Bee pollen biological wall breaking method with high wall breaking rate

Abstract

The invention belongs to the field of pollen wall breaking, and particularly relates to a bee pollen biological wall breaking method with a high wall breaking rate. On the basis of an enzymolysis and ultrasonic combined bee pollen wall breaking process, the bee pollen is subjected to cold and hot treatment before enzymolysis, and it is found that the wall breaking rate of the bee pollen can be remarkably increased. Furthermore, a protective agent composed of one or more of glutamic acid, glucose, mannitol and trehalose is added during heating treatment, and it is found that nutrient substances in the bee pollen can be prevented from being damaged during cold and hot treatment.

Description

Technical Field

[0001] The invention belongs to the field of pollen wall breaking, and particularly relates to a bee pollen biological wall breaking method with a high wall breaking rate. Background Art

[0002] Bee pollen refers to the pollen mass brought back by bees when collecting honey, and the pollen formed after storage and fermentation in the honeycomb, but the nutrients are mainly distributed inside the bee pollen wall, which is wrapped by the hard outer wall and inner wall that are acid-resistant, alkali-resistant and corrosion-resistant. The outer wall of bee pollen is mainly composed of sporopollenin and cellulose, and the inner wall mainly contains cellulose and pectin. The existence of the bee pollen wall hinders the absorption and utilization of the internal nutrients of bee pollen by the human body, so it is necessary to carry out wall-breaking treatment on bee pollen. The currently commonly used bee pollen wall-breaking methods are mainly fermentation wall-breaking method, temperature difference wall-breaking method, mechanical wall-breaking method and supercritical carbon dioxide, etc., but these wall-breaking methods each have some defects, such as many nutrients are lost, the wall-breaking rate is not ideal, the wall-breaking time is long, the wall-breaking efficiency is not high, and it is easy to pollute. This area still needs a bee pollen wall-breaking method that avoids damaging nutrients while achieving a high wall-breaking rate. Summary of the invention

[0003] In order to solve the above problems, the present invention provides a bee pollen biological wall breaking method with a high wall breaking rate, which comprises the following steps:

[0004] (1) freezing bee pollen at -20 to -30°C for 5 to 8 days, stirring it in an aqueous solution containing a protective agent having a pH value of 7.5 to 8.5 at 65 to 75°C for 20 to 30 minutes, and then cooling it to room temperature to obtain a pretreated suspension;

[0005] (2) mixing the pretreated suspension obtained in step (1) with the composite enzyme, and performing enzymatic hydrolysis under ultrasound for 2 to 3 hours to obtain a cell wall-breaking liquid;

[0006] (3) vacuum freeze-drying the broken bee pollen liquid obtained in step (2) to obtain a finished broken bee pollen product.

[0007] Furthermore, the protective agent comprises one or more of glutamic acid, glucose, mannitol and trehalose.

[0008] Furthermore, the protective agent comprises glutamic acid, glucose, mannitol and trehalose.

[0009] Furthermore, the mass ratio of glutamate, glucose, mannitol and trehalose is 1:1:1:1.

[0010] Furthermore, the mass concentration of the protective agent in the aqueous solution is 10-20%.

[0011] Furthermore, in step (1), the solid-liquid ratio of bee pollen to aqueous solution is 1 g:5-10 ml.

[0012] Furthermore, in step (2), the mass ratio of bee pollen to protease in the pretreated suspension is 8 to 12:1; and the enzyme activity of the protease is 30,000 to 300,000 U.

[0013] Furthermore, the protease includes one or more of cellulase, pectinase and neutral protease.

[0014] Furthermore, the enzymatic hydrolysis is carried out at 45-55°C.

[0015] Furthermore, the power of the ultrasound is 800-1200w, the frequency is 70-90kHz, the ultrasound working time is 0.5h, and the interval time is 0.5h.

[0016] The present invention also provides wall-broken bee pollen prepared by the method described herein.

[0017] Beneficial effects of the present invention

[0018] Enzymatic hydrolysis + ultrasound is a commonly used method to achieve a high pollen wall breaking rate, but its wall breaking rate still cannot reach more than 90%. For this reason, the present invention freezes and heats the bee pollen before enzymatic hydrolysis, which can significantly improve the wall breaking rate of the bee pollen. However, the temperature difference wall breaking method easily destroys the nutrients in the bee pollen. For this reason, the present invention innovatively adds a protective agent during the temperature treatment, and finds that the protective agent composed of glutamic acid, glucose, mannitol and trehalose can prevent the nutrients in the bee pollen from being destroyed during cold and hot treatments. DETAILED DESCRIPTION

[0019] The present invention is further described below with reference to specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0020] Example 1

[0021] A bee pollen biological wall breaking method with a high wall breaking rate comprises the following steps:

[0022] (1) Rapeseed bee pollen (purchased from the Bee Research Institute of the Chinese Academy of Agricultural Sciences) was first soaked in 10vt% alcohol for 60s and then soaked in 75vt% alcohol for 80s for disinfection. After natural drying, it was frozen at -30°C for 5 days, added to an aqueous solution containing 15wt% of a protective agent (composed of glutamic acid, glucose, mannitol and trehalose in a mass ratio of 1:1:1:1) at a solid-liquid ratio of 1g:8mL at pH 8.0, stirred at 70°C for 30 minutes, and then cooled to room temperature to obtain a pretreated suspension;

[0023] (2) mixing the pretreated suspension obtained in step (1) with a composite enzyme (60,000 U of composite enzyme, composed of cellulase, pectinase and neutral protease with an enzyme activity value of 1:1:1), and performing enzymatic hydrolysis at 50° C. for 2 h, with an ultrasonic power of 1000 W, an ultrasonic frequency of 80 kHz, an ultrasonic working time of 0.5 h, and an interval of 0.5 h to obtain a cell wall-breaking liquid;

[0024] (3) The wall-breaking liquid obtained in step (2) is vacuum freeze-dried for 24 hours, and then ground into powder to obtain a finished wall-breaking bee pollen product.

[0025] Example 2

[0026] A bee pollen biological wall breaking method with a high wall breaking rate comprises the following steps:

[0027] (1) Rapeseed bee pollen (purchased from the Bee Research Institute of the Chinese Academy of Agricultural Sciences) was first soaked in 10vt% alcohol for 60s and then soaked in 75vt% alcohol for 80s for disinfection. After natural drying, it was frozen at -20°C for 8 days, added to an aqueous solution containing 20wt% of a protective agent (composed of glutamic acid, glucose, mannitol and trehalose in a mass ratio of 1:1:1:1) at a solid-liquid ratio of 1g:5ml at pH 8.0, stirred at 70°C for 30 minutes, and then cooled to room temperature to obtain a pretreated suspension;

[0028] (2) mixing the pretreated suspension obtained in step (1) with a protease (90,000 U of a composite enzyme consisting of cellulase, pectinase and neutral protease with an enzyme activity value of 1:1:1), and performing enzymatic hydrolysis at 50° C. for 3 h under ultrasound, with an ultrasound power of 1000 W, an ultrasound frequency of 80 kHz, an ultrasound working time of 0.5 h, and an interval of 0.5 h to obtain a cell wall-breaking liquid;

[0029] (3) The wall-breaking liquid obtained in step (2) is vacuum freeze-dried for 24 hours, and then ground into powder to obtain a finished wall-breaking bee pollen product.

[0030] Comparative Examples 1-11: Screening of protective agent ingredients

[0031] The rape bee pollen was subjected to cold and hot treatment according to the method of step (1) of Example 1, wherein no protective agent was used in Comparative Example 1, and the protective agents used in Comparative Examples 2-11 were sucrose, glutamic acid, lactose, mannitol, glycine, glucose, sorbitol, glycerol, trehalose, and dimethyl sulfoxide with a mass concentration of 15%, respectively. The obtained pre-treated suspension was vacuum freeze-dried, and the β-carotene content was determined according to the high performance liquid chromatography method in accordance with GB / T 5009.83-2003 (Determination of carotene in food). The results are shown in Table 1.

[0032] Table 1

[0033]

[0034] From the results in Table 1, it can be seen that when glutamic acid, glucose, mannitol and trehalose are used as protective agents, the stability of β-carotene in bee pollen treated with cold and hot treatments is the best.

[0035] Comparative Examples 12-22

[0036] According to the method of step (1) of Example 1, rape bee pollen was subjected to cold and hot treatment, wherein the protective agents used in Comparative Examples 12-22 were glutamic acid and glucose, glutamic acid and mannitol, glutamic acid and trehalose, glucose and mannitol, glucose and trehalose, mannitol and trehalose, glutamic acid, glucose and mannitol, glutamic acid, glucose and trehalose, glucose, mannitol and trehalose, glutamic acid, mannitol and trehalose, and glutamic acid, glucose, mannitol and trehalose in equal mass ratios. After the obtained pre-treated suspension was vacuum freeze-dried, the β-carotene content was determined according to the high performance liquid chromatography method in GB / T 5009.83-2003 (Determination of carotene in food). The results are shown in Table 2.

[0037] Table 2

[0038]

[0039]

[0040] From the results in Table 2, it can be seen that when glutamic acid, glucose, mannitol and trehalose are used together as protective agents, the stability of β-carotene in bee pollen treated with cold and hot treatments is the best.

[0041] Comparative Example 23

[0042] A bee pollen biological wall breaking method, which differs from Example 1 in that no cold or hot pretreatment is performed in step (1) and no ultrasonic treatment is performed in step (2), and the remaining steps are the same as those in Example 1.

[0043] Comparative Example 24

[0044] A bee pollen biological wall breaking method, which differs from Example 1 in that no cold or hot pretreatment is performed in step (1), and the remaining steps are the same as those in Example 1.

[0045] Comparative Example 25

[0046] A bee pollen biological wall breaking method, which differs from Example 1 in that ultrasonic treatment is not performed in step (2), and the remaining steps are the same as those in Example 1.

[0047] The wall-breaking rates of the pollens prepared in Examples 1-2 and Comparative Examples 23-25 ​​were measured, and the results are shown in Table 3.

[0048] Table 3

[0049]

[0050] It can be seen from the results in Table 3 that the cell wall breaking process of cold and hot pretreatment + enzymatic hydrolysis + ultrasound adopted in the present invention can achieve a cell wall breaking rate of more than 90%.

[0051] It should be noted that the preferred embodiments of the present invention are given in the specification of the present invention, but the present invention can be implemented in many different forms and is not limited to the embodiments described in this specification. These embodiments are not intended to be additional limitations on the content of the present invention. The purpose of providing these embodiments is to make the understanding of the disclosure of the present invention more thorough and comprehensive. In addition, the above-mentioned technical features continue to be combined with each other to form various embodiments not listed above, which are all considered to be within the scope of the present invention specification; further, for ordinary technicians in this field, they can be improved or transformed according to the above description, and all these improvements and transformations should belong to the protection scope of the claims attached to the present invention.

Claims

1. A bee pollen biological wall breaking method with high wall breaking rate, characterized in that: The following steps are involved: (1) freezing bee pollen at -20 to -30°C for 5 to 8 days, stirring it in an aqueous solution containing a protective agent having a pH value of 7.5 to 8.5 at 65 to 75°C for 20 to 30 minutes, and then cooling it to room temperature to obtain a pretreated suspension; (2) mixing the pretreated suspension obtained in step (1) with the composite enzyme, and performing enzymatic hydrolysis under ultrasound for 2 to 3 hours to obtain a cell wall-breaking liquid; (3) vacuum freeze-drying the broken bee pollen liquid obtained in step (2) to obtain a finished broken bee pollen product.

2. The method according to claim 1, characterized in that The protective agent comprises one or more of glutamic acid, glucose, mannitol and trehalose.

3. The method according to claim 2, characterized in that The protective agent comprises glutamic acid, glucose, mannitol and trehalose; Furthermore, the mass ratio of glutamate, glucose, mannitol and trehalose is 1:1:1:

1.

4. The method according to claim 1, characterized in that: The mass concentration of the protective agent in the aqueous solution is 10-20%.

5. The method according to claim 1, characterized in that: In step (1), the material-liquid ratio of bee pollen to aqueous solution is 1 g:5-10 ml.

6. The method according to claim 1, characterized in that In step (2), the mass ratio of bee pollen to protease in the pretreatment suspension is 8 to 12:1; and the enzyme activity of the protease is 30,000 to 300,000 U.

7. The method according to claim 1, characterized in that The protease includes one or more of cellulase, pectinase and neutral protease.

8. The method according to claim 1, characterized in that The enzymatic hydrolysis is carried out at 45-55°C.

9. The method according to claim 1, characterized in that: The power of the ultrasound is 800-1200w, the frequency is 70-90kHz, the ultrasound working time is 0.5h, and the interval time is 0.5h.

10. A wall-broken bee pollen prepared by the method according to any one of claims 1 to 9.

Images