust pH to 6.5, and neutral proteinase is added, digests 3h, albumen under the conditions of 50 DEG C
The enzyme activity of enzyme and the mass ratio of queen bee larva are 2000U:1g, obtain enzymolysis liquid after enzymatic hydrolysis.
By enzymolysis liquid temperature be 100 DEG C at destroy the enzyme treatment 10min, by obtained destroy the enzyme treatment liquid 4000rpm item
Be centrifuged 6min under part, discard upper layer grease, supernatant is taken to cross 5 μm of filter clothes, by filtrate molecular cut off be 10kDa ultrafiltration membrane into
Row first time ultrafiltration, then second of ultrafiltration is carried out with the ultrafiltration membrane that molecular cut off is 5kDa, then with molecular cut off be 3kDa
Ultrafiltration membrane carry out third time ultrafiltration, then ultrafiltration membrane with molecular cut off for 1kDa carries out the 4th ultrafiltration, then with retention point
Son amount be 0.3~0.5kDa ultrafiltration membrane carries out the 5th ultrafiltration, respectively obtain peptide fragment molecular weight > 5kDa, [3~5) kDa, [1
~3) ultrafiltrate of kDa, < 1kDa, filtrate flow velocity is 5L/H in ultra-filtration process, and the membrane component pressure of the ultrafiltration is 0.4MPa,
Pure water is added before ultrafiltration terminates to be washed, until the permeate of outflow is colourless.
Be freeze-dried after obtained trapped fluid is concentrated under reduced pressure, obtain queen bee nit different molecular weight > 5kDa, [3~
5) kDa, [1~3) kDa, < 1kDa peptide fragment.
Measure obtain queen bee nit different molecular weight > 5kDa, [3~5) kDa, [1~3k) Da, < 1kDa peptide fragment with
And the glucosidase inhibitory active of enzymolysis liquid, > 5kDa, [3~5) kDa, [1~3) kDa, < 1kDa peptide fragment and enzymolysis liquid
503nhibiting concentration to glucuroide be respectively 45.51mg/ml, 34.64mg/ml, 37.39mg/ml, 36.29mg/ml,
49.47mg/ml.Illustrate after multiple ultrafiltration, the activity of small-molecular peptides is obviously improved than enzymolysis liquid, wherein molecular weight [3~
5) the peptide activity highest of kDa.
Embodiment 2
Freeze-drying queen bee nit is milled, 80 meshes is crossed, is homogenized after the water of freeze-drying 5 times of volumes of queen bee nit weight is added, used
Dilute hydrochloric acid and low-concentration sodium hydroxide solution adjust pH to 6.8, and neutral proteinase is added, digests 2.5h, egg under the conditions of 52 DEG C
The enzyme activity of white enzyme and the mass ratio of queen bee larva are 1900U:1g, obtain enzymolysis liquid after enzymatic hydrolysis.
By enzymolysis liquid temperature be 90 DEG C at destroy the enzyme treatment 15min, by obtained destroy the enzyme treatment liquid 4500rpm condition
Lower centrifugation 7min, discards upper layer grease, and supernatant is taken to cross 4 μm of filter clothes, and the ultrafiltration membrane that filtrate molecular cut off is 10kDa is carried out
First time ultrafiltration, then second of ultrafiltration is carried out with the ultrafiltration membrane that molecular cut off is 5kDa, then with molecular cut off be 3kDa's
Ultrafiltration membrane carries out third time ultrafiltration, then the ultrafiltration membrane with molecular cut off for 1kDa carries out the 4th ultrafiltration, then to retain molecule
Amount be 0.3~0.5kDa ultrafiltration membrane carry out the 5th ultrafiltration, respectively obtain peptide fragment molecular weight > 5kDa, [3~5) kDa, [1~
3) ultrafiltrate of kDa, < 1kDa, filtrate flow velocity is 3L/H in ultra-filtration process, and the membrane component pressure of the ultrafiltration is 0.2MPa,
Ultrafiltration is added pure water before terminating and is washed, until the permeate of outflow is colourless.
Be freeze-dried after obtained trapped fluid is concentrated under reduced pressure, obtain queen bee nit different molecular weight > 5kDa, [3~
5) kDa, [1~3) kDa, < 1kDa peptide fragment.
Measure obtain queen bee nit different molecular weight > 5kDa, [3~5) kDa, [1~3k) Da, < 1kDa peptide fragment with
And the glucosidase inhibitory active of enzymolysis liquid, > 5kDa, [3~5) kDa, [1~3) kDa, < 1kDa peptide fragment and enzymolysis liquid
503nhibiting concentration to glucuroide be respectively 46.82mg/ml, 33.25mg/ml, 38.58mg/ml, 38.10mg/ml,
52.36mg/ml.Illustrate after multiple ultrafiltration, the activity of small-molecular peptides is obviously improved than enzymolysis liquid, wherein molecular weight [3~
5) the peptide activity highest of kDa.
Embodiment 3
Freeze-drying queen bee nit is milled, 80 meshes is crossed, is homogenized after the water of freeze-drying 3 times of volumes of queen bee nit weight is added, used
Dilute hydrochloric acid and low-concentration sodium hydroxide solution adjust pH to 6.2, and neutral proteinase is added, digests 3.5h, egg under the conditions of 55 DEG C
The enzyme activity of white enzyme and the mass ratio of queen bee larva are 2100U:1g, obtain enzymolysis liquid after enzymatic hydrolysis.
By enzymolysis liquid temperature be 110 DEG C at destroy the enzyme treatment 5min, by obtained destroy the enzyme treatment liquid 4800rpm condition
Lower centrifugation 6min, discards upper layer grease, and supernatant is taken to cross 5 μm of filter clothes, and the ultrafiltration membrane that filtrate molecular cut off is 10kDa is carried out
First time ultrafiltration, then second of ultrafiltration is carried out with the ultrafiltration membrane that molecular cut off is 5kDa, then with molecular cut off be 3kDa's
Ultrafiltration membrane carries out third time ultrafiltration, then the ultrafiltration membrane with molecular cut off for 1kDa carries out the 4th ultrafiltration, then to retain molecule
Amount be 0.3~0.5kDa ultrafiltration membrane carry out the 5th ultrafiltration, respectively obtain peptide fragment molecular weight > 5kDa, [3~5) kDa, [1~
3) ultrafiltrate of kDa, < 1kDa, filtrate flow velocity is 8L/H in ultra-filtration process, and the membrane component pressure of the ultrafiltration is 0.7MPa,
Ultrafiltration is added pure water before terminating and is washed, until the permeate of outflow is colourless.
Be freeze-dried after obtained trapped fluid is concentrated under reduced pressure, obtain queen bee nit different molecular weight > 5kDa, [3~
5) kDa, [1~3) kDa, < 1kDa peptide fragment.
Measure obtain queen bee nit different molecular weight > 5kDa, [3~5) kDa, [1~3k) Da, < 1kDa peptide fragment with
And the glucosidase inhibitory active of enzymolysis liquid, > 5kDa, [3~5) kDa, [1~3) kDa, < 1kDa peptide fragment and enzymolysis liquid
503nhibiting concentration to glucuroide be respectively 47.26mg/ml, 35.43mg/ml, 39.14mg/ml, 38.49mg/ml,
50.22mg/ml.Illustrate after multiple ultrafiltration, the activity of small-molecular peptides is obviously improved than enzymolysis liquid, wherein molecular weight [3~
5) the peptide activity highest of kDa.
As seen from the above embodiment, the preparation side of a kind of queen bee nit glucosidase suppressor activity peptide provided by the invention
Method, can prepare [3~5) active peptide of kDa further using after the ultrafiltration of different molecular weight ultrafiltration membrane obtains different molecular
The peptide fragment of amount, middle-molecular-weihydroxyethyl be [3~5) the inhibition glucoside enzyme performance highest that has of the active peptide of kDa.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (4)
1. a kind of preparation method of queen bee nit glucosidase suppressor activity peptide, comprising the following steps:
1) queen bee larva is crossed after 80 meshes and mixes homogenate with water, the volume ratio of the quality of queen bee larva and water be 1g:3~
5mL is mixed, the mass ratio of enzyme activity and queen bee larva is after the pH value for adjusting homogenate is 6.2~6.8 with neutral proteinase
1900~2100U:1g digests 2.5~3.5h at 50~55 DEG C, obtains enzymolysis liquid；
2) enzymolysis liquid for obtaining the step 1) carries out destroy the enzyme treatment, and obtained destroy the enzyme treatment liquid is centrifugated, will be obtained
Supernatant cross 4 μm of filter clothes, obtain filtrate, the speed of the centrifuge separation is 4000~4800rpm；
3) filtrate for obtaining the step 2) carries out first time ultrafiltration using the ultrafiltration membrane that molecular cut off is 10kDa, will
The first time ultrafiltrate arrived carries out second of ultrafiltration with molecular cut off for the ultrafiltration membrane of 5kDa, obtains second of ultrafiltrate；
4) second of ultrafiltrate for obtaining the step 3) carries out third time ultrafiltration with molecular cut off for the ultrafiltration membrane of 3kDa,
Obtain peptide fragment molecular weight [3~5) the queen bee nit glucosidase suppressor activity peptide of kDa；
Include: to the conditional sampling of the first time ultrafiltration, second of ultrafiltration and third time ultrafiltration filtrate flow velocity be 3~8L/h,
Membrane component pressure is 0.2~0.7MPa, and ultrafiltration is added pure water before terminating and is washed, until the permeate of outflow is colourless.
2. a kind of queen bee nit glucosidase suppressor activity peptide of preparation method preparation as described in claim 1.
3. a kind of queen bee nit glucosidase suppressor activity peptide as claimed in claim 2 reduces in hypoglycemic medicament in preparation
Using.
4. a kind of queen bee nit glucosidase suppressor activity peptide as claimed in claim 2 is preparing the application in food.
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| CN106937731A (en) * | 2016-01-05 | 2017-07-11 | 上海辉文生物技术股份有限公司 | Queen bee nit powder extracts and its preparation method and application |
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| CN106937731A (en) * | 2016-01-05 | 2017-07-11 | 上海辉文生物技术股份有限公司 | Queen bee nit powder extracts and its preparation method and application |
Non-Patent Citations (3)
| Title |
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| 响应面法优化蚕蛹蛋白源α-葡萄糖苷酶抑制肽酶解条件，张玉，《中国食品学报》，第16卷第4期，第137-144页;张玉;《中国食品学报》;20160430;第16卷(第4期);摘要，引文，方法，结论 |
| 蜂王幼虫活性肽保健功能的研究;陈明胜;《2008年全国蜂产品市场信息交流大会论文集》;20081231;全文 |
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