CN109105616A - A kind of method that scanning frequency pulse supersonic synergic alkali prepares walnut protein - Google Patents
A kind of method that scanning frequency pulse supersonic synergic alkali prepares walnut protein Download PDFInfo
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- CN109105616A CN109105616A CN201810993449.0A CN201810993449A CN109105616A CN 109105616 A CN109105616 A CN 109105616A CN 201810993449 A CN201810993449 A CN 201810993449A CN 109105616 A CN109105616 A CN 109105616A
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
- A23J1/146—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds by using wave energy or electric current
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Abstract
The invention discloses a kind of methods that scanning frequency pulse supersonic synergic alkali prepares walnut protein, belong to native protein extractive technique field.Specific steps include: that (1) utilizes oxygenation pretreatment walnut dregs raw material；(2) walnut protein extraction process is promoted using scanning frequency pulse ultrasonic treatment；(3) walnut protein is separated using acid for adjusting pH, walnut protein is made through centrifugation and vacuum freeze drying；The scanning frequency pulse supersonic synergic alkali that the present invention uses prepares walnut protein method, and operating procedure is simple and easy, and recovery rate is high, energy consumption is lower, has good market application prospect, can further increase the market economy benefit of petal resource.
Description
Technical field
The invention belongs to native protein extractive technique fields, and in particular to a kind of scanning frequency pulse supersonic synergic alkali prepares walnut
The method of albumen.
Background technique
China's walnut annual output is up to 2,710,000 tons, has exceeded the 50% of world's annual output, occupies world's umber one.Walnut
Nutrient content is more balanced, rich in the nutritional ingredients such as protein abundant, fat, belongs to comparatively ideal high protein, high-fat, and
And there is certain medical value.Walnut is mainly used as oil expression raw material and eats raw at present；Walnut dregs are the masters after squeezing walnut oil
Industrial by-products are wanted, main nutrient composition of the walnut kernel as this role of vegetable protein, protein are largely remained
Content is about 45%~54%, wherein the content of albumin, globulin, alcohol soluble protein and glutelin be respectively 6.8%,
17.6%, 5.3% and 70.1%.Walnut protein contains 18 kinds of amino acid, is rich in the necessary 8 kinds of amino acid of human body, especially paddy
Propylhomoserin and arginic content are very high；Therefore, walnut dregs still have very high use value and healthcare function, have good
Development prospect.However just from the point of view of current development and utilization situation, walnut dregs are mainly used as vegetable fertilizer and animal feed, and some adds
Work enterprise even directly discards, and causes the serious waste of walnut dregs.Add if the high-quality protein in walnut dregs can be extracted
With development and utilization, the economic value of petal resource will be greatly improved.
The conventional method of Protein Extraction is that alkali soluble acid sinks extraction method, but that there are alkali charges is big, recovery rate is low, consumption for this method
The deficiencies of duration.To solve the above-mentioned problems, research report mostly uses physics auxiliary extraction technology, such as Microwave-assisted Extraction at present
It follows the example of, supercritical fluid extraction, ultrasonic wave assisted extraction method etc..There is temperature during the extraction process and increase in microwave―assisted extraction
And it is difficult to the defect of temperature control, it is unfavorable for the reservation of thermal sensitivity walnut protein nutritive value；Although supercritical fluid extraction extracts
Rate is high, and easy temperature control system, but since high equipment cost haves the shortcomings that practical input cost is high.It is auxiliary compared to other physics
Extractive technique is helped, ultrasonic wave auxiliary extraction technology is more advantageous in terms of active material extraction, has mass-transfer efficiency height, extracts
The advantages such as the time is short, alkali charge is few, thermo-responsive compound destroys less, yield height, energy conservation and environmental protection.However, according to the text studied at present
From the point of view of offering report, there is not the extraction research that ultrasonic wave auxiliary extraction technology is used for albumen in walnut dregs also.
Summary of the invention
In view of the deficienciess of the prior art, the present invention introduces advanced sweep in the heavy extraction process of walnut protein alkali soluble acid
Frequency impulse ultrasound technology, can give full play to the synergistic effect of impulse ultrasound and frequency sweep ultrasonic, more efficiently promote albumen from
Walnut dregs raw material enters in aqueous slkali, to realize higher extraction efficiency.
The method that a kind of scanning frequency pulse supersonic synergic alkali of the present invention prepares walnut protein, as steps described below into
Row:
(1) walnut dregs are weighed to be added in distilled water, after stirring and evenly mixing, adjust pH；Then, it is placed in scanning frequency pulse ultrasonic device
In, setting ultrasonic treatment parameter carries out scanning frequency pulse ultrasonic extraction；After treatment is centrifuged, and supernatant is collected；
(2) supernatant obtained step (1) adjusts pH, after standing, is centrifuged, removes supernatant, collect precipitating, used
Distilled water cleans to arrive walnut protein extract；Protein extract obtained is subjected to vacuum freeze drying and obtains walnut
Albumen.
Preferably, the mass ratio of walnut dregs described in step (1) and distilled water is 1:12.
Preferably, adjusting pH described in step (1) is that material liquid pH value is adjusted with 4M sodium hydroxide to 9.0.
Preferably, ultrasonic treatment parameter described in step (1) are as follows: 15 DEG C of temperature, power density 133.3W/L, frequency 28
~68kHz；Frequency sweep 1.0~2.0kHz of amplitude, 20~1000ms of frequency sweep cycle；Pulse duty factor 33%~90%；Time 30~
150min。
Preferably, ultrasonic treatment parameter described in step (1) are as follows: 15 DEG C of temperature, power density 133.3W/L, frequency
28kHz；Frequency sweep amplitude 1.5kHz, frequency sweep cycle 100ms；Pulse duty factor 77%；Time 30min.
Preferably, adjusting pH described in step (2) is with 1M salt acid for adjusting pH value to 4.5.
Preferably, time of repose described in step (2) is 10~20min.
Preferably, centrifugal condition described in step (1) and (2) is 4000r/min, 15min.
Preferably, the condition of vacuum freeze drying described in step (2) are as follows: -40 DEG C of temperature, time 40h, vacuum degree -
0.08MPa。
The beneficial effects of the present invention are:
(1) the raw material walnut dregs that the present invention chooses are the by-products after walnut oil expression, are mainly used as vegetable fertilizer and animal
Feed, some are even directly dropped, and the utilization rate of walnut dregs is very low, causes the serious waste of walnut dregs resource；The present invention with
Walnut dregs are that raw material extracts high-quality walnut protein, help to improve the economic benefit of walnut dregs, expand the development of walnut dregs raw material
Range.
(2) present invention sinks in walnut protein alkali soluble acid introduces advanced scanning frequency pulse ultrasonic technique in extraction process, can be with
The synergistic effect of impulse ultrasound and frequency sweep ultrasonic is given full play to, more efficiently albumen is promoted to enter alkali soluble from walnut dregs raw material
In liquid, to realize higher extraction efficiency；On frequency sweep ultrasonic refers to that supersonic frequency is regular in a fixed range
Lower fluctuation can be carried out preferably matching generation resonant frequency with the intrinsic frequency of process object, reached compared with determining frequency ultrasound
Preferably dissolution effect；Impulse ultrasound refers to that ultrasound works time and intermittent time are alternately in extraction process, surpasses with continuous
Sound is compared, and the heating of reaction system is reduced, more energy saving.
(3) the scanning frequency pulse supersonic synergic alkali extracting method that the present invention uses, at low cost, device simple, operating procedure are not
Cumbersome and recovery rate is high, helps to apply in industrialized production, in technology and equipment commercialization, standardization, marketization side
Mask has great advantage.
Detailed description of the invention
Fig. 1 is scanning frequency pulse ultrasonic device structure chart.
The reference numerals are as follows:
The upper ultrasonic transducer height adjusting screw rod of 1-, the upper ultrasonic transducer of 2-, ultrasonic transducer under 3-, 4- ultrasound pond, 5-
Water outlet, 6- computer controller, 7- ultrasonic wave controller.
Specific embodiment:
The present invention is described in further detail below with reference to specific embodiment, and referring to data.The embodiment of the present invention
Technical solution of the present invention solely for the purpose of illustration, rather than limit the scope of the invention in any way.
Fig. 1 is the scanning frequency pulse ultrasound processing equipment that the present invention uses, and is Jiangsu University's independent development.The equipment is furnished with one
Platform computer controller 6 can set ultrasound works parameter (frequency, frequency sweep amplitude, frequency sweep cycle, pulse duty factor, time), so
Ultrasonic wave controller 7 is controlled afterwards, and scanning frequency pulse ultrasonication is realized by upper and lower ultrasonic transducer 2 and 3；After alkali process
Walnut feed liquid be placed in ultrasonic pond 4, carry out different ultrasonic treatments and test；By adjusting upper ultrasonic transducer height adjusting screw rod
The sample-loading amount in 1 height and ultrasonic pond 4 is come to control ultrasonic power density be 133.3W/L.
Embodiment 1:
Not sonicated walnut protein preparation method carries out as steps described below:
(1) walnut dregs and distilled water are weighed by solid-liquid ratio 1:12, after mixing evenly, adjusts material liquid pH value with 4M sodium hydroxide
To 9.0；
(2) feed liquid that step (1) obtains is stirred into 30min at 15 DEG C of bath temperature, then by feed liquid in 4000r/min
It is centrifuged 15min under revolving speed, collects supernatant；
(3) supernatant for obtaining step (2) 1M hydrochloric acid adjusts material liquid pH value to 4.5, after standing, in 4000r/min
It is centrifuged 15min under revolving speed, removes supernatant, collects precipitating, 2 times are to obtain walnut protein extract wash with distilled water；
(4) by protein extract vacuum freeze drying made from step (3), the condition of vacuum freeze drying are as follows: temperature -40
DEG C, time 40h, vacuum degree -0.08MPa, obtain walnut protein；
(5) walnut protein yield and assay:
The measurement of walnut protein yield: 5g Walnut protein powder accurately is weighed, is placed in a beaker, distilled water is added to dissolve and adjust
PH value of solution is to 11.0, after albumen is completely dissolved, is settled in the volumetric flask of 500mL, after then sample is suitably diluted according to
Forint phenol method measures sample protein concentration.
Walnut protein yield calculation formula:
In formula: the protein concentration (mg/mL) that c-is pushed away according to bovine serum albumin(BSA) standard curve；Used in v-constant volume albumen
Volumetric flask specification (500mL)；m0- weighed albumen quality (5g)；N-sample extension rate；M-acquisition albumen is total
Quality (g)；M-walnut dregs raw material quality (g).
Walnut protein content calculation formula:
The concrete outcome of walnut protein yield and content is shown in Table 1.
Embodiment 2:
(1) walnut dregs and distilled water are weighed by solid-liquid ratio 1:12, after mixing evenly, adjusts material liquid pH value with 4M sodium hydroxide
To 9.0；
(2) feed liquid that step (1) obtains is poured into frequency sweep ultrasonic equipment and carries out scanning frequency pulse ultrasonic extraction, ultrasonic treatment ginseng
Number are as follows: 15 DEG C of temperature；Power density 133.3W/L；Frequency 28kHz；Frequency sweep amplitude 2.0kHz；Frequency sweep cycle 100ms；Pulse accounts for
Empty ratio 77%；Time 30min；After ultrasonic treatment, feed liquid is centrifuged 15min under 4000r/min revolving speed, collects supernatant
Liquid；
(3) supernatant for obtaining step (2) 1M hydrochloric acid adjusts material liquid pH value to 4.5, after standing, in 4000r/min
It is centrifuged 15min under revolving speed, removes supernatant, collects precipitating, 2 times are to obtain walnut protein extract wash with distilled water；
(4) by protein extract vacuum freeze drying made from step (3), the condition of vacuum freeze drying are as follows: temperature -40
DEG C, time 40h, vacuum degree -0.08MPa, obtain walnut protein；
(5) walnut protein yield and assay:
The measurement of walnut protein yield: 5g Walnut protein powder accurately is weighed, is placed in a beaker, distilled water is added to dissolve and adjust
PH value of solution is to 11.0, after albumen is completely dissolved, is settled in the volumetric flask of 500mL, after then sample is suitably diluted according to
Forint phenol method measures sample protein concentration.
Walnut protein yield calculation formula:
In formula: the protein concentration (mg/mL) that c-is pushed away according to bovine serum albumin(BSA) standard curve；Used in v-constant volume albumen
Volumetric flask specification (500mL)；m0- weighed albumen quality (5g)；N-sample extension rate；M-acquisition albumen is total
Quality (g)；M-walnut dregs raw material quality (g).
Walnut protein content calculation formula:
The concrete outcome of walnut protein yield and content is shown in Table 1.
Embodiment 3:
The frequency of ultrasonic treatment is 40kHz, remaining step is the same as embodiment 2；The concrete outcome of walnut protein yield and content
It is shown in Table 1.
Embodiment 4:
The frequency of ultrasonic treatment is 68kHz, remaining step is the same as embodiment 2；The concrete outcome of walnut protein yield and content
It is shown in Table 1.
Influence of the different supersonic frequencies of table 1 to walnut protein yield and content
By comparative example 1 in table 1 with the walnut protein yield and content of embodiment 2-4 it can be found that with compareing (no
Add ultrasound) it compares, the ultrasonic treatment of different frequency can make walnut protein yield increase, especially supersonic frequency 28kHz's
Ultrasonic treatment makes walnut protein recovery rate improve 1.4%, while the content of walnut protein being made to improve 1.0%.Comprehensively consider
From the point of view of walnut protein yield and content, moderate ultrasonic frequency is conducive to the extraction of albumen, when wherein frequency is 28kHz, core
Peach protein extraction effect is optimal.
Embodiment 5:
(1) walnut dregs and distilled water are weighed by solid-liquid ratio 1:12, after mixing evenly, adjusts material liquid pH value with 4M sodium hydroxide
To 9.0；
(2) feed liquid that step (1) obtains is poured into frequency sweep ultrasonic equipment and carries out scanning frequency pulse ultrasonic extraction, ultrasonic treatment ginseng
Number are as follows: 15 DEG C of temperature；Power density 133.3W/L；Frequency 28kHz；Frequency sweep amplitude 1.0kHz；Frequency sweep cycle 100ms；Pulse accounts for
Empty ratio 77%；Time 30min.After ultrasonic treatment, feed liquid is centrifuged 15min under 4000r/min revolving speed, collects supernatant
Liquid；
(3) supernatant for obtaining step (2) 1M hydrochloric acid adjusts material liquid pH value to 4.5, after standing, in 4000r/min
It is centrifuged 15min under revolving speed, removes supernatant, collects precipitating, 2 times are to obtain walnut protein extract wash with distilled water；
(4) protein extract made from step (3) is subjected to vacuum freeze drying, the condition of vacuum freeze drying are as follows: temperature
- 40 DEG C, time 40h, vacuum degree -0.08MPa are spent, walnut protein is obtained；
(5) walnut protein yield and assay:
The measurement of walnut protein yield: 5g Walnut protein powder accurately is weighed, is placed in a beaker, distilled water is added to dissolve and adjust
PH value of solution is to 11.0, after albumen is completely dissolved, is settled in the volumetric flask of 500mL, after then sample is suitably diluted according to
Forint phenol method measures sample protein concentration.
Walnut protein yield calculation formula:
In formula: the protein concentration (mg/mL) that c-is pushed away according to bovine serum albumin(BSA) standard curve；Used in v-constant volume albumen
Volumetric flask specification (500mL)；m0- weighed albumen quality (5g)；N-sample extension rate；M-acquisition albumen is total
Quality (g)；M-walnut dregs raw material quality (g).
Walnut protein content calculation formula:
The concrete outcome of walnut protein yield and content is shown in Table 2.
Embodiment 6:
The frequency sweep amplitude of ultrasonic treatment is 1.5kHz, remaining step is the same as embodiment 5；Walnut protein yield and content it is specific
It the results are shown in Table 2.
Embodiment 7:
The frequency sweep amplitude of ultrasonic treatment is 2.0kHz, remaining step is the same as embodiment 5；Walnut protein yield and content it is specific
It the results are shown in Table 2.
Influence of the different frequency sweep amplitudes of table 2 to walnut protein yield and content
| Embodiment | Frequency sweep amplitude (kHz) | Yield (%) | Content (%) |
| Embodiment 1 (comparative example) | Ultrasound is not added | 8.94 | 31.63 |
| Embodiment 5 | 1.0 | 12.51 | 30.08 |
| Embodiment 6 | 1.5 | 14.73 | 34.74 |
| Embodiment 7 | 2.0 | 10.34 | 32.63 |
By comparative example 1 in table 2 with the walnut protein yield and content of embodiment 5-7 it can be found that with compareing (no
Add ultrasound) it compares, the ultrasonic treatment of different frequency sweep amplitudes can be such that walnut protein yield improves, and 1.4%- has been respectively increased
5.79%, especially frequency sweep amplitude are that the ultrasonic treatment of 1.0kHz and 1.5kHz makes walnut protein yield improve 3.57% respectively
With 5.79%.The ultrasonic treatment that frequency sweep amplitude is 1.5kHz also improves walnut protein content simultaneously, improves 3.11%.It is comprehensive
Consider walnut protein yield and content, frequency sweep amplitude is that the walnut protein that the scanning frequency pulse of 1.5kHz is ultrasonically treated extracts effect
Fruit is optimal.
Embodiment 8:
(1) walnut dregs and distilled water are weighed by solid-liquid ratio 1:12, after mixing evenly, adjusts material liquid pH value with 4M sodium hydroxide
To 9.0；
(2) feed liquid that step (1) obtains is poured into frequency sweep ultrasonic equipment and carries out scanning frequency pulse ultrasonic extraction, ultrasonic treatment ginseng
Number are as follows: 15 DEG C of temperature；Power density 133.3W/L；Frequency 28kHz；Frequency sweep amplitude 1.5kHz；Frequency sweep cycle 20ms；Pulse duration
Than 77%；Time 30min.After ultrasonic treatment, feed liquid is put into rapidly centrifuge, is centrifuged under 4000r/min revolving speed
15min collects supernatant；
(3) supernatant for obtaining step (2) 1M hydrochloric acid adjusts material liquid pH value to 4.5, after standing, in 4000r/min
It is centrifuged 15min under revolving speed, removes supernatant, collects precipitating, 2 times are to obtain walnut protein extract wash with distilled water；
(4) protein extract made from step (3) is subjected to vacuum freeze drying, the condition of vacuum freeze drying are as follows: temperature
- 40 DEG C, time 40h, vacuum degree -0.08MPa are spent, walnut protein is obtained；
(5) walnut protein yield and assay:
The measurement of walnut protein yield: 5g Walnut protein powder accurately is weighed, is placed in a beaker, distilled water is added to dissolve and adjust
PH value of solution is to 11.0, after albumen is completely dissolved, is settled in the volumetric flask of 500mL, after then sample is suitably diluted according to
Forint phenol method measures sample protein concentration.
Walnut protein yield calculation formula:
In formula: the protein concentration (mg/mL) that c-is pushed away according to bovine serum albumin(BSA) standard curve；Used in v-constant volume albumen
Volumetric flask specification (500mL)；m0- weighed albumen quality (5g)；N-sample extension rate；M-acquisition albumen is total
Quality (g)；M-walnut dregs raw material quality (g).
Walnut protein content calculation formula:
The concrete outcome of walnut protein yield and content is shown in Table 3.
Embodiment 9:
The frequency sweep cycle of ultrasonic treatment is 100ms, remaining step is the same as embodiment 8；Walnut protein yield and content it is specific
It the results are shown in Table 3.
Embodiment 10:
The frequency sweep cycle of ultrasonic treatment is 1000ms, remaining step is the same as embodiment 8；Walnut protein yield and content it is specific
It the results are shown in Table 3.
Influence of the different frequency sweep cycles of table 3 to walnut protein yield and content
| Embodiment | Frequency sweep cycle (ms) | Yield (%) | Content (%) |
| Embodiment 1 (comparative example) | Ultrasound is not added | 8.94 | 31.63 |
| Embodiment 8 | 20 | 10.10 | 28.98 |
| Embodiment 9 | 100 | 14.73 | 34.74 |
| Embodiment 10 | 1000 | 13.62 | 29.52 |
By the walnut protein yield and content of comparative example 1 in table 3 and embodiment 8-10 it can be found that with to photograph
Than the ultrasonic treatment of different frequency sweep cycles can make walnut protein yield improve 1.16%-5.79%, especially frequency sweep cycle
The ultrasonic treatment of 100ms and 1000ms makes walnut protein yield improve 5.79% and 4.68% respectively.Frequency sweep cycle 100ms's is super
Sonication also improves walnut protein content simultaneously, improves 3.11%.Comprehensively consider walnut protein yield and content, frequency sweep week
Phase is that the walnut protein extraction effect that the scanning frequency pulse of 100ms is ultrasonically treated is optimal.
Embodiment 11:
(1) walnut dregs and distilled water are weighed by solid-liquid ratio 1:12, after mixing evenly, adjusts material liquid pH value with 4M sodium hydroxide
To 9.0；
(2) feed liquid that step (1) obtains is poured into frequency sweep ultrasonic equipment and carries out scanning frequency pulse ultrasonic extraction, ultrasonic treatment ginseng
Number are as follows: 15 DEG C of temperature；Power density 133.3W/L；Frequency 28kHz；Frequency sweep amplitude 1.5kHz；Frequency sweep cycle 100ms；Pulse accounts for
33% (being specifically shown in Table 4) of empty ratio；Time 30min.After ultrasonic treatment, feed liquid is centrifuged under 4000r/min revolving speed
15min collects supernatant；
(3) supernatant for obtaining step (2) 1M hydrochloric acid adjusts material liquid pH value to 4.5, after standing, in 4000r/min
It is centrifuged 15min under revolving speed, removes supernatant, collects precipitating, 2 times are to obtain walnut protein extract wash with distilled water；
(4) protein extract made from step (3) is subjected to vacuum freeze drying, the condition of vacuum freeze drying are as follows: temperature
- 40 DEG C, time 40h, vacuum degree -0.08MPa are spent, walnut protein is obtained；
(5) walnut protein yield and assay:
The measurement of walnut protein yield: 5g Walnut protein powder accurately is weighed, is placed in a beaker, distilled water is added to dissolve and adjust
PH value of solution is to 11.0, after albumen is completely dissolved, is settled in the volumetric flask of 500mL, after then sample is suitably diluted according to
Forint phenol method measures sample protein concentration.
Walnut protein yield calculation formula:
In formula: the protein concentration (mg/mL) that c-is pushed away according to bovine serum albumin(BSA) standard curve；Used in v-constant volume albumen
Volumetric flask specification (500mL)；m0- weighed albumen quality (5g)；N-sample extension rate；M-acquisition albumen is total
Quality (g)；M-walnut dregs raw material quality (g).
Walnut protein content calculation formula:
The concrete outcome of walnut protein yield and content is shown in Table 4.
Embodiment 12:
The pulse duty factor of ultrasonic treatment is 77%, remaining step is the same as embodiment 11.The tool of walnut protein yield and content
Body the results are shown in Table 4.
Embodiment 13:
The pulse duty factor of ultrasonic treatment is 90%, remaining step is the same as embodiment 11.The tool of walnut protein yield and content
Body the results are shown in Table 4.
The different pulse durations of table 4 compare the influence of walnut protein yield and content
By comparative example 1 in table 4 with the walnut protein yield and content of embodiment 11-13 it can be found that with compareing
It compares, the ultrasonic treatment of different pulse duty factors can make walnut protein yield improve 0.82%-5.79%, and especially ultrasound accounts for
Sky than making walnut protein yield improve 5.79% and 2.38% respectively for 77% and 90% ultrasonic treatment.Same discovery,
Compared with the control, the ultrasonic treatment that ultrasound duty cycle is 77% and 90% makes walnut protein content improve 3.11% He respectively
0.1%, it is most that the ultrasonic treatment that wherein duty ratio is 77% improves walnut protein content, reaches 3.11%.Comprehensively consider
Walnut protein yield and content, the walnut protein extraction effect that the scanning frequency pulse that pulse duty factor is 77% is ultrasonically treated is most
It is excellent.
Embodiment 14:
(1) walnut dregs and distilled water are weighed by solid-liquid ratio 1:12, after mixing evenly, adjusts material liquid pH value with 4M sodium hydroxide
To 9.0；
(2) feed liquid that step (1) obtains is poured into frequency sweep ultrasonic equipment and carries out scanning frequency pulse ultrasonic extraction, ultrasonic treatment ginseng
Number are as follows: 15 DEG C of temperature；Power density 133.3W/L；Frequency 28kHz；Frequency sweep amplitude 1.5kHz；Frequency sweep cycle 100ms；Pulse accounts for
Empty ratio 77%；Time 30min；After ultrasonic treatment, feed liquid is centrifuged 15min under 4000r/min revolving speed, collects supernatant
Liquid；
(3) supernatant for obtaining step (2) 1M hydrochloric acid adjusts material liquid pH value to 4.5, after standing, in 4000r/min
It is centrifuged 15min under revolving speed, removes supernatant, collects precipitating, 2 times are to obtain walnut protein extract wash with distilled water；
(4) protein extract made from step (3) is subjected to vacuum freeze drying, the condition of vacuum freeze drying are as follows: temperature
- 40 DEG C, time 40h, vacuum degree -0.08MPa are spent, walnut protein is obtained；
(5) walnut protein yield and assay:
The measurement of walnut protein yield: 5g Walnut protein powder accurately is weighed, is placed in a beaker, distilled water is added to dissolve and adjust
PH value of solution is to 11.0, after albumen is completely dissolved, is settled in the volumetric flask of 500mL, after then sample is suitably diluted according to
Forint phenol method measures sample protein concentration.
Walnut protein yield calculation formula:
In formula: the protein concentration (mg/mL) that c-is pushed away according to bovine serum albumin(BSA) standard curve；Used in v-constant volume albumen
Volumetric flask specification (500mL)；m0- weighed albumen quality (5g)；N-sample extension rate；M-acquisition albumen is total
Quality (g)；M-walnut dregs raw material quality (g)；
Walnut protein content calculation formula:
The concrete outcome of walnut protein yield and content is shown in Table 5.
Embodiment 15:
The time of ultrasonic treatment is 90min, remaining step is the same as embodiment 14；The concrete outcome of walnut protein yield and content
It is shown in Table 5.
Embodiment 16:
The time of ultrasonic treatment is 150min, remaining step is the same as embodiment 14；The specific knot of walnut protein yield and content
Fruit is shown in Table 5.
Influence of the different ultrasonic times of table 5 to walnut protein yield and content
| Embodiment | Time (min) | Yield (%) | Content (%) |
| Embodiment 1 (comparative example) | Ultrasound is not added | 8.94 | 31.63 |
| Embodiment 14 | 30 | 14.73 | 34.74 |
| Embodiment 15 | 90 | 14.30 | 32.27 |
| Embodiment 16 | 150 | 13.84 | 31.20 |
By comparative example 1 in table 5 with the walnut protein yield and content of embodiment 14-16 it can be found that with compareing
It compares, the ultrasonic treatment of different time can make walnut protein yield improve 4.9%-5.79%, especially ultrasonic time 30min
Ultrasonic treatment makes walnut protein yield and content improve highest.Comprehensively consider walnut protein yield and content, ultrasonic time is
The walnut protein extraction effect that the scanning frequency pulse of 30min is ultrasonically treated is optimal.
Obtained according to above-mentioned experimental result, in embodiment 14 scanning frequency pulse supersonic synergic alkali carries take walnut protein be not added it is super
The embodiment 1 (control group) of sound is compared, and yield improves 5.79%, and content improves 3.11%, it follows that, scanning frequency pulse is super
Sonication walnut protein extraction effect is substantially better than the walnut protein extraction effect not being ultrasonically treated.
Claims (9)
1. a kind of method that scanning frequency pulse supersonic synergic alkali prepares walnut protein, which is characterized in that steps are as follows:
(1) walnut dregs are weighed to be added in distilled water, after stirring and evenly mixing, adjust pH；Then, it is placed in scanning frequency pulse ultrasonic device,
Setting ultrasonic treatment parameter, carries out scanning frequency pulse ultrasonic extraction；After treatment is centrifuged, and supernatant is collected；
(2) supernatant obtained step (1) adjusts pH, after standing, is centrifuged, removes supernatant, precipitating is collected, with distillation
Water cleans to arrive walnut protein extract；Protein extract obtained is subjected to vacuum freeze drying, i.e. acquisition walnut egg
It is white.
2. the method that a kind of scanning frequency pulse supersonic synergic alkali according to claim 1 prepares walnut protein, which is characterized in that
The mass ratio of walnut dregs described in step (1) and distilled water is 1:12.
3. the method that a kind of scanning frequency pulse supersonic synergic alkali according to claim 1 prepares walnut protein, feature exist
In it is 9.0 that adjusting pH described in step (1), which is with 4 M sodium hydroxides adjusting pH value,.
4. the method that a kind of scanning frequency pulse supersonic synergic alkali according to claim 1 prepares walnut protein, feature exist
In ultrasonic treatment parameter as described in step (1) are as follows: 15 DEG C of temperature, 133.3 W/L of power density, 28 ~ 68 kHz of frequency；It sweeps
1.0 ~ 2.0 kHz of frequency vibration；20 ~ 1000 ms of frequency sweep cycle；Pulse duty factor 33% ~ 90%；30 ~ 150 min of time.
5. the method that a kind of scanning frequency pulse supersonic synergic alkali according to claim 4 prepares walnut protein, feature exist
In the ultrasonic treatment parameter are as follows: 15 DEG C of temperature, 133.3 W/L of power density, 28 kHz of frequency；Frequency sweep amplitude 1.5
kHz；100 ms of frequency sweep cycle；Pulse duty factor 77%；30 min of time.
6. the method that a kind of scanning frequency pulse supersonic synergic alkali according to claim 1 prepares walnut protein, feature exist
In it is 4.5 that adjusting pH described in step (2), which is with 1 M salt acid for adjusting pH value,.
7. the method that a kind of scanning frequency pulse supersonic synergic alkali according to claim 1 prepares walnut protein, feature exist
In time of repose described in step (2) is 10 ~ 20 min.
8. the method that a kind of scanning frequency pulse supersonic synergic alkali according to claim 1 prepares walnut protein, feature exist
In centrifugal condition described in step (1) and (2) is equal are as follows: 4000 r/min of revolving speed, 15 min of time.
9. the method that a kind of scanning frequency pulse supersonic synergic alkali according to claim 1 prepares walnut protein, feature exist
In the condition of vacuum freeze drying described in step (2) are as follows: -40 DEG C of temperature, 40 h of time, -0.08 MPa of vacuum degree.
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