CN107217082A - A kind of deer hemalbumin polypeptide and application - Google Patents
A kind of deer hemalbumin polypeptide and application Download PDFInfo
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- CN107217082A CN107217082A CN201710414019.4A CN201710414019A CN107217082A CN 107217082 A CN107217082 A CN 107217082A CN 201710414019 A CN201710414019 A CN 201710414019A CN 107217082 A CN107217082 A CN 107217082A
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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
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Abstract
The invention discloses a kind of deer hemalbumin polypeptide, its preparation method includes successively：Washed corpuscles, ultrasonic wave added haemolysis, microwave radiation technology alkali enzyme enzymolysis, microwave radiation technology papain enzymolysis and concentration step, the molecule have preferable homogeneity, and the accounting of oligopeptides is high, it is easy to absorb；The deer hemalbumin polypeptide has preferable antifatigue and antioxidation activity, with preferable effect；The deer hemalbumin polypeptide, which is used to prepare, has higher stability after health preserving wine, is not likely to produce precipitation or muddy, has preferable application prospect in field of health care products.
Description
Technical field
The present invention relates to health treatment technical field, more particularly to a kind of deer hemalbumin polypeptide and application.
Background technology
Deer blood refers to the fine and soft blood and thorax blood of deer animal such as sika deer and red deer, and research shows, deer blood have enhancing it is immune,
Enrich blood, anti-aging, it is antifatigue, improve sexual function, strengthening the essence, calm the nerves, promote to be metabolized and the function such as wound healing.For many years, I
State is only limitted to the utilization to former blood and processing for the utilization of deer blood resource, and wherein most is deposited in the form of deer-blood wine
.And in some developed countries, had begun to utilize animal blood enzymolysis exploitation peptides reagent and medicine.Therefore, deer is developed
Hemalbumin product has great importance to the exploitation of deer blood resource.
The complex chemical composition of deer blood, physical and chemical testing shows that fresh deer blood moisture is 80%~81%, and inorganic constituents is accounted for
2%~4%, ash content accounts for 3%~4%, organic principle 16%~17%.Wherein it is mainly in protein and a variety of enzymes, protein
Rich in 19 kinds of amino acid；In addition also containing a variety of lipids, free fatty, sterol, glycolipid, phosphatide, hormone, purine, vitamin and
Polysaccharide etc., and containing a variety of constants and healthy trace elements with household.
Deer blood protein peptide refers to the protein in deer blood is hydrolyzed to the polypeptide produced in the presence of protease.Polypeptide has
Extremely strong activity and diversity, fully adjustable human body physiological function, enhancing Human Physiology activity；Polypeptide can not only provide human body
Grown required nutriment, and with special biological function, absorption of human body is more easy to than protein.
Deer-blood wine on the market is largely scattered in high wine using fresh deer blood at present, in storage and transport process
In, with wine body segregation phenomenon occurs for deer blood, to all generating strong influence in outward appearance and quality, causes unnecessary damage
Lose.
The content of the invention
In order to overcome the deficiencies in the prior art, an object of the present invention is that a kind of deer hemalbumin of high oligopeptides ratio is more
Peptide.
The second object of the present invention is the application for providing the deer hemalbumin polypeptide.
An object of the present invention adopts the following technical scheme that realization：
A kind of deer hemalbumin polypeptide, its preparation method includes successively：
Washed corpuscles：Anti-coagulants is added into fresh deer blood, is disperseed, is obtained anti-freezing blood sample, anti-freezing blood sample is centrifuged, abandoned
Blood plasma is removed, haemocyte is collected；
Ultrasonic wave added haemolysis：By haemocyte brine, the distilled water of 1-2 times of volume is added, is adjusted with dilute NaOH
Section pH is 9-10, and using 35-37 DEG C of ultrasonic wave added haemolysis, supernatant is collected in centrifugation；
Microwave radiation technology alkali enzyme is digested：Distilled water diluting is added into supernatant into 8-10g haemocytes/L, basic protein is added
Enzyme, makes enzyme concentration be 6000-7000U/g haemocytes, 50-60 DEG C of microwave radiation technology enzymolysis, wherein, plus dilute NaOH solution makes supernatant
PH value keep 9-10, obtain enzymolysis liquid；
Microwave radiation technology papain enzymolysis：Adding watery hydrochloric acid makes enzymolysis liquid pH value be 6-7, adds papain, makes
Enzyme concentration is 5000-6000U/g haemocytes, and 50-60 DEG C of microwave radiation technology is digested, and enzymolysis liquid is heated into 90-95 DEG C of inactivation,
Centrifugation, isolates supernatant；
Concentration：With 22 μm of microporous barriers by supernatant liquid filtering, Brix is concentrated under reduced pressure into for 50-60 °, deer hemalbumin polypeptide is produced
Paste liquid.
Preferably, in anti-freezing step, the anti-coagulants is the mixed solution of citric acid, sodium citrate and glucose, its
In, the weight ratio of citric acid, sodium citrate and glucose is (0.8-1):(0.3-0.6):1.
Preferably, in washed corpuscles step, centrifugation rate is 3000-3500rpm, centrifugation time is 10-15min.
Preferably, in ultrasonic wave added haemolysis step, ultrasonic power is 300-400W.
Preferably, in microwave radiation technology alkali enzyme enzymolysis step, microwave frequency is 1500-3000MHz, power is 5kW.
Preferably, in microwave radiation technology alkali enzyme enzymolysis step, dilute NaOH concentration is 0.5mol/L.
Preferably, in microwave radiation technology papain enzymolysis step, dilute HCl concentration is 0.5mol/L.
Preferably, in concentration step, it is described to be filtered into micro porous filtration.
Preferably, the aperture of micro porous filtration is 22 μm.
The second object of the present invention adopts the following technical scheme that realization：
Application of the above-mentioned deer hemalbumin polypeptide on health products are prepared.
Compared with prior art, the beneficial effects of the present invention are：
(1) molecule for the deer hemalbumin polypeptide that the present invention is provided has preferable homogeneity, and the accounting of oligopeptides is high, it is easy to inhale
Receive；The deer hemalbumin polypeptide has preferable antifatigue and antioxidation activity, with preferable effect；
(2) the deer hemalbumin polypeptide that provides of the present invention has higher stability after being used to preparing health preserving wine, is difficult production
Raw precipitation is muddy, has preferable application prospect in field of health care products.
Embodiment
Below, with reference to embodiment, the present invention is described further, it is necessary to which explanation is, what is do not collided
Under the premise of, new embodiment can be formed between various embodiments described below or between each technical characteristic in any combination.
In detailed description below, such as non-specified otherwise, the reagent or material used can be by commercially available or conventional
The mode of research technique is obtained.
The present invention provides a kind of deer hemalbumin polypeptide, and its preparation method includes successively：
Washed corpuscles：Anti-coagulants is added into fresh deer blood, is disperseed, is obtained anti-freezing blood sample, anti-freezing blood sample is centrifuged, abandoned
Blood plasma is removed, haemocyte is collected；
Anti-coagulants is preferably the mixed solution that the anti-coagulants is citric acid, sodium citrate and glucose in this step, its
In, the weight ratio of citric acid, sodium citrate and glucose is (0.8-1):(0.3-0.6):1, i.e., the mixed type anti-coagulants can be more
Mend the single anti-coagulants defect such as cause system pH unstable；
Ultrasonic wave added haemolysis：By haemocyte brine, the distilled water of 1-2 times of volume is added, is adjusted with dilute NaOH
Section pH is 9-10, and using 35-37 DEG C of ultrasonic wave added haemolysis, supernatant is collected in centrifugation；
Using ultrasonic wave added, the dissolution rate of haemocyte is effectively improved；Under the pH value condition, haemocyte osmotic pressure increases
Greatly, the rupture of haemocyte can be accelerated, the dissolution rate of haemocyte is significantly improved, and cell protein will not be caused to be denatured；
Microwave radiation technology alkali enzyme is digested：Distilled water diluting is added into supernatant into 8-10g haemocytes/L, basic protein is added
Enzyme, makes enzyme concentration be 6000-7000U/g haemocytes, 50-60 DEG C of microwave radiation technology enzymolysis, wherein, plus dilute NaOH solution makes supernatant
PH value keep 9-10, obtain enzymolysis liquid；
Microwave radiation technology papain enzymolysis：Adding watery hydrochloric acid makes enzymolysis liquid pH value be 6-7, adds papain, makes
Enzyme concentration is 5000-6000U/g haemocytes, and 50-60 DEG C of microwave radiation technology is digested, and enzymolysis liquid is heated into 90-95 DEG C of inactivation,
Centrifugation, isolates supernatant；
After haemolysis, alkali enzyme, pawpaw egg enzyme enzymolysis are carried out successively, and both complete the enzyme of blood cell protein with combining harmony
Solution；Preferable stability is presented in enzymolysis process, the pH value of system, can effectively reduce because the cataclysm of pH environment influences albumen or polypeptide
The situation of denaturation；
Concentration：With 22 μm of microporous barriers by supernatant liquid filtering, Brix is concentrated under reduced pressure into for 50-60 °, deer hemalbumin polypeptide is produced
Paste liquid.
The deer hemalbumin polypeptide has higher oligopeptides ratio, it is easy to be absorbed by the body, while for preparing health beverages,
With preferable dispersiveness and stability.The paste liquid has the rheological characteristic of preferable stability, can further be spray-dried or
It is directly used in preparation health beverages.
Embodiment 1
A kind of deer hemalbumin polypeptide, its preparation method includes successively：
Washed corpuscles：Into 1L fresh deer bloods add 8g anti-coagulants, disperse, obtain anti-freezing blood sample, by anti-freezing blood sample with
3300rpm centrifuges 12min, discards blood plasma, collects haemocyte；In this step, anti-coagulants is citric acid, sodium citrate and glucose
By 1:(0.3-0.6):1 weight is than mixing；
Ultrasonic wave added haemolysis：By haemocyte brine, isometric distilled water is added, with 0.5mol/L's
NaOH regulations pH is 9-10, using 35-37 DEG C of ultrasonic wave added haemolysis 15min, ultrasonic power 350W, is centrifuged with 3300rpm
12min, collects supernatant；
Microwave radiation technology alkali enzyme is digested：Distilled water diluting is added into supernatant into 9g haemocytes/L, alkali protease is added,
Enzyme concentration is set to be 6500U/g haemocytes, 55 DEG C of microwave radiation technologies enzymolysis, wherein, plus 0.5mol/L NaOH solution makes supernatant
PH value keeps 9-10, obtains enzymolysis liquid；Microwave frequency is 2500MHz, and power is 5kW；
Microwave radiation technology papain enzymolysis：The watery hydrochloric acid for adding 0.5mol/L to enzymolysis liquid makes enzymolysis liquid pH value be 6-7,
Papain is added, it is 5000-6000U/g haemocytes to make enzyme concentration, 55 DEG C of microwave radiation technologies are digested, enzymolysis liquid is heated
To 95 rapid cooling DEG C inactivations, 12min is centrifuged with 3300rpm, supernatant is isolated；Microwave frequency is 2500MHz, and power is
5kW；
Concentration：With 22 μm of microporous barriers by supernatant liquid filtering, Brix is concentrated under reduced pressure into for 60 °, deer hemalbumin polypeptide is produced
Paste liquid.Counted on the basis of haemocyte, yield is 73.4%, oligopeptides ratio is 89.2%.
Embodiment 1
A kind of deer hemalbumin polypeptide, its preparation method includes successively：
Washed corpuscles：Into 1L fresh deer bloods add 8g anti-coagulants, disperse, obtain anti-freezing blood sample, by anti-freezing blood sample with
3300rpm centrifuges 12min, discards blood plasma, collects haemocyte；In this step, anti-coagulants is citric acid, sodium citrate and glucose
By 1:0.4:1 weight is than mixing；
Ultrasonic wave added haemolysis：By haemocyte brine, isometric distilled water is added, with 0.5mol/L's
NaOH regulations pH is 9-10, using 35-37 DEG C of ultrasonic wave added haemolysis 15min, ultrasonic power 300W, is centrifuged with 3300rpm
12min, collects supernatant；
Microwave radiation technology alkali enzyme is digested：Distilled water diluting is added into supernatant into 9g haemocytes/L, alkali protease is added,
Enzyme concentration is set to be 6500U/g haemocytes, 55 DEG C of microwave radiation technologies enzymolysis, wherein, plus 0.5mol/L NaOH solution makes supernatant
PH value keeps 9-10, obtains enzymolysis liquid；Microwave frequency is 2500MHz, and power is 5kW；
Microwave radiation technology papain enzymolysis：The watery hydrochloric acid for adding 0.5mol/L to enzymolysis liquid makes enzymolysis liquid pH value be 6-7,
Papain is added, it is 5500U/g haemocytes to make enzyme concentration, 55 DEG C of microwave radiation technologies are digested, and enzymolysis liquid is heated into 95
Rapid cooling DEG C inactivation, centrifuges 12min with 3300rpm, isolates supernatant；Microwave frequency is 2500MHz, and power is 5kW；
Concentration：With 22 μm of microporous barriers by supernatant liquid filtering, Brix is concentrated under reduced pressure into for 55 °, deer hemalbumin polypeptide is produced
Paste liquid.Counted on the basis of haemocyte, yield is 73.4%, oligopeptides ratio is 89.2%.
Embodiment 2
A kind of deer hemalbumin polypeptide, its preparation method includes successively：
Washed corpuscles：Into 1L fresh deer bloods add 8g anti-coagulants, disperse, obtain anti-freezing blood sample, by anti-freezing blood sample with
3300rpm centrifuges 12min, discards blood plasma, collects haemocyte；In this step, anti-coagulants is citric acid, sodium citrate and glucose
By 1:0.5:1 weight is than mixing；
Ultrasonic wave added haemolysis：By haemocyte brine, isometric distilled water is added, with 0.5mol/L's
NaOH regulations pH is 9-10, using 35-37 DEG C of ultrasonic wave added haemolysis 15min, ultrasonic power 400W, is centrifuged with 3500rpm
10min, collects supernatant；
Microwave radiation technology alkali enzyme is digested：Distilled water diluting is added into supernatant into 8g haemocytes/L, alkali protease is added,
Enzyme concentration is set to be 7000U/g haemocytes, 55 DEG C of microwave radiation technologies enzymolysis, wherein, plus 0.5mol/L NaOH solution makes supernatant
PH value keeps 9-10, obtains enzymolysis liquid；Microwave frequency is 2500MHz, and power is 5kW；
Microwave radiation technology papain enzymolysis：The watery hydrochloric acid for adding 0.5mol/L to enzymolysis liquid makes enzymolysis liquid pH value be 6-7,
Papain is added, it is 6000U/g haemocytes to make enzyme concentration, 55 DEG C of microwave radiation technologies are digested, and enzymolysis liquid is heated into 95
Rapid cooling DEG C inactivation, centrifuges 12min with 3300rpm, isolates supernatant；Microwave frequency is 2500MHz, and power is 5kW；
Concentration：With 22 μm of microporous barriers by supernatant liquid filtering, Brix is concentrated under reduced pressure into for 60 °, deer hemalbumin polypeptide is produced
Paste liquid.Counted on the basis of haemocyte, yield is 72.1%, oligopeptides ratio is 88.6%.
Embodiment 3
A kind of deer hemalbumin polypeptide, its preparation method includes successively：
Washed corpuscles：Into 1L fresh deer bloods add 8g anti-coagulants, disperse, obtain anti-freezing blood sample, by anti-freezing blood sample with
3000rpm centrifuges 15min, discards blood plasma, collects haemocyte；In this step, anti-coagulants is citric acid, sodium citrate and glucose
By 1:0.55:1 weight is than mixing；
Ultrasonic wave added haemolysis：By haemocyte brine, isometric distilled water is added, with 0.5mol/L's
NaOH regulations pH is 9-10, using 35-37 DEG C of ultrasonic wave added haemolysis 15min, ultrasonic power 350W, is centrifuged with 3500rpm
10min, collects supernatant；
Microwave radiation technology alkali enzyme is digested：Distilled water diluting is added into supernatant into 8g haemocytes/L, alkali protease is added,
Enzyme concentration is set to be 6000U/g haemocytes, 55 DEG C of microwave radiation technologies enzymolysis, wherein, plus 0.5mol/L NaOH solution makes supernatant
PH value keeps 9-10, obtains enzymolysis liquid；Microwave frequency is 2500MHz, and power is 5kW；
Microwave radiation technology papain enzymolysis：The watery hydrochloric acid for adding 0.5mol/L to enzymolysis liquid makes enzymolysis liquid pH value be 6-7,
Papain is added, it is 5000U/g haemocytes to make enzyme concentration, 55 DEG C of microwave radiation technologies are digested, and enzymolysis liquid is heated into 95
Rapid cooling DEG C inactivation, centrifuges 12min with 3300rpm, isolates supernatant；Microwave frequency is 2500MHz, and power is 5kW；
Concentration：With 22 μm of microporous barriers by supernatant liquid filtering, Brix is concentrated under reduced pressure into for 50 °, deer hemalbumin polypeptide is produced
Paste liquid.Counted on the basis of haemocyte, yield is 74.4%, oligopeptides ratio is 89.7%.
Comparative example 1：
A kind of deer hemalbumin polypeptide, its preparation method includes successively：
Washed corpuscles：Into 1L fresh deer bloods add 8g anti-coagulants, disperse, obtain anti-freezing blood sample, by anti-freezing blood sample with
3300rpm centrifuges 12min, discards blood plasma, collects haemocyte；In this step, anti-coagulants is citric acid, sodium citrate and glucose
By 1:0.4:1 weight is than mixing；
Haemolysis：By haemocyte brine, isometric distilled water is added, pH is adjusted with 0.5mol/L NaOH
For 9-10,35-37 DEG C of haemolysis 60min, 12min is centrifuged with 3300rpm, supernatant is collected；
Microwave radiation technology alkali enzyme is digested：Distilled water diluting is added into supernatant into 9g haemocytes/L, alkali protease is added,
Enzyme concentration is set to be 6500U/g haemocytes, 55 DEG C of microwave radiation technologies enzymolysis, wherein, plus 0.5mol/L NaOH solution makes supernatant
PH value keeps 9-10, obtains enzymolysis liquid；Microwave frequency is 2500MHz, and power is 5kW；
Microwave radiation technology papain enzymolysis：The watery hydrochloric acid for adding 0.5mol/L to enzymolysis liquid makes enzymolysis liquid pH value be 6-7,
Papain is added, it is 5500U/g haemocytes to make enzyme concentration, 55 DEG C of microwave radiation technologies are digested, and enzymolysis liquid is heated into 95
Rapid cooling DEG C inactivation, centrifuges 12min with 3300rpm, isolates supernatant；Microwave frequency is 2500MHz, and power is 5kW；
Concentration：With 22 μm of microporous barriers by supernatant liquid filtering, Brix is concentrated under reduced pressure into for 55 °, deer hemalbumin polypeptide is produced
Paste liquid.Counted on the basis of haemocyte, yield is 61.8%, oligopeptides ratio is 87.4%.
Comparative example 2：
A kind of deer hemalbumin polypeptide, its preparation method includes successively：
Washed corpuscles：Into 1L fresh deer bloods add 8g anti-coagulants, disperse, obtain anti-freezing blood sample, by anti-freezing blood sample with
3300rpm centrifuges 12min, discards blood plasma, collects haemocyte；In this step, anti-coagulants is citric acid, sodium citrate and glucose
By 1:0.4:1 weight is than mixing；
Ultrasonic wave added haemolysis：By haemocyte brine, isometric distilled water is added, with 0.5mol/L's
NaOH regulations pH is 9-10, using 35-37 DEG C of ultrasonic wave added haemolysis 15min, ultrasonic power 350W, is centrifuged with 3300rpm
12min, collects supernatant；
Alkali enzyme is digested：Distilled water diluting is added into supernatant into 9g haemocytes/L, alkali protease is added, makes enzyme concentration
For 6500U/g haemocytes, 55 DEG C of enzymolysis, wherein, plus 0.5mol/L NaOH solution makes the pH value of supernatant keep 9-10, obtains enzyme
Solve liquid；
Microwave radiation technology papain enzymolysis：The watery hydrochloric acid for adding 0.5mol/L to enzymolysis liquid makes enzymolysis liquid pH value be 6-7,
Papain is added, it is 5500U/g haemocytes to make enzyme concentration, 55 DEG C of microwave radiation technologies are digested, and enzymolysis liquid is heated into 95
Rapid cooling DEG C inactivation, centrifuges 12min with 3300rpm, isolates supernatant；Microwave frequency is 2500MHz, and power is 5kW；
Concentration：With 22 μm of microporous barriers by supernatant liquid filtering, Brix is concentrated under reduced pressure into for 55 °, deer hemalbumin polypeptide is produced
Paste liquid.Counted on the basis of haemocyte, yield is 73.6%, oligopeptides ratio is 60.1%.
Comparative example 3
A kind of deer hemalbumin polypeptide, its preparation method includes successively：
Washed corpuscles：Into 1L fresh deer bloods add 8g anti-coagulants, disperse, obtain anti-freezing blood sample, by anti-freezing blood sample with
3300rpm centrifuges 12min, discards blood plasma, collects haemocyte；In this step, anti-coagulants is citric acid, sodium citrate and glucose
By 1:(0.3-0.6):1 weight is than mixing；
Ultrasonic wave added haemolysis：By haemocyte brine, isometric distilled water is added, with 0.5mol/L's
NaOH regulations pH is 9-10, using 35-37 DEG C of ultrasonic wave added haemolysis 15min, ultrasonic power 350W, is centrifuged with 3300rpm
12min, collects supernatant；
Microwave radiation technology alkali enzyme is digested：Distilled water diluting is added into supernatant into 9g haemocytes/L, alkali protease is added,
Enzyme concentration is set to be 6500U/g haemocytes, 55 DEG C of microwave radiation technologies enzymolysis, wherein, plus 0.5mol/L NaOH solution makes supernatant
PH value keeps 9-10, obtains enzymolysis liquid；Microwave frequency is 2500MHz, and power is 5kW；
Papain enzymolysis：The watery hydrochloric acid for adding 0.5mol/L to enzymolysis liquid makes enzymolysis liquid pH value be 6-7, adds pawpaw
Protease, it is 5000-6000U/g haemocytes to make enzyme concentration, and 55 DEG C are digested, and enzymolysis liquid is heated into 95 rapid coolings DEG C goes out
It is living, 12min is centrifuged with 3300rpm, supernatant is isolated；
Concentration：With 22 μm of microporous barriers by supernatant liquid filtering, Brix is concentrated under reduced pressure into for 60 °, deer hemalbumin polypeptide is produced
Paste liquid.Counted on the basis of haemocyte, yield is 72.4%, oligopeptides ratio is 64.9%.
From embodiment 1-3 and comparative example 1-3 comparison, ultrasonic assistant haemolysis, yield are not used in the haemolysis stage
Substantially reduction；And crossed in alkali enzyme enzymolysis or papain enzymolysis and do not use microwave radiation technology means, although yield declines not clear
Show, but oligopeptides ratio is greatly reduced.
The deer hemalbumin polypeptide that the present invention is provided, has significant effect in terms of immune, anti-oxidant, can directly add
With obtained health beverages in wine product.
Performance detection and effect assessment
1st, determination oxidative
By 10 μm of ol/L Trolox storing solutions phosphorate phthalate buffer be diluted to concentration for 0.05,0.10,0.20,
0.40th, 0.80,1.6 μm of ol/L standard liquid, using phosphate buffer as negative control, using fluorescence half-life as vertical seat
Mark, using Trolox concentration as abscissa, draws standard curve.
Take 1g embodiment 1-3 deer hemalbumin polypeptide, plus 100mL phosphate buffer to be diluted, be well mixed,
Using distilled water as blank control, be respectively adopted phosphate buffer dilution by 10 times, 100 times, 1000 times, 10000 times, 100000
Dilute, dilution is measured, its result is as shown in the table again：
The oxidation resistance test result of table 1
2nd, mouse, which bears a heavy burden, tests
Using SPF male mices, body weight is 18-22g, per test group 10, totally 40, and swimming with a load attached to the body survey is carried out respectively
Fixed, environment temperature is 22-24 DEG C, humidity 50-54% during experiment.The deer blood egg polypeptide obtained using embodiment 1, setting dosage
For 0.1g/kg.bw, 0.2g/kg.bw, 0.5g/kg.bw, using aqua sterilisa as control group, mix food and feed 30 days, carry out swimming with a load attached to the body
Experiment.
The Loaned swimming test of table 2
After gavage is fed 30 days, each dosage group mouse, the walking weight load relative to control group mice is obviously prolonged.Say
The deer hemalbumin polypeptide that bright the application is provided, which has, improves body fatigue resistance effect.
3rd, stability test
By embodiment 1-3 and comparative example 1-3 deer hemalbumin polypeptide with 1:10 proportioning is added in 50 ° of white wine, concussion
Uniformly to precipitation is visible by naked eyes, the health preserving wine of deer hemalbumin polypeptide is obtained, 1L health preserving wines are taken with 3000rpm centrifugation
5min, observes health colours of wine and whether has precipitation, if so, precipitation volume is then measured, it is as a result as shown in the table：
The stability test result of table 3
As shown above, the deer hemalbumin polypeptide that this application is obtained has higher dynamic stability, the polypeptide
Prepare and be unlikely to deteriorate after health preserving wine, be not likely to produce precipitation, quality is higher.
Above-mentioned embodiment is only the preferred embodiment of the present invention, it is impossible to limit the scope of protection of the invention with this,
The change and replacement for any unsubstantiality that those skilled in the art is done on the basis of the present invention belong to institute of the present invention
Claimed scope.
Claims (10)
1. a kind of deer hemalbumin polypeptide, it is characterised in that its preparation method includes successively：
Washed corpuscles：Anti-coagulants is added into fresh deer blood, is disperseed, is obtained anti-freezing blood sample, anti-freezing blood sample is centrifuged, blood is discarded
Slurry, collects haemocyte；
Ultrasonic wave added haemolysis：By haemocyte brine, the distilled water of 1-2 times of volume is added, pH is adjusted with dilute NaOH
For 9-10, using 35-37 DEG C of ultrasonic wave added haemolysis, supernatant is collected in centrifugation；
Microwave radiation technology alkali enzyme is digested：Distilled water diluting is added into supernatant into 8-10g haemocytes/L, alkali protease is added,
Enzyme concentration is set to be 6000-7000U/g haemocytes, 50-60 DEG C of microwave radiation technology enzymolysis, wherein, plus dilute NaOH solution makes supernatant
PH value keeps 9-10, obtains enzymolysis liquid；
Microwave radiation technology papain enzymolysis：Adding watery hydrochloric acid makes enzymolysis liquid pH value be 6-7, adds papain, makes enzyme dense
Spend for 5000-6000U/g haemocytes, 50-60 DEG C of microwave radiation technology is digested, enzymolysis liquid is heated to 90-95 DEG C of inactivation, from
The heart, isolates supernatant；
Concentration：By supernatant liquid filtering, Brix is concentrated under reduced pressure into for 50-60 °, the paste liquid of deer hemalbumin polypeptide is produced.
2. deer hemalbumin polypeptide as claimed in claim 1, it is characterised in that in anti-freezing step, the anti-coagulants be citric acid,
The mixed solution of sodium citrate and glucose, wherein, the weight ratio of citric acid, sodium citrate and glucose is (0.8-1):
(0.3-0.6):1。
3. deer hemalbumin polypeptide as claimed in claim 1, it is characterised in that in washed corpuscles step, centrifugation rate is
3000-3500rpm, centrifugation time is 10-15min.
4. deer hemalbumin polypeptide as claimed in claim 1, it is characterised in that in ultrasonic wave added haemolysis step, ultrasonic power is
300-400W。
5. deer hemalbumin polypeptide as claimed in claim 1, it is characterised in that in microwave radiation technology alkali enzyme enzymolysis step, Microwave Frequency
Rate is 1500-3000MHz, and power is 5kW.
6. deer hemalbumin polypeptide as claimed in claim 1, it is characterised in that in microwave radiation technology alkali enzyme enzymolysis step, dilute NaOH
Concentration be 0.5mol/L.
7. deer hemalbumin polypeptide as claimed in claim 1, it is characterised in that in microwave radiation technology papain enzymolysis step,
Dilute HCl concentration is 0.5mol/L.
8. deer hemalbumin polypeptide as claimed in claim 1, it is characterised in that described to be filtered into micro porous filtration in concentration step.
9. deer hemalbumin polypeptide as claimed in claim 8, it is characterised in that the aperture of micro porous filtration is 22 μm.
10. application of the deer hemalbumin polypeptide on health products are prepared as described in claim any one of 1-9.
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| CN115466771A (en) * | 2022-10-12 | 2022-12-13 | 长春工业大学 | Preparation method and equipment of deer blood polypeptide chelated calcium |
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| CN115466771A (en) * | 2022-10-12 | 2022-12-13 | 长春工业大学 | Preparation method and equipment of deer blood polypeptide chelated calcium |
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