CN115651954A - Protein zymolyte and preparation and application thereof - Google Patents
Protein zymolyte and preparation and application thereof Download PDFInfo
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- CN115651954A CN115651954A CN202211378533.4A CN202211378533A CN115651954A CN 115651954 A CN115651954 A CN 115651954A CN 202211378533 A CN202211378533 A CN 202211378533A CN 115651954 A CN115651954 A CN 115651954A
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Abstract
The invention provides a protein zymolyte and preparation and application thereof. The preparation method of the protein zymolyte comprises the following steps: pulverizing animals and plants, adding water, sequentially performing high pressure treatment, enzymolysis, and low temperature plasma treatment; wherein the pressure of the high-pressure treatment is 100-400 MPa. According to the invention, the salty taste of the seasoning can be remarkably improved by the obtained proteolysis product through the high-pressure synergistic low-temperature plasma treatment (the salty taste intensity of 50mmol/L NaCl solution can be increased by 45.6% -57.2% by the proteolysis product with the protein content of 20 mg/mL), and the proteolysis product can be used as a high-quality salty taste enhancer and is beneficial to guaranteeing the dietary health of consumers.
Description
Technical Field
The invention belongs to the technical field of food processing. More particularly, it relates to a proteolytic enzyme, its preparation and application.
Background
Sodium salt, which is the most widely used seasoning at present, plays an important role in the daily life of people, and can impart salty taste to food and improve the overall flavor, but excessive intake of sodium salt by consumers can increase the incidence of hypertension and cardiovascular diseases. Therefore, sodium salt substitutes gradually enter the lives of consumers, for example, metal salts such as potassium chloride, magnesium chloride and the like which are commonly used at present can reduce the intake of sodium salts by the consumers, but the metal salts are bitter and astringent and are difficult to be accepted by the consumers, and potassium ions are not suitable for the consumers suffering from diseases such as renal insufficiency, hyperkalemia and the like. Therefore, the development of new low-sodium seasoning with flavor similar to sodium salt is becoming a new goal of researchers, such as searching for a seasoning additive capable of enhancing salty taste, and reducing Na + The salty taste intensity of the seasoning can be still ensured under the condition of content, the food is endowed with salty taste, the integral flavor of the food is improved, and the method is a good method for guaranteeing the diet health of consumers.
After the animal and plant proteins are subjected to enzymolysis by protease, small molecular peptides and free amino acids can be released, different process treatments are beneficial to releasing more small molecular peptides and free amino acids, the characteristics of zymolyte are improved, the zymolyte has richer and full-bodied flavor, products which can be used as seasoning additives are not lacked, but the zymolyte obtained by directly carrying out enzymolysis on the proteins has limited effect of improving the salty taste of the seasoning, and cannot become a high-quality salty taste enhancer.
The new article of poplar et al discloses that low temperature plasma treatment can chemically change proteins in food systems (new articles of poplar, niuwenjun, juniper, et al. Low temperature plasma technology and its effect on food quality and microorganisms [ J ] food and machinery, 2019,35 (9): 199-203,215. Doi.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a preparation method of a protein zymolyte, which utilizes high pressure to cooperate with low temperature plasma treatment to obviously improve the salty taste enhancement effect of the protein zymolyte, further effectively improve the salty taste of a seasoning, is a high-quality salty taste enhancer and is beneficial to ensuring the dietary health of consumers.
Another object of the present invention is to provide a proteolytic enzyme prepared by the above method.
Still another object of the present invention is to provide the use of the above-mentioned proteolytic hydrolysate as a salty taste enhancer and/or for the preparation of a salty taste enhancer.
The above purpose of the invention is realized by the following technical scheme:
the invention provides a preparation method of a protein zymolyte, which comprises the following steps: crushing animals and plants, adding water, and sequentially performing high-pressure treatment, enzymolysis and low-temperature plasma treatment to obtain the protein zymolyte;
wherein the pressure of the high-pressure treatment is 100-400 MPa.
Preferably, the animal and plant is an animal and plant with a protein content of more than 15%.
Preferably, the low-temperature plasma treatment is: low-temperature plasma treatment is carried out for 5-15 min at the power of 30-40 w and the temperature of-2 ℃. Most preferably, the low temperature plasma treatment is performed at a power of 40w and a temperature of 0 ℃ for 5min.
Preferably, the generating gas of the low-temperature plasma treatment is air.
Further preferably, the flow rate of the gas is 3 to 5L/min. Most preferably 5L/min.
Preferably, the mass ratio of the water to the crushed animals and plants is 1.8-2.2: 1. most preferably 2:1.
preferably, the high pressure treatment time is 15 to 25min. Most preferably 20min.
Preferably, the temperature of the high pressure treatment is 20 to 30 ℃. Most preferably 25 deg.c.
Preferably, the enzyme used for the enzymatic hydrolysis is a protease.
Further preferably, the enzyme comprises one or more of papain, trypsin or flavourzyme.
More preferably, the mass of the enzyme is 1.8-2.2 per mill of the mass of the animals and plants after being crushed. Most preferably 2% o.
Preferably, the enzymatic hydrolysis is: enzymolysis is carried out for 3.5-4.5 h at the temperature of 45-55 ℃. Most preferably at 50 ℃ for 4h.
Preferably, after the enzymolysis, enzyme deactivation is carried out by boiling for 15-25 min. Most preferably 20min.
Further preferably, centrifugation is performed after the enzyme inactivation.
More preferably, the centrifugation is carried out at 4500-5000 g and 3-5 deg.C for 18-22 min. Most preferably, centrifugation is carried out at 4800g and 4 ℃ for 20min.
Most preferably, the centrifugation is followed by suction filtration.
Preferably, the animals and plants are one or more of culled laying hens, breams or peas.
Preferably, when the animal and plant is culled layer chicken, the crushing is as follows: removing skin, bone and fat of obsolete laying hens, cutting into small pieces, and mincing into meat paste.
The proteolysis product obtained by utilizing high pressure and low temperature plasma treatment has excellent salty taste enhancement effect, can effectively enhance the salty taste of the seasoning, is a high-quality salty taste enhancer and is beneficial to guaranteeing the diet health of consumers, so the proteolysis product prepared by the method and the application of the proteolysis product as and/or in preparing the salty taste enhancer are also within the protection range of the invention.
The invention has the following beneficial effects:
1. according to the invention, through high-pressure synergistic low-temperature plasma treatment, the obtained protein zymolyte can remarkably improve the salty taste (the protein zymolyte with the protein content of 20mg/mL can increase the salty taste intensity of 50mmol/L NaCl solution by 45.6% -57.2%) and the delicate flavor of the seasoning, can be used as a high-quality salty taste enhancer and delicate flavor enhancer, and is beneficial to guaranteeing the dietary health of consumers.
2. The protein zymolyte of the invention has simple preparation, easily obtained raw materials, low cost, high nutritive value, easy absorption and utilization by human body, and has the functions of self oxidation resistance, bacteriostasis and the like of the small molecular peptide, and contains a large amount of small molecular polypeptide.
Drawings
FIG. 1 shows the results of an electronic tongue test.
FIG. 2 is a statistical graph of peptide molecular weight distribution.
Detailed Description
The invention is further described with reference to the drawings and specific examples, which are not intended to limit the invention in any way. The reagents, methods and apparatus employed in the present invention are conventional in the art, except as otherwise indicated.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
EXAMPLE 1 preparation of the proteolytic digest
S1, unfreezing frozen rejected laying hens (purchased from Guangdong Wenshi food group, ltd.) in a refrigerator at 4 ℃ for 12 hours, peeling, deboning and degreasing the unfrozen rejected laying hens, cutting the rejected laying hens into small pieces, and mincing the small pieces by a meat mincer with the diameter of a pore plate being 6mm to obtain chicken meat paste;
s2, taking 20g of chicken meat paste, adding 40g of deionized water, uniformly mixing, packaging in a vacuum packaging bag, and placing in a high-pressure device for high-pressure treatment for 20min at the temperature of 25 ℃ and under the pressure of 100 MPa;
s3, after high-pressure treatment, adding papain (the mass of the papain is 2 per mill of the mass of the minced meat), carrying out enzymolysis for 4 hours in a water-bath constant-temperature oscillator at 50 ℃, boiling for 20min to inactivate enzyme, cooling to 25 ℃, centrifuging for 20min at 4800g and 4 ℃, and carrying out suction filtration to obtain an enzymolysis product;
s4, placing the enzymolysis product obtained in the step S3 in a low-temperature plasma device, and carrying out low-temperature plasma treatment for 15min at the power of 30w and the temperature of 0 ℃, wherein the generated gas is air, and the gas flow rate is 3L/min.
EXAMPLE 2 preparation of the proteolytic digest
The same as example 1 except that the pressure of the high-pressure treatment was 200MPa; the low-temperature plasma treatment comprises the following steps: the low-temperature plasma treatment is carried out for 5min at the power of 40w and the temperature of 0 ℃, the generated gas is air, and the gas flow rate is 5L/min.
EXAMPLE 3 preparation of the proteolytic digest
The difference from example 1 is that the low temperature plasma treatment is: the low-temperature plasma treatment is carried out for 10min at the power of 40w and the temperature of 0 ℃, the generated gas is air, and the gas flow rate is 4L/min.
Comparative example 1
The difference from example 1 is that the low temperature plasma treatment was not performed.
Comparative example 2
The difference from example 2 is that no low-temperature plasma treatment was performed.
Comparative example 3
The difference from example 2 is that no high pressure treatment is carried out, i.e. the meat emulsion is directly subjected to subsequent enzymatic hydrolysis by the addition of enzymes after the addition of water.
Comparative example 4
The difference from example 2 is that neither low-temperature plasma treatment nor high-pressure treatment was performed.
Experimental example 1 measurement of salty taste of proteolysis product
1. Sensory evaluation
(1) Evaluation method
20 evaluators were randomly selected from the sensory evaluators library, and the appearance, smell, taste and smell of the proteolytic enzyme products of examples 1 to 3 and comparative examples 1 to 4 were evaluated by a simple descriptive test method, and the salty taste enhancement effect of the proteolytic enzyme products of examples 1 to 3 and comparative examples 1 to 4 on 50mmol/L NaCl solution was evaluated by a magnitude estimation method.
The evaluation method of appearance, smell, taste and fishy smell of the protease hydrolysate comprises the following steps:
s1, firstly, carrying out certain training to enable all sensory evaluation personnel to describe appearance, smell, taste and fishy smell of a sample by using the same concept;
s2, taking the proteolysis product of the comparative example 4 as a reference during sensory evaluation, writing the characteristics of each sample clearly by each sensory evaluator during the evaluation, and after all the evaluations are finished, discussing under the supervision of a group leader to obtain a comprehensive conclusion, thereby obtaining the results shown in the table 1.
The evaluation method of the salty taste enhancement effect of the proteolysis products on the 50mmol/L NaCl solution comprises the following steps:
s1, firstly, guiding sensory evaluation personnel to master the basic concept of a quantity value estimation method through evaluating the area of a geometric figure, then preparing sodium chloride solutions with 0, 10, 20, 30, 40, 50, 60, 70, 80, 90 and 100mmol/L series of gradients, and enabling the evaluation personnel to taste the sodium chloride solutions with the series of gradient concentrations so as to know the salty taste intensity of the sodium chloride solutions with different concentrations;
s2, adding water into the proteolysis products of the examples 1-3 and the comparative examples 1-4 respectively to ensure that the protein content of all samples is 20mg/mL, and adding NaCl to ensure that Na in the solution is added + The content was adjusted to 50mmol/L, and the specific saline taste intensity value (mmol/L NaCl) of the sample to be tested was given after tasting one by the assessors, and the results recorded by all the assessors were averaged to obtain the results shown in Table 1.
It should be noted that, 1h before the sensory evaluation experiment, sensory evaluation personnel can not eat other foods and beverages with pungent taste and can not be too tired; the sample to be tested is numbered by adopting three random combination numbers; in addition, in the evaluation experiment of the salty taste enhancement effect, an evaluator only evaluates the salty taste intensity of the sample solution without considering other sensory characteristics.
(2) Evaluation results
TABLE 1 sensory evaluation results of the proteolytic digests
As can be seen from Table 1, the protein zymolytes of the examples 1 to 3 are remarkably superior to the comparative examples 1 to 4 in appearance, smell, taste, fishy smell and other aspects; and the proteolysis products of the embodiments 1 to 3 with the protein content of 20mg/mL can improve the salty taste intensity of 50mmol/L NaCl solution to 72.8 to 78.6mmol/L, which is increased by 45.6 to 57.2 percent compared with 50mmol/L NaCl solution. Therefore, the protein zymolyte obtained by the method can be used as a high-quality salty taste enhancer through high-pressure synergistic low-temperature plasma treatment, and the salty taste of the seasoning is remarkably improved.
2. Electronic tongue analysis
(1) Analytical method
Taste signals of 50mmol/L NaCl solutions (in which the protein contents were all 20 mg/mL) to which the proteolytic digests of examples 1 to 3 and comparative examples 1 to 4 were added were examined using a TS-5000Z taste analysis system of the Japanese Insent company using a 50mmol/L sodium chloride solution as a reference solution. Four taste sensors of the TS-5000Z taste analysis system interact with the taste substances respectively to cause the change of the potential of the artificial lipid membrane, and the potential is taken as an output signal of the sensor to be transmitted to a computer for analysis, so that the delicate flavor, the salty flavor, the bitter flavor, the astringent flavor and the thick flavor of the sample are evaluated. The four taste sensors are: the taste sensor comprises an umami sensor AAE for reflecting the umami taste and the thick taste of a sample, a salty taste sensor CT0 for reflecting the salty taste of the sample, a bitter taste sensor C00 for reflecting the bitter taste of the sample and an astringent taste sensor AE1 for reflecting the astringent taste of the sample. Both taste sensor and reference electrode need to be activated before use to ensure stability of the data.
(2) Analysis results
The detection result is shown in fig. 1, and it can be seen that, compared with 50mmol/L sodium chloride solution (reference solution) without adding proteolysis, 50mmol/L sodium chloride solution with adding proteolysis of examples 1 to 3 and comparative examples 1 to 4 has improved salty taste and delicate taste, wherein the salty taste improvement effect of proteolysis of examples 1 to 3 is significantly better than that of comparative examples 1 to 4; and the proteolysis products of examples 1 to 3 also significantly reduced astringency compared to comparative examples 2 and 3.
3. Peptide molecular weight distribution determination
(1) Measurement method
The molecular weight distribution of the peptides of the proteolysis products of the examples 1 to 3 and the comparative examples 1 to 4 is determined by adopting a high performance liquid chromatography, and the specific conditions are as follows: TSKG2000SWXL (7.8X 300mm,5 μm) gel chromatography column with mobile phase 80:20 of a 0.125% (v/v) trifluoroacetic acid solution and acetonitrile at a flow rate of 1.0mL/min and a detection wavelength of 220nm. The peptide standard products are cytochrome C (12384 Da), aprotinin (6511 Da), somatostatin (1637 Da), asp-Phe-Pro-Ala-Leu (561 Da) and Leu-Val-Phe (377 Da). The fitting linear equation of the molecular weight logarithm value and the retention time of the peptide standard product is as follows: y = -2.587x +17.002 (R) 2 = 0.99), wherein, y is the logarithm of the molecular weight of the peptide standard; x-retention time.
(2) Measurement results
As shown in fig. 2, it can be seen that the release amounts of the peptides of 500-1000 Da and <500Da in the proteolytic digests of examples 1-3 are significantly greater than those in comparative examples 1-4, which indicates that the treatment method of the present invention can significantly promote the release of the peptides of 1000Da and <500Da in the proteolytic digests, and the method can significantly promote the salty taste enhancement effect of the proteolytic digests probably related to the promotion of the release of the peptides of <1000Da because the peptides of <1000Da are mainly related to the salty taste enhancement effect or salty taste enhancement effect.
In conclusion, the obtained proteolysis product can obviously improve the salty taste (the salty taste intensity of 50mmol/L NaCl solution can be increased by 45.6-57.2% by the proteolysis product with the protein content of 20 mg/mL) and the delicate flavor of the seasoning through the high-pressure synergistic low-temperature plasma treatment, can be used as a high-quality salty taste enhancer and delicate flavor enhancer, and is beneficial to guaranteeing the diet health of consumers.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A preparation method of a proteolysis product is characterized in that animals and plants are crushed, water is added, and then high-pressure treatment, enzymolysis and low-temperature plasma treatment are sequentially carried out to obtain the proteolysis product;
wherein the pressure of the high-pressure treatment is 100-400 MPa.
2. The method according to claim 1, wherein the low-temperature plasma treatment is: low-temperature plasma treatment is carried out for 5-15 min at the power of 30-40 w and the temperature of-2 ℃.
3. The method according to claim 1, wherein the gas generating the low-temperature plasma treatment is air.
4. The method according to claim 3, wherein the flow rate of the gas is 3 to 5L/min.
5. The method according to claim 1, wherein the high pressure treatment is carried out for 15 to 25min.
6. The method according to claim 1, wherein the enzyme used for the enzymatic hydrolysis is a protease.
7. The method of claim 6, wherein the enzyme comprises one or more of papain, trypsin, or flavourzyme.
8. The preparation method according to claim 1, wherein the enzymatic hydrolysis is: enzymolysis is carried out for 3.5 to 4.5 hours at the temperature of between 45 and 55 ℃.
9. A proteolytic digest prepared by the process of any one of claims 1 to 8.
10. Use of the proteolytic digest of claim 9 as and/or in the preparation of a salty taste enhancer.
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| CN117297041A (en) * | 2023-09-28 | 2023-12-29 | 杭州康源食品科技有限公司 | Industrial preparation process and application of hydrolyzed yolk powder capable of promoting growth of tibia |
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| CN117297041A (en) * | 2023-09-28 | 2023-12-29 | 杭州康源食品科技有限公司 | Industrial preparation process and application of hydrolyzed yolk powder capable of promoting growth of tibia |
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