CN112956668A - Method for preparing wall-broken bee pollen by using ultrasonic-assisted biological enzyme method - Google Patents
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- 238000000034 method Methods 0.000 title claims abstract description 36
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- 102000004190 Enzymes Human genes 0.000 title claims abstract description 30
- 229940088598 enzyme Drugs 0.000 claims abstract description 29
- 108010059892 Cellulase Proteins 0.000 claims abstract description 17
- 108090000526 Papain Proteins 0.000 claims abstract description 17
- 108010059820 Polygalacturonase Proteins 0.000 claims abstract description 17
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims description 2
- 230000008014 freezing Effects 0.000 claims description 2
- 239000008187 granular material Substances 0.000 claims description 2
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 2
- 238000000527 sonication Methods 0.000 claims 1
- 235000015097 nutrients Nutrition 0.000 abstract description 17
- 230000000694 effects Effects 0.000 description 21
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L21/00—Marmalades, jams, jellies or the like; Products from apiculture; Preparation or treatment thereof
- A23L21/20—Products from apiculture, e.g. royal jelly or pollen; Substitutes therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/06—Enzymes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/30—Physical treatment, e.g. electrical or magnetic means, wave energy or irradiation
- A23L5/32—Physical treatment, e.g. electrical or magnetic means, wave energy or irradiation using phonon wave energy, e.g. sound or ultrasonic waves
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention relates to a method for preparing wall-broken bee pollen by an ultrasonic-assisted biological enzyme method, which comprises the following steps: firstly, the bee pollen is subjected to ultrasonic treatment, and is subjected to stepwise enzymolysis by using pectinase, papain and cellulase, so that the comprehensive wall breaking treatment of the bee pollen is realized, and the purpose of improving the nutrient release rate of the bee pollen is achieved. According to the invention, the wall breaking treatment is carried out on the bee pollen by adopting an ultrasonic-assisted biological enzyme method, on one hand, the excessive damage to the bee pollen in the wall breaking treatment is improved by the biological enzyme method with mild conditions, the loss of nutrient components in the bee pollen is reduced, and the nutrient value and the product quality of the bee pollen are improved; on the other hand, on the premise of ensuring the nutrient components of the bee pollen, the ultrasonic method is used for breaking the wall of the bee pollen, so that the wall breaking efficiency of the bee pollen is improved, and the full release of the nutrient components is facilitated; the method is natural, safe and effective, and can further promote the comprehensive utilization of bee resources.
Description
(I) technical field
The invention relates to a method for preparing wall-broken bee pollen by an ultrasonic-assisted biological enzyme method.
(II) background of the invention
Bee pollen is irregular oblate spheroid particle formed by mixing pollen balls collected by bees with nectar, self glandular secretion and saliva. Bee pollen is known as "the only perfect food" because it contains rich nutrients such as protein, fatty acid, carbohydrate, amino acids, minerals, vitamins and polyphenols. China is the biggest bee product production and export country in the world, the bee pollen yield is high, and the export total amount is in an increasing state every year. In addition, China has wide territory, rich natural conditions, various bee source plants, wide distribution range and full-year blossoming, each bee colony can produce 3-30 kg of pollen every year, about 700 ten thousand bee colonies in China can produce about 21 ten thousand tons of bee pollen every year, and the bee pollen is a resource with rich sources. With the improvement of living standard of people and the enhancement of health care consciousness of people, the demand of bee pollen is increased day by day, so the research on the nutrient components and the application of the bee pollen can accelerate the high-efficiency utilization and the comprehensive development of the bee pollen.
The bee pollen contains rich nutrient substances, but the existence of the bee pollen wall not only prevents the human body from digesting and absorbing the bee pollen active ingredients, but also prevents the high-value utilization of the bee pollen. The wall breaking of bee pollen means that the structure of the bee pollen is damaged to a certain degree to release the nutrient substances contained in the bee pollen. Bee pollen wall breaking generally takes three forms: the pollen wall is broken from the germination hole; pollen wall breaks elsewhere; the pollen wall is completely crushed. It is generally believed that the nutrients of bee pollen can be fully released by completely pulverizing the structure of bee pollen, thereby exerting the maximum nutritional value.
At present, the wall breaking method of bee pollen mainly comprises a physical method, a chemical method, a biological method and a composite method. At present, the existing single wall breaking technology has certain limitations, and has the defects of complex process, high production cost, low wall breaking effect, serious damage to nutrient components and the like. If the ultrasonic time of the ultrasonic wall-breaking method is too long, the water-soluble nutrient contents in the bee pollen can be destroyed; the bee pollen wall breaking rate treated by the biological enzymolysis method technology is low, and the fermentation time is long. Therefore, the invention aims at the problems of excessive damage and insufficient damage of the current bee pollen single wall breaking technology, and aims to adopt the biological enzymolysis method and the ultrasonic wall breaking method, thereby overcoming the negative effect of a certain single process, achieving good wall breaking effect and effectively protecting the nutrient components in the bee pollen.
Disclosure of the invention
Based on the problems, the invention provides a method for preparing wall-broken bee pollen by using an ultrasonic-assisted biological enzyme method, which realizes the comprehensive wall-breaking treatment of the bee pollen with quick and efficient wall breaking of the bee pollen and less loss of nutrient components of the bee pollen.
The technical scheme adopted by the invention is as follows:
a method for preparing wall-broken bee pollen by using an ultrasonic-assisted biological enzyme method comprises the following steps:
(1) mechanically breaking cell wall of bee pollen granule to obtain bee pollen powder;
(2) mixing the bee pollen powder obtained in the step (1) with distilled water according to the feed liquid mass ratio of 1: 10-1: 30, and adjusting the pH value to 6.0 +/-0.5 to obtain a suspension; adjusting the pH value by using sodium hydroxide or hydrochloric acid;
(3) carrying out ultrasonic treatment on the suspension obtained in the step (2) at the temperature of 30-50 ℃ for 5-30 min to obtain ultrasonic treatment suspension; the ultrasonic power is 600-800W, and the ultrasonic frequency is 50-80 kHz;
(4) adding pectinase, papain and cellulase into the ultrasonic suspension obtained in the step (3) step by step, shaking uniformly, and performing enzymolysis for 10-20 hours at the temperature of 40-70 ℃ and the pH value of 3-8 to obtain an zymolyte; the addition amount of each enzyme is 1-10% of the mass of the pollen;
(5) pre-freezing the zymolyte obtained in the step (5) at the temperature of-80 to-70 ℃ for 18 to 20 hours, and then carrying out vacuum freeze drying at the temperature of-60 to-40 ℃ and under the pressure of 180 to 220mTorr for 70 to 75 hours to obtain the wall-broken bee pollen.
Preferably, the ultrasonic treatment conditions in step (3) are as follows: the ultrasonic power is 800W, the ultrasonic frequency is 80kHz, the ultrasonic temperature is 30 ℃, and the ultrasonic time is 15 min.
Preferably, in the step (4), the addition amount of the pectinase is 5%, the addition amount of the papain is 10%, and the addition amount of the cellulase is 8%.
More preferably, in the step (4), pectinase is added firstly, the enzymolysis temperature is 45 ℃, the enzymolysis pH is 5.0, and the enzymolysis time is 6 hours; then adding papain, wherein the enzymolysis temperature is 55 ℃, the enzymolysis pH is 5.0, and the enzymolysis time is 6 h; and finally adding cellulase, wherein the enzymolysis temperature is 50 ℃, the enzymolysis pH is 6.0, and the enzymolysis time is 6 h.
The invention has the following beneficial effects: according to the invention, the wall breaking treatment is carried out on the bee pollen by adopting an ultrasonic-assisted biological enzyme method, on one hand, the excessive damage to the bee pollen in the wall breaking treatment is improved by the biological enzyme method with mild conditions, the loss of nutrient components in the bee pollen is reduced, and the nutrient value and the product quality of the bee pollen are improved; on the other hand, on the premise of ensuring the nutrient components of the bee pollen, the ultrasonic method is used for breaking the wall of the bee pollen, so that the wall breaking efficiency of the bee pollen is improved, and the full release of the nutrient components is facilitated; the method is natural, safe and effective, and can further promote the comprehensive utilization of bee resources.
(IV) description of the drawings
FIG. 1 is an electron microscope image of cell wall broken form of bee pollen, wherein a is an electron microscope image of rape bee pollen before cell wall breaking, and b is an electron microscope image of rape bee pollen after cell wall breaking.
(V) detailed description of the preferred embodiments
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto:
in the following examples, the wall-breaking rate of bee pollen is calculated by microscopic examination, i.e.,% wall-breaking rate is wall-breaking pollen count/total pollen count in visual field × 100%
In the following examples, cellulase (activity 10000U/g) was purchased from Shanghai Michelin, Inc., papain (activity 10000U/g) was purchased from Shanghai Shuyang Biotechnology, Inc., and pectinase (activity 10000U/g) was purchased from national drug group chemical reagent, Inc.
Examples 1 to 5: influence of different ultrasonic time on wall breaking effect of bee pollen
5g of bee pollen after mechanical wall breaking is taken in a 250mL beaker, distilled water is added according to the material-liquid ratio of 1:20, and the mixture is evenly mixed and then is subjected to ultrasonic treatment. The ultrasonic temperature is 40 ℃, the ultrasonic power is 800W, the ultrasonic frequency is 80kHz, the ultrasonic time is 5min, 10 min, 15min, 20 min and 25min respectively, and the bee pollen wall breaking rate is calculated respectively, and the results are listed in Table 1.
Table 1: influence of different ultrasonic time on bee pollen wall breaking rate
Examples 6 to 8: influence of different ultrasonic temperatures on wall breaking effect of bee pollen
5g of wall-broken bee pollen is taken and put into a 250mL beaker, distilled water is added according to the material-liquid ratio of 1:20, and the mixture is evenly mixed and then is subjected to ultrasonic treatment. The ultrasonic time is 15min, the ultrasonic power is 800W, the ultrasonic frequency is 80kHz, the ultrasonic temperature is 30 ℃, 40 ℃ and 50 ℃, the bee pollen wall breaking rate is respectively calculated, and the results are listed in Table 2.
Table 2: influence of different ultrasonic time on bee pollen wall breaking rate
Examples 9 to 11: influence of different feed liquid ratios on wall breaking effect of bee pollen
Taking 5g of wall-broken bee pollen in a 250mL beaker, wherein the material-liquid ratio is 1:10, 1:15, 1:20, 1:25 and 1:30 respectively. The ultrasonic power is 800W, the ultrasonic frequency is 80kHz, the ultrasonic time is 15min, and the ultrasonic temperature is 30 ℃. Respectively comparing the influence of different feed liquid ratios on the wall breaking effect of bee pollen. The conditions for the enzymatic hydrolysis of the three enzymes are as follows: the addition amount of the pectinase is 5 percent, the enzymolysis temperature is 45 ℃, the enzymolysis pH is 5.0, and the enzymolysis time is 4 hours; the additive amount of the papain is 5 percent, the enzymolysis temperature is 57 ℃, the enzymolysis time is 4 hours, and the enzymolysis pH is 6.0; the addition amount of cellulase is 5%, the enzymolysis temperature is 50 ℃, the enzymolysis time is 4h, and the enzymolysis pH is 5.0; the wall breaking rate of bee pollen was calculated separately and the results are shown in Table 3.
Table 3: influence of different feed liquid ratios on bee pollen wall breaking rate
Examples 12 to 14: influence of different enzyme enzymolysis pH values on wall breaking effect of bee pollen
Taking 5g of wall-broken bee pollen in a 250mL beaker, the material-liquid ratio is 1:20, the ultrasonic power is 800W, the ultrasonic frequency is 80kHz, the ultrasonic time is 15min, and the ultrasonic temperature is 30 ℃. Respectively comparing the influence of different enzyme enzymolysis pH values on the bee pollen wall breaking effect. The addition amount of the pectinase is 5 percent, the enzymolysis temperature is 45 ℃, the enzymolysis time is 4 hours, and the pH values of the pectinase and the pectinase are respectively 3.0, 4.0, 5.0, 6.0 and 7.0; the additive amount of the papain is 5 percent, the enzymolysis temperature is 57 ℃, the enzymolysis time is 4 hours, and the papain is respectively subjected to enzymolysis at different pH values of 3.0, 4.0, 5.0, 6.0 and 7.0; the addition amount of the cellulase is 5 percent, the enzymolysis temperature is 50 ℃, the enzymolysis time is 4 hours, and the pH values of the cellulase and the cellulase are respectively 4.0, 5.0, 6.0, 7.0 and 8.0 in different enzymolysis processes; the wall breaking rate of bee pollen was calculated separately and the results are shown in Table 4.
Table 4: influence of different enzyme enzymolysis pH values on bee pollen wall breaking rate
Examples 15 to 17: influence of different enzyme enzymolysis time on wall breaking effect of bee pollen
Taking 5g of wall-broken bee pollen in a 250mL beaker, the material-liquid ratio is 1:20, the ultrasonic power is 800W, the ultrasonic frequency is 80kHz, the ultrasonic time is 15min, and the ultrasonic temperature is 30 ℃. Respectively comparing the influence of different enzyme enzymolysis time on the wall breaking effect of bee pollen. The addition amount of the pectinase is 5%, the enzymolysis temperature is 45 ℃, the enzymolysis pH is 5, and the enzymolysis time is 2.0h, 4.0h, 6.0h, 8.0h and 10.0h respectively; the additive amount of the papain is 5 percent, the enzymolysis temperature is 57 ℃, the enzymolysis pH is 6, and the papain is subjected to enzymolysis for 2.0h, 4.0h, 6.0h, 8.0h and 10.0h respectively; the addition amount of the cellulase is 5%, the enzymolysis temperature is 50 ℃, the enzymolysis pH is 5, and the enzymolysis time is 2.0h, 4.0h, 6.0h, 8.0h and 10.0h respectively; the wall breaking rate of bee pollen was calculated separately and the results are shown in Table 5.
Table 5: influence of different enzyme enzymolysis time on bee pollen wall breaking rate
Examples 18 to 20: influence of different enzyme enzymolysis temperatures on wall breaking effect of bee pollen
Taking 5g of wall-broken bee pollen in a 250mL beaker, the material-liquid ratio is 1:20, the ultrasonic power is 800W, the ultrasonic frequency is 80kHz, the ultrasonic time is 15min, and the ultrasonic temperature is 30 ℃. Respectively comparing the influence of different enzyme enzymolysis temperatures on the wall breaking effect of bee pollen. 5 percent of pectinase, 6 hours of enzymolysis, 5.0 of enzymolysis pH, 25 ℃, 35 ℃, 45 ℃, 55 ℃ and 65 ℃; the addition amount of papain is 5%, the enzymolysis time is 6h, the enzymolysis pH is 6.0, and the enzymolysis temperature is 35 deg.C, 45 deg.C, 55 deg.C, 65 deg.C, and 75 deg.C; the addition amount of cellulase is 5%, the enzymolysis time is 6h, the enzymolysis pH is 5.0, and the enzymolysis temperature is 30 ℃, 40 ℃, 50 ℃, 60 ℃ and 70 ℃; the wall breaking rate of bee pollen was calculated separately and the results are shown in Table 6.
Table 6: influence of different enzyme enzymolysis temperatures on bee pollen wall breaking rate
Examples 21 to 23: influence of different enzyme addition amounts on bee pollen wall breaking effect
Taking 5g of wall-broken bee pollen in a 250mL beaker, the material-liquid ratio is 1:20, the ultrasonic power is 800W, the ultrasonic frequency is 80kHz, the ultrasonic time is 15min, and the ultrasonic temperature is 30 ℃. Respectively comparing the influence of different enzyme addition amounts on the bee pollen wall breaking effect. The addition amount of pectinase is 1%, 5%, 8%, 10% and 15%, the enzymolysis temperature is 45 ℃, the enzymolysis time is 6h, and the enzymolysis pH is 5.0; the additive amount of the papain is 1%, 5%, 8%, 10% and 15%, the enzymolysis temperature is 57 ℃, the enzymolysis time is 6h, and the enzymolysis pH is 6.0; the addition amount of cellulase is 1%, 5%, 8%, 10% and 15%, the enzymolysis temperature is 50 ℃, the enzymolysis time is 4h, and the enzymolysis pH is 6.0; the wall breaking rate of bee pollen was calculated separately and the results are shown in Table 7.
Table 7: influence of different enzyme addition amounts on bee pollen wall breaking rate
Examples 24 to 25: influence of enzyme addition mode on wall breaking effect of bee pollen
Taking 5g of wall-broken bee pollen in a 250mL beaker, the material-liquid ratio is 1:20, the ultrasonic power is 800W, the ultrasonic frequency is 80kHz, the ultrasonic time is 15min, and the ultrasonic temperature is 30 ℃. Respectively comparing the influence of different enzyme addition amounts on the bee pollen wall breaking effect. The three enzymes were added simultaneously in the following manner: adding 5% of pectinase, papain and cellulase respectively, performing enzymolysis at 45 deg.C for 6h and pH of 5, and calculating cell wall breaking rate of bee pollen after enzymolysis; the stepwise addition was as follows: the addition amount of the pectinase is 5 percent, the enzymolysis temperature is 45 ℃, the enzymolysis time is 6 hours, and the enzymolysis pH is 5.0; adding 10% of papain, carrying out enzymolysis at 55 deg.C for 6h, and carrying out enzymolysis at pH5.0 respectively; the addition amount of cellulase is 8 percent, the enzymolysis temperature is 50 ℃, the enzymolysis time is 6 hours, and the enzymolysis pH is 6.0; the wall breaking rate of bee pollen was calculated separately and the results are shown in Table 8.
Table 8: influence of enzyme addition mode on wall breaking effect of bee pollen
Example 26: analysis of main effective components before and after wall breaking treatment of bee pollen
The analysis of each effective component before and after wall breaking is shown in table 9. Before wall breaking, the total phenol content in bee pollen is 4.41 + -0.23 g/100g, the total flavone content is 28.57 + -2.33 g/100g, the crude protein content is 18.5 + -2.3 g/100g, the total sugar (calculated by reducing sugar) content is 6.7 + -1.2 g/100g, the free amino acid content is 0.735 + -0.02 g/100g, after wall breaking by ultrasonic-assisted biological enzyme method, the total phenol content is 7.26 + -0.33 g/100g, the total flavone content is 49.48 + -4.37 g/100g, the protein content is 26.3 + -3.1 g/100g, the crude fat content is 34.8 + -2.6 g/100g, the total sugar (calculated by reducing sugar) is 47.8 + -3.6 g/100g, and the free amino acid content is increased to 0.89 + -0.05 g/100 g.
Table 9: analysis of main effective components of bee pollen before and after wall breaking
Claims (4)
1. A method for preparing wall-broken bee pollen by using an ultrasonic-assisted biological enzyme method comprises the following steps:
(1) mechanically breaking cell wall of bee pollen granule to obtain bee pollen powder;
(2) mixing the bee pollen powder obtained in the step (1) with distilled water according to the feed liquid mass ratio of 1: 10-1: 30, and adjusting the pH value to 6.0 +/-0.5 to obtain a suspension;
(3) carrying out ultrasonic treatment on the suspension obtained in the step (2) at the temperature of 30-50 ℃ for 5-30 min to obtain ultrasonic treatment suspension; the ultrasonic power is 600-800W, and the ultrasonic frequency is 50-80 kHz;
(4) adding pectinase, papain and cellulase into the ultrasonic suspension obtained in the step (3) step by step, shaking uniformly, and performing enzymolysis for 10-20 hours at the temperature of 40-70 ℃ and the pH value of 3-8 to obtain an zymolyte;
(5) pre-freezing the zymolyte obtained in the step (5) at the temperature of-80 to-70 ℃ for 18 to 20 hours, and then carrying out vacuum freeze drying at the temperature of-60 to-40 ℃ and under the pressure of 180 to 220mTorr for 70 to 75 hours to obtain the wall-broken bee pollen.
2. The method of claim 1, wherein the sonication conditions of step (3) are: the ultrasonic power is 800W, the ultrasonic frequency is 80kHz, the ultrasonic temperature is 30 ℃, and the ultrasonic time is 15 min.
3. The method according to claim 1, wherein in the step (4), the addition amount of the pectinase is 5%, the addition amount of the papain is 10%, and the addition amount of the cellulase is 8%.
4. The method according to claim 3, wherein in the step (4), pectinase is added firstly, the enzymolysis temperature is 45 ℃, the enzymolysis pH is 5.0, and the enzymolysis time is 6 h; then adding papain, wherein the enzymolysis temperature is 55 ℃, the enzymolysis pH is 5.0, and the enzymolysis time is 6 h; and finally adding cellulase, wherein the enzymolysis temperature is 50 ℃, the enzymolysis pH is 6.0, and the enzymolysis time is 6 h.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113633604A (en) * | 2021-08-30 | 2021-11-12 | 合肥赛为智慧医疗有限公司 | Mouthwash for preventing and treating gingivitis and preparation method thereof |
| CN113632976A (en) * | 2021-08-23 | 2021-11-12 | 浙江亚林生物科技股份有限公司 | Preparation method of pollen pini with wall removed |
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| CN113633604A (en) * | 2021-08-30 | 2021-11-12 | 合肥赛为智慧医疗有限公司 | Mouthwash for preventing and treating gingivitis and preparation method thereof |
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