CN107099571A - A kind of bionic enzymatic prepares the production method of silkworm chrysalis small peptide - Google Patents
A kind of bionic enzymatic prepares the production method of silkworm chrysalis small peptide Download PDFInfo
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- CN107099571A CN107099571A CN201710295493.XA CN201710295493A CN107099571A CN 107099571 A CN107099571 A CN 107099571A CN 201710295493 A CN201710295493 A CN 201710295493A CN 107099571 A CN107099571 A CN 107099571A
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- silkworm chrysalis
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Classifications
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- C—CHEMISTRY; METALLURGY
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
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- A—HUMAN NECESSITIES
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- A23L33/18—Peptides; Protein hydrolysates
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Abstract
The invention discloses the industrial process that a kind of bionic enzymatic from pupa albumen produces silkworm chrysalis small peptide.It is characterized in that using pupa albumen as starting material, by bionic enzymatic, high speed centrifugation, ultrafiltration, electrodialysis, concentrate, be dried to obtain silkworm chrysalis small peptide.The silkworm chrysalis small peptide total protein content more than 90%, the soluble small-molecular peptides content of trichloroacetic acid more than 85%, with meet absorption of human body selection mechanism, have no toxic side effect, low cost and other advantages, can be applied in food, health food and medicine.
Description
Technical field
The present invention relates to a kind of preparation method of silkworm chrysalis small peptide, and in particular to one kind bionic enzymatic from pupa albumen is produced
The industrial process of silkworm chrysalis small peptide.
Background technology
Silkworm chrysalis is exactly nourishing and fit keeping function from ancient times, gentle spleen kidney, the medicine and food of long muscle.Silkworm chrysalis has extremely abundant battalion
Value and medical value are supported, it contains abundant protein and essential amino acid, electrolytes and minerals, be mankind's reason
The animal food thought.Pupa albumen is the silkworm chrysalis pulverulent solids that pulverization process is obtained after dehydration, grease removal.The albumen of silkworm chrysalis
Matter content is more than 50%, the essential amino acid A wide selection of colours and designs significantly larger than in normal food, and protein, silkworm chrysalis protein by
18 kinds of amino acid compositions, wherein 8 kinds of amino acid contents needed by human are very high, it is 2 times of pork, 4 times of egg, the 10 of milk
Times, 8 kinds of amino acid nutrients needed by human are balanced, and ratio is appropriate, meets FAO/WHO (FAO (Food and Agriculture Organization of the United Nation) and world health
Tissue) requirement；Silkworm chrysalis also containing potassium, receive, calcium, magnesium, iron, copper, manganese, zinc, phosphorus, selenium and other trace elements, vitamin A, E, B1,
B2, carrotene etc.；The content of unrighted acid in silkworm chrysalis is enriched very much, accounts for the 72.5% of total fat.But pupa albumen
Molecular weight is larger, and human body can not directly absorb, and need to can just play corresponding biological living for the peptide of small molecule by twice decomposition in vivo
Property.
Bionic enzymatic be simulate oral protein pipe intestinal digesting process (stomach be Digestive site, absorb seldom, enteron aisle is
Digestion, absorption site), the creative method that protide is digested into small-molecule peptide, i.e., under suitable condition with stomach egg
White enzyme, trypsase carry out continuous enzymolysis, then obtain small with relative molecular mass through high speed centrifugation, ultrafiltration, electrodialysis or nanofiltration
In the small-molecule peptide based on 3KD.Compared with conventional art, the active materials of absorbed into serum are identical with direct oral protein,
Bionic enzymatic extracts active ingredients are more abundant, and bioavilability is high, not only increases curative effect, and eliminate and take M-band etc.
Caused immunogene reaction, reduces adverse reaction rate, safe；Compared with other enzyme solutions, bionic enzymatic is more
Meet the selection mechanism of human body, it is safer.
The inventive method that silkworm chrysalis small peptide is inventor, the work of amplifiedization are produced from pupa albumen in bionic enzymatic method
Industry production checking, it is stable, feasible, have broad application prospects.
The content of the invention
The first object of the present invention is to provide the industry life that a kind of bionic enzymatic from pupa albumen produces silkworm chrysalis small peptide
Production method；Second purpose is to provide the silkworm chrysalis small peptide obtained by described industrialized preparing process；3rd purpose is to carry
Take application of the described silkworm chrysalis small peptide in food, health food and medicine field.
The object of the present invention is achieved like this：
A kind of bionic enzymatic from pupa albumen produces the industrial process of silkworm chrysalis small peptide, using following steps：
(1) add purified water 20-40 liters into bionic enzymatic tank, start stirring slurry, 30-60 revs/min of rotating speed is set, silkworm is taken
Pupa albumen powder 2kg, is slowly added to, and addition continues to stir swollen 30 minutes after finishing, and interlayer is logical to be steam heated to 80-85 DEG C of sterilizing
15-30 minutes, interlayer led to cooling and is water-cooled to about 50-55 DEG C of temperature in tank, starts intermediate water and is circulated with heater, heater adds
Hot to 50-55 DEG C insulation；
(2) start pH detection terminals, watery hydrochloric acid regulation pH value 1.8-2.2 is added with peristaltic pump into bionic enzymatic tank, plus
Enter the 1-3% pepsins of pupa albumen amount, insulation enzymolysis 1-3 hours monitors pH value at any time, and supplement watery hydrochloric acid keeps pH value to exist
1.8-2.2 in the range of；
(3) 40% sodium hydroxide regulation pH value 7.8-8.2 is added with peristaltic pump into bionic enzymatic tank, adds 1-3% pancreases
Protease, insulation enzymolysis 3-6 hours monitors pH value at any time, and 40% sodium hydroxide of supplement keeps pH value in the range of 7.8-8.2；
(4) watery hydrochloric acid regulation pH value 6.8-7.2 is added with peristaltic pump into bionic enzymatic tank, bleeds off intermediate water, interlayer leads to
Steam, which boils, is heated to 80-85 DEG C of enzyme 15-30 minutes of going out, tapping；
(5) centrifuged into tubular-bowl centrifuge, 8000~12000 revs/min of rotating speed, supernatant passes through 1812 type ultrafiltration
Device ultrafiltration, milipore filter relative molecular mass is 3KD, and permeate is through electrodialysis desalination, 30~90 volts of voltage, except saline solution is through decompression
The clear cream that 60 DEG C of relative densities are 1.10~1.15 is concentrated into, spray drying, EAT is 180-200 DEG C, leaving air temp is
90-100 DEG C, obtain silkworm chrysalis small peptide, compound membrane bag packaging.
Protein content is more than 90% in pupa albumen described in step (1), and moisture is less than 10%；
Pepsin enzyme activity described in step (2) is 3000U/g；
Trypsase enzyme activity described in step (3) is 100000U/g；
Total protein content is more than 90% in silkworm chrysalis small peptide described in step (5), the soluble small molecule of trichloroacetic acid
Peptide content is more than 85%, and free aminoacid content is less than 5%, and moisture is less than 7%.
Mass spectroscopy is carried out to silkworm chrysalis small peptide in the present invention, small peptide peptide chain structure confirmed, small peptide mass spectrum (ESI-
MS) assay method and result are as follows:
ESI-MS parameters atomization gas is nitrogen, and collision gas are argon gas；80 DEG C of source temperature, taper hole voltage 50V；TOF accelerates electricity
Press 9.1kV；MCP detector voltages are 2150V；Capillary voltage 800V.
Analyzed first, mass scan range is 100~1000D.Shown according to the measurement result of this product, most of peptide
Duan Erchong protonates (i.e. two unit positive charges of band), is analyzed by the strong parent ion of selection signal into MS/MS, obtains 8 kinds
The MS/MS collection of illustrative plates of peptide, is analyzed obtained MS/MS patch informations, and their the amino acid sequence composition result of acquisition is shown in
Table 1.
The silkworm chrysalis small peptide relative molecular mass of table 1 and amino acid sequence composition table
Note：Sequence letter represents writing a Chinese character in simplified form for different aminoacids, alanine A, arginine R, aspartic acid D, half Guang ammonia in table
Sour C, glutamine Q, glutamic acid E, histidine H, isoleucine I, glycine G, asparagine N, leucine L, lysine K, first
Methyllanthionine M, phenylalanine F, proline P, serine S, threonine T, tryptophan W, tyrosine Y, valine V.
Prepare silkworm chrysalis small peptide using bionic enzymatic method, not only increase curative effect, and eliminate take the former powder of animal and its
His general extraction methods introduce immunogene reaction caused by M-band, reduce adverse reaction rate, safe.
Because the position of different enzyme hydrolysis is different, obtained enzymolysis product also differs widely, and such as pepsin is main
The position of enzymolysis be phenylalanine, tyrosine and leucine composition peptide bond, the position that trypsase is mainly digested be arginine,
The peptide bond of the basic amino acids such as lysine composition.
This research is with a batch of silkworm chrysalis medicinal material, using total peptide amount as index, has investigated silkworm chrysalis using different protease enzymes
The activity of the extract solution of technique is solved, that is, has investigated silkworm chrysalis pepsin enzymolysis, trypsin digestion, papain enzymolysis and has imitated
The content of enzymolysis liquid obtained by raw enzymatic isolation method.
Silkworm chrysalis medicinal material is taken, fine powder is ground into, four parts are weighed, every part of 20g, plus 10 times of amount water boil 15 minutes, wait to be cooled to 40
DEG C, it is a that pH value 1.5~2.0 is adjusted with dilute hydrochloric acid solution, the pepsin of 2% amount is added, 40 DEG C of insulations are digested 2 hours；One
Part adjusts pH value 8.0~8.5 with 20% sodium hydroxide solution, adds the trypsase of 2% amount, and 40 DEG C of insulations are digested 4 hours；One
Part adjusts the papain that pH value 6.0~7.0 adds 2% amount with 20% sodium hydroxide solution, and 40 DEG C of insulations are digested 4 hours；
Last a pepsin that pH value 1.5~2.0 is adjusted with dilute hydrochloric acid solution, 1% amount is added, 40 DEG C are incubated enzymolysis 2 hours,
20% sodium hydroxide solution adjusts pH value 8.0~8.5, adds the trypsase of 1% amount, and 40 DEG C of insulations are digested 4 hours；Above enzyme
Solution liquid heats to boil and makes enzyme-deactivating, centrifuges (5000r/min), supernatant be concentrated under reduced pressure (70~80 DEG C, -0.07~-
0.08MPa) and total amount is adjusted to 100mL, produce the different zyme extracts of silkworm chrysalis.Total peptide amount of each extract solution is determined respectively, is determined
It the results are shown in Table 2.
The component content of the silkworm chrysalis of table 2 difference protease hydrolyzed extracting method each several part
Test result indicates that, digested with single pepsin, trypsase, the albumen in silkworm chrysalis can not had
The utilization of effect, the activity and total peptide content of enzymolysis product be not high, and display medicinal material is underused, and causes certain medicinal material wave
Take；Double enzyme continuous enzymolysis and papain enzymolysis its lytic activity and total peptide content are higher, and double enzyme continuous enzymolysis are higher than wood
Melon protease, but because papain enzymolysis force enzymolysis in vitro, with the active material obtained by human body itself enzymolysis not to the utmost
It is identical, the rejection of human body must be caused.
A kind of silkworm chrysalis small peptide, is obtained by above-mentioned industrialized preparing process.The silkworm chrysalis small peptide obtained using this method,
Its main relative molecular mass is below 3KD, and with extremely strong water solubility, its total protein content is more than 90%, trichloroacetic acid
Soluble small-molecular peptides content is more than 85%, and free aminoacid content is less than 5%, and moisture is less than 7%.
Above-mentioned bionic enzymatic tank and enzymatic hydrolysis system refer to the equipment being related in CN205874433U, CN205590709U,
With the increase of volume of equipment, the consumption of each material can increase therewith in the inventive method.There is setting for similar function at other
Standby upper, this method is equally adapted to.
Raw material Silkworm pupa protein in the present invention, can be available from the finished product raw material of in the market or first from silkworm chrysalis
Reclaimed water solution obtains Silkworm pupa protein or the fluid product containing pupa albumen.In a word, no matter with which kind of raw material feed intake production (silkworm chrysalis,
Silkworm pupa protein, pupa albumen liquid etc.), as long as being related to the content of the invention protected in production technology, that is, it is considered as and employs
The technology contents of the present invention.
Application of the silkworm chrysalis small peptide obtained by above-mentioned industrial process in food, health food and medicine, including but
It is not limited to following applications：According to nutritive peculiarity, functional characteristic and the processing characteristics of the silkworm chrysalis small peptide produced, made with silkworm chrysalis small peptide
For basic material, many high-quality, nutrition, the food of health can be developed.Promote protein synthesis and antifatigue using it
Characteristic, can develop the movement nutrition food and health food of enhancing muscle and dispelling fatigue；Promote fat metabolism using it
Characteristic, can develop the weight losing function food for promoting fat metabolism；Promote the characteristic of cell growth using it, can be developed as
Promote microorganism growth, fermentation accelerant of metabolism etc..
The beneficial effects of the invention are as follows：Present invention bionic enzymatic from pupa albumen produces the industrial production side of silkworm chrysalis small peptide
Method, meets human body selection mechanism, and preparation technology is easy, feasible, and cost is low, production process environmental protection；The silkworm chrysalis small peptide of gained
Total protein content is more than 90%, and small-molecule peptide content is more than 85%, and relative molecular mass is nontoxic mainly in below 3KD
Side effect, reaches the pure natural requirement of pollution-free food, can obtain good economic benefit.
Embodiment
Embodiment is set forth below, the present invention is further illustrated, each embodiment is merely to illustrate the present invention, is not intended to limit this hair
It is bright：
The present embodiment of embodiment 1 comprises the following steps：
(1) add 20 liters of purified water into bionic enzymatic tank, start stirring slurry, 60 revs/min of rotating speed is set, pupa albumen is taken
(silkworm separated protein, protein content is more than 90%, and moisture is less than 10%) 2kg, is slowly added to, and addition continues to stir after finishing for powder
Mix swollen 30 minutes, interlayer is logical to be steam heated to 80-85 DEG C and sterilize 15 minutes, and interlayer leads to cooling and is water-cooled to temperature about 55 in tank
DEG C, start intermediate water and circulated with heater, heater is heated to 55 DEG C of insulations；
(2) start pH detection terminals, watery hydrochloric acid regulation pH value 2.0 is added with peristaltic pump into bionic enzymatic tank, silkworm is added
1% pepsin (enzyme activity is 3000U/g) of pupa protein content, insulation enzymolysis 1 hour, monitors pH value at any time, supplements watery hydrochloric acid
PH value is kept in the range of 1.8-2.2；
(3) 40% sodium hydroxide regulation pH value 8.0 is added with peristaltic pump into bionic enzymatic tank, adds 1% trypsase
(enzyme activity is 100000U/g), insulation enzymolysis 3 hours monitors pH value at any time, and 40% sodium hydroxide of supplement keeps pH value in 7.8-
8.2 in the range of；
(4) watery hydrochloric acid regulation pH value 6.8 is added with peristaltic pump into bionic enzymatic tank, bleeds off intermediate water, interlayer leads to steam
Boil and be heated to the 80-85 DEG C of enzyme 30 minutes of going out, tapping；
(5) centrifuged into tubular-bowl centrifuge, 8000~12000 revs/min of rotating speed, supernatant passes through 1812 type ultrafiltration
Device ultrafiltration, milipore filter relative molecular mass is 3KD, and permeate is through electrodialysis desalination, 30 volts of voltage, except saline solution is through being concentrated under reduced pressure
The clear cream for being 1.10~1.15 to 60 DEG C of relative densities, spray drying, EAT is 180-200 DEG C, and leaving air temp is 90-
100 DEG C, obtain silkworm chrysalis small peptide, compound membrane bag packaging.
The present embodiment of embodiment 2 comprises the following steps：
(1) add 40 liters of purified water into bionic enzymatic tank, start stirring slurry, 30 revs/min of rotating speed is set, pupa albumen is taken
Powder 2kg, is slowly added to, and addition continues to stir swollen 30 minutes after finishing, and interlayer is logical to be steam heated to 80-85 DEG C of 15 points of sterilizing
Clock, interlayer leads to cooling and is water-cooled to about 50 DEG C of temperature in tank, starts intermediate water and is circulated with heater, heater is heated to 50 DEG C of guarantors
Temperature；
(2) start pH detection terminals, watery hydrochloric acid regulation pH value 1.8 is added with peristaltic pump into bionic enzymatic tank, silkworm is added
3% pepsin of pupa protein content, insulation enzymolysis 3 hours monitors pH value at any time, and supplement watery hydrochloric acid keeps pH value in 1.8-2.2
In the range of；
(3) 40% sodium hydroxide regulation pH value 8.2 is added with peristaltic pump into bionic enzymatic tank, adds 3% trypsase,
Insulation enzymolysis 6 hours, monitors pH value at any time, and 40% sodium hydroxide of supplement keeps pH value in the range of 7.8-8.2；
(4) watery hydrochloric acid regulation pH value 7.2 is added with peristaltic pump into bionic enzymatic tank, bleeds off intermediate water, interlayer leads to steam
Boil and be heated to the 80-85 DEG C of enzyme 15 minutes of going out, tapping；
(5) centrifuged into tubular-bowl centrifuge, 8000~12000 revs/min of rotating speed, supernatant passes through 1812 type ultrafiltration
Device ultrafiltration, milipore filter relative molecular mass is 3KD, and permeate is through electrodialysis desalination, 90 volts of voltage, except saline solution is through being concentrated under reduced pressure
The clear cream for being 1.10~1.15 to 60 DEG C of relative densities, spray drying, EAT is 180-200 DEG C, and leaving air temp is 90-
100 DEG C, obtain silkworm chrysalis small peptide, compound membrane bag packaging.
The silkworm chrysalis small peptide of embodiment 3：
The silkworm chrysalis small peptide that industrialized preparing process described in Example 1 is obtained, after testing：Its total protein content is
91%, the soluble small-molecular peptides content of trichloroacetic acid is 92%, and free aminoacid content is 2.1%, and moisture is
4.3%, relative molecular weight distribution is：
| Relative molecular weight scope | Peak area percent |
| ＞ 3000D | 3.1% |
| 3000-2000D | 25.2% |
| 2000-1000D | 36.8% |
| 1000-300D | 27.5% |
| ＜ 300D | 7.4% |
Product prepared by the silkworm chrysalis small peptide of embodiment 4：
Silkworm chrysalis small peptide 400g obtained by the method for Example 1, by silkworm chrysalis small peptide:Corn oil:Beeswax:Lecithin is
10:36:3:1 mixed material, is ground 30 minutes through colloid mill, and 100 are pressed with gelatin, glycerine, purified water, iron oxide black：40：100：
1 ratio making soft capsule hide glue gelatin, 55 ± 5 DEG C of control glue box temperature, 40 ± 5 DEG C of heat insulation tank, 36 ± 5 DEG C of sprinkler body temperature, with
Pellet press suppresses soft capsule, per the tolerant heavy 0.50g of intragranular, 18 DEG C~26 DEG C cold air drying 4h (relative humidity is 30%~40%)
Sizing, washes ball, drying, selects ball, packaging, be prepared into soft capsule finished product.
Product prepared by the silkworm chrysalis small peptide of embodiment 5：
Silkworm chrysalis small peptide 100g obtained by the method for Example 1, adds corn oligopeptide powder 10g, wheat oligopeptide 10g, big
10g/ bags, solid beverage is made in beans Gly-His-Lys 10g, plus appropriate Sucralose, mixed material, packing.
Claims (5)
1. a kind of bionic enzymatic from pupa albumen produces the industrial process of silkworm chrysalis small peptide, it is characterised in that use following step
Suddenly：
（1）Add purified water 20-40 liters into bionic enzymatic tank, start stirring slurry, 30-60 revs/min of rotating speed is set, silkworm chrysalis egg is taken
White powder 2kg, is slowly added to, and addition continues to stir swollen 30 minutes after finishing, and interlayer is logical to be steam heated to 80-85 DEG C of sterilizing 15-
30 minutes, interlayer led to cooling and is water-cooled to about 50-55 DEG C of temperature in tank, starts intermediate water and is circulated with heater, heater heating
To 50-55 DEG C of insulation；
（2）Start pH detection terminals, watery hydrochloric acid regulation pH value 1.8-2.2 is added with peristaltic pump into bionic enzymatic tank, silkworm is added
The 1-3% pepsins of pupa protein content, insulation enzymolysis 1-3 hours monitors pH value at any time, and supplement watery hydrochloric acid keeps pH value in 1.8-
2.2 in the range of；
（3）40% sodium hydroxide regulation pH value 7.8-8.2 is added with peristaltic pump into bionic enzymatic tank, 1-3% trypsase is added,
Insulation enzymolysis 3-6 hours, monitors pH value at any time, and 40% sodium hydroxide of supplement keeps pH value in the range of 7.8-8.2；
（4）Watery hydrochloric acid regulation pH value 6.8-7.2 is added with peristaltic pump into bionic enzymatic tank, intermediate water is bled off, interlayer leads to steam
Boil and be heated to 80-85 DEG C of enzyme 15-30 minutes of going out, tapping；
（5）Centrifuged into tubular-bowl centrifuge, 8000-12000 revs/min of rotating speed, supernatant passes through 1812 type ultrafiltration apparatus
Ultrafiltration, milipore filter relative molecular mass is 3KD, and permeate is through electrodialysis desalination, voltage 30-90 volts, except saline solution is through being concentrated under reduced pressure
The clear cream for being 1.10-1.15 to 60 DEG C of relative densities, spray drying, EAT is 180-200 DEG C, and leaving air temp is 90-100
DEG C, obtain silkworm chrysalis small peptide, compound membrane bag packaging.
2. the bionic enzymatic according to claim 1 from pupa albumen produces the industrial process of silkworm chrysalis small peptide, it is special
Levy and be that described pepsin enzyme activity is 3000U/g；Trypsase enzyme activity is 100000U/g；Albumen in pupa albumen
Content is more than 90%, and moisture is less than 10%.
3. the bionic enzymatic according to claim 1 from pupa albumen produces the industrial process of silkworm chrysalis small peptide, it is special
Levy and be in resulting silkworm chrysalis small peptide that total protein content is more than 90%, the soluble small-molecular peptides content of trichloroacetic acid is 85%
More than, free aminoacid content is less than 5%, and moisture is less than 7%.
4. a kind of silkworm chrysalis small peptide, is obtained by the industrialized preparing process described in claim 1-3.
5. application of the silkworm chrysalis small peptide in food, health food and medicine field described in claim 4.
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| CN115669928A (en) * | 2022-11-02 | 2023-02-03 | 广东省农业科学院蚕业与农产品加工研究所 | Nutritional food and preparation method and application thereof |
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