CN103598654B - Pollen active probiotic drink and preparation method thereof - Google Patents

Abstract

<b/>The invention provides a bee pollen active probiotic beverage and a preparation method thereof. The bee pollen active probiotic beverage is made of bee pollen as a raw material, mixed with a certain proportion of maltose and/or galactose, mixed and fermented by two kinds of probiotics, Lactobacillus BL1 and Lactobacillus BL2, and then formulated. In view of the characteristics of bee pollen raw materials with strong acidity and not easy to be utilized by microorganisms, the present invention selects two strains with strong acid resistance and various excellent properties, and keeps the viable count of probiotics within 30 days of the product at 10 ¹¹ cfu by controlling the fermentation process /L or more, and can biotransform bee pollen flavonoid glycosides to obtain a large amount of flavonoid aglycone, and improve the bioavailability of bee pollen flavonoids. At the same time, it is rich in other bee pollen and probiotic fermentation products. It is rich in nutrition, rich in fragrance, sweet and sour, and has the functions of beautifying and nourishing the skin, enhancing immunity, lowering cholesterol, regulating intestinal flora and preventing constipation.

Description

A kind of bee pollen active probiotic beverage and preparation method thereof

technical field

The invention relates to a bee pollen active probiotic beverage with health care function and a preparation method thereof, belonging to the technical field of health food.

Background technique

With the improvement of people's health awareness, health food has attracted more and more attention. Edible bee pollen has a long history in our country. Bee pollen is rich in amino acids, organic acids, unsaturated fatty acids, nucleic acids, enzymes, mineral elements, vitamins and other nutrients necessary for the human body, as well as flavonoids, polysaccharides and other health ingredients. Nutritional health and medicinal value. Modern studies have shown that bee pollen has various effects such as anti-oxidation, anti-fatigue, anti-aging, improving immunity, and improving memory. At present, bee pollen products in the health care market are relatively single, mostly in the form of powder, and have poor palatability , the bioavailability of various active ingredients is low. Therefore, it is of great significance to research and develop high-tech bee pollen products.

Probiotics refer to live bacterial preparations and their metabolites that improve the host’s micro-ecological balance and play a beneficial role in improving the host’s health level and health status. Probiotics exist in every corner of the earth. Beneficial bacteria or fungi in animals mainly include : Lactic acid bacteria, bifidobacteria, yeast, etc. Lactic acid bacteria are important probiotics. Active lactic acid bacteria will secrete substances that are beneficial to human health when they grow, so they have many benefits to the human body: 1. It has the effects of promoting gastric juice secretion, increasing appetite, and strengthening digestion; probiotics can convert milk 2. Maintain the ecological balance of intestinal flora, form a biological barrier, and inhibit the invasion of harmful bacteria to the intestinal tract; 3. Can reduce the production of certain carcinogens, so there are Anti-cancer effect; 4. It can inhibit the reproduction of spoilage bacteria in the intestinal tract, and weaken the toxins produced by spoilage bacteria in the intestinal tract; promote intestinal peristalsis and bacterial growth to change osmotic pressure by producing a large amount of short-chain fatty acids to prevent constipation; 5 1. It has the effect of lowering cholesterol, especially suitable for people with high blood fat; 6. Improves the immune function of the human body. Lactic acid bacteria can produce some substances that enhance the immune function, which can improve human immunity and prevent diseases. Generally speaking, whether it is after surgery, or after recovery from acute or chronic diseases, a large number of antibiotics have been taken or injected in order to treat diseases or prevent infection, which has greatly changed the intestinal flora, and some beneficial intestinal bacteria have been eliminated. Inhibit or kill, causing flora imbalance. Taking a certain amount of probiotic lactic acid bacteria every day can maintain the balance of intestinal flora and regulate the intestinal flora to normal levels.

The current reports on bee pollen fermented products are as follows. Qiao Lin (2005) carried out the research and development of bee pollen solid-state fermentation nutritional enhancer, conducted single-bacteria and mixed-bacteria solid-state fermentation tests on pollen by yeast, lactic acid bacteria, and Bacillus natto, and finally determined the most suitable strain for solid-state fermentation of bee pollen It is Bacillus natto and Lactobacillus acidophilus, and the two bacteria are mixed at a ratio of 1:1; the optimum fermentation method is shallow aerobic fermentation, and the inoculation method is to accept Bacillus beanus first, and then insert Lactobacillus acidophilus on the 3rd day of fermentation ; The inoculation amount was 10%, the water content of pollen was 35-40%, the fermentation temperature was 30°C, and the fermentation period was 8 days. At the same time, the scale-up test was carried out. The fermented pollen smells delicious, sweet and sour. The number of live bacteria of Bacillus natto in the pollen is 4.5×10 ⁷ cfu/g, and the number of live bacteria of Lactobacillus acidophilus is 2.7×10 ⁷ cfu/g. The crude protein increased by 5.9%, the crude fat decreased by 1.57%, the free amino acid increased by 0.02%, the total organic acid increased by 0.79g/100g, the pH value dropped to 4.2, and the maximum increase in lactic acid reached 0.20g/100g. Yang Wenchao et al. (2012) optimized the process of fermenting rapeseed bee pollen with lactic acid bacteria derived from bee grain, using solid-state fermentation, 13% of lactic acid bacteria inoculum, 10% of Monascus fermentation broth, 25% of water, and 33°C fermentation temperature. After fermentation, the palatability of bee pollen is improved, and the nutritional value is improved. Wang Cuihua et al. (2009) developed a functional yogurt containing bee pollen, adding 0.1% to 2% bee pollen to the yogurt, and the probiotics could be maintained at 10 ⁶ to 10 ⁹ cfu/ml within 21 days. Sun Shiyao (2008) and Chen Hunan (2011) also studied honey pollen wine respectively.

However, in the above studies, the solid-state fermentation of bee pollen generally has a large inoculum amount and a long fermentation time. The inoculum amount is more than 10%, and the fermentation time is about 1 week. Miscellaneous bacteria. Whether bee pollen is added to yogurt or used for wine making in liquid fermentation, the amount of bee pollen added is generally relatively small, and the amount of bee pollen generally does not exceed 2%. We have determined that the natural pH of bee pollen is between 4.5 and 5.0. At this lower pH value, microorganisms are easily inhibited by acidity and difficult to grow and ferment. It may also be the reason that the existing solid-state fermentation requires a relatively large inoculum size and a relatively long fermentation time, and the pH drop is not obvious at the same time. The amount of bee pollen should not be too large during liquid fermentation, otherwise it will inhibit the growth of microorganisms and fail to achieve the fermentation effect. Therefore, it is very important to choose a bacterial agent suitable for bee pollen fermentation.

Lactobacillus BL1 and BL2 are two strains of acid-resistant lactic acid bacteria (Liu Xiaoyong, Qiu Jiying, Sun Xin, etc.) isolated and identified from pickles by the Agricultural Products Research Institute of the Shandong Academy of Agricultural Sciences, the applicant for the patent of the present invention. The isolation of two strains of lactic acid bacteria in pickles Preliminary study on identification and mixed culture. Shandong Agricultural Science［J］, 2012, 44(9):85-89.). The above-mentioned article only reports on the isolation and identification of the above-mentioned two strains. However, the function and use of this strain have not been reported.

Contents of the invention

The invention overcomes the shortcomings in the fermentation process of the above-mentioned bee pollen products, and provides a bee pollen active probiotic beverage and a preparation method thereof. The bee pollen was cultured with two excellent strains of Lactobacillus BL1 and Lactobacillus BL2, and the main flavonoid glycosides were partially biotransformed into the corresponding aglycon-kaempferol, which improved the bioavailability of bee pollen flavonoids. At the same time, it contains a large number of other bee pollen functional ingredients and probiotic fermentation products, which have the functions of anti-oxidation, anti-fatigue, anti-aging, improving immunity, lowering cholesterol and regulating gastrointestinal balance.

One of the objects of the present invention is to provide a bee pollen active probiotic beverage with functions such as anti-oxidation, anti-fatigue, anti-aging, improving immunity, improving memory, and regulating gastrointestinal balance; the second object is to provide the bee pollen active probiotic beverage Process for the preparation of probiotic beverages.

One of the objectives of the present invention can be achieved through the following technical measures: a bee pollen active probiotic beverage is characterized in that it is made of the following raw materials in parts by weight: 3 to 10 bee pollen, 3 maltose and/or galactose ～5, water 100, cyclamate 0.030～0.065, acesulfame potassium 0.020～0.030; bee pollen, maltose and/or galactose are used as raw materials, and water is added to make a fermentation medium, which is passed through Lactobacillus BL1 and Lactobacillus BL2 Probiotics are fermented in equal volume ratio mixed in liquid state, post-ripened, and prepared by adding cyclamate and acesulfame potassium dissolved in water. The total amount of water used for two times before and after is 100 parts by weight, wherein the amount of water used to make the fermentation medium is >50 parts; the preferred weight ratio of the two is 99:1.

Two of the object of the present invention can be realized by following technical measures:

(1) Take each raw material according to the above parts by weight;

(2) Preparation of fermentation medium: remove impurities from bee pollen and crush it through a 40-80 mesh sieve, crush maltose and/or galactose through a 40 mesh sieve; mix the bee pollen, maltose and/or galactose evenly, and stir Slowly add 99 parts by weight of water, stir evenly and filter with gauze or 4 layers of gauze;

(3) Fermentation: Sterilize the fermentation culture at 115°C for 15-30 minutes, and cool it down to below 37°C; add 3-5% (preferably 4%) of the mixed strain expansion solution of the fermentation medium volume, 30-37°C Seal and culture for 18-30 hours; when the pH of the fermentation broth drops to 3.10-3.30, the total acid (calculated as lactic acid) reaches 5-7 g/L, and the total number of colonies reaches 10 ¹¹ cfu/L, the fermentation end point;

Among them, the mixed strain expansion liquid is added after mixing the Lactobacillus BL1 strain expansion liquid and the Lactobacillus BL2 strain expansion liquid by equal volume;

(4) Post-ripening and seasoning: after-ripening the product that has reached the end of fermentation at 4-8°C for 8-12 hours to facilitate the formation of aroma; pre-dissolve cyclamate and acesulfame potassium in 1 part by weight of water , 95 ° C for 20 minutes to sterilize, add to the fermented product in an aseptic operation and mix well.

(5) Packaging and storage: aseptically filled, circulated and stored at 4-8°C, shake well before drinking. The shelf life is 30 days. During the shelf life, the pH is ≥ 3.0, the total acid (calculated as lactic acid) is 5-9 g/L, and the total number of colonies is 10 ¹¹ cfu/L.

The preparation method of described Lactobacillus BL1 strain expansion liquid is:

(1) Lactobacillus BL1 probiotic activation: Streak culture of Lactobacillus BL1 on MRS solid medium, isolate a single colony and transfer to MRS slant medium, culture at 37°C for 24-72 hours for later use;

(2) Preparation of Lactobacillus BL1 liquid strain: inoculate the liquid MRS medium from a slant, and culture at 37°C for 18-24 hours to obtain a liquid strain;

(3) Preparation of Lactobacillus BL1 strain expansion solution: Prepare a bee pollen aqueous solution with a concentration of 3-5wt%, sterilize at 115°C for 15-20min, cool to room temperature, inoculate the liquid strain at a volume ratio of 3-5%, Static culture at 37°C for 1 to 2 days to obtain Lactobacillus BL1 strain expansion solution.

The preparation method of the Lactobacillus BL2 strain expansion liquid is the same as the preparation method of the Lactobacillus BL1 strain expansion liquid.

The present invention selects two excellent strains of Lactobacillus BL1 and Lactobacillus BL2, uses high-concentration bee pollen as the main raw material to carry out liquid mixed fermentation, and fully exerts the functions of the two bacteria in bee pollen fermentation. The present invention shows through research that Lactobacillus BL1 and Lactobacillus BL2 have strong acid resistance, and can utilize high-concentration bee pollen for rapid liquid fermentation. Lactobacillus BL2 produces acid quickly, but the acidification is more serious after fermenting bee pollen by a single bacterium, and the storage period is short. Lactobacillus BL1 produces acid slower than Lactobacillus BL2. The mixed fermentation of the two and the control of the fermentation process can ensure the number of probiotics within 30 days Keep it above 10 ¹¹ cfu/L. Lactobacillus BL1 has a strong ability to transform flavonoid glycosides, which can convert abundant flavonoid glycosides in bee pollen into corresponding aglycones, thereby improving the bioavailability of bee pollen by the human body. At the same time, Lactobacillus BL1 and Lactobacillus BL2 have antibacterial properties, and have certain inhibitory effects on food-borne pathogens such as Salmonella, Escherichia coli, Staphylococcus aureus and Vibrio parahaemolyticus; both strains have the ability to degrade cholesterol. The in vitro cholesterol degradation rates of Lactobacillus BL1 and Lactobacillus BL2 were 38.53% and 32.09%, respectively.

Generally speaking, the bee pollen active probiotic beverage provided by the present invention compounded and strengthened the health care functions of bee pollen and probiotics, and strengthened the anti-oxidation, anti-fatigue, anti-aging, improving immunity, and improving memory of bee pollen through biotransformation and other functions, and has a high content of probiotics, which can regulate gastrointestinal balance, lower cholesterol, integrate health care and treatment, cater to people's medical point of view of combining prevention, health care, treatment and rehabilitation, and have a shelf life of up to 30 days. Today, the market outlook is broader.

Description of drawings

Fig. 1 Degradation of main flavonoid glycosides in the fermentation process of bee pollen probiotics.

Detailed ways

The present invention will be further described below in conjunction with the examples, but not limited thereto.

Example 1:

Preparation of Lactobacillus BL1 strain expansion solution;

MRS solid medium is: peptone 5 g, beef extract 5 g, yeast extract 2. 5 g, glucose 2. 5 g, triammonium citrate 1. 0 g, sodium acetate 1. 56 g, potassium acetate 1. 125 g , disodium hydrogen phosphate 0. 815 g, manganese sulfate tetrahydrate 0. 125 g, magnesium sulfate heptahydrate 0. 29 g, Tween 80 is 0. 5 ml, agar 10 g, distilled water 500 ml, pH6. 5, 121 Sterilize at ℃ for 20 min.

MRS slant medium: same as MRS solid medium.

The liquid MRS medium is: remove the agar for the MRS solid medium components.

(1) Probiotic activation: Streak culture of Lactobacillus BL1 on MRS solid medium, isolate a single colony and transfer to MRS slant medium, culture at 37°C for 48 hours for later use;

(2) Liquid strains: Inoculate in liquid MRS medium from a slant and culture at 37°C for 20 hours to obtain liquid strains;

(3) Strain expansion solution: prepare 3% by weight bee pollen aqueous solution, sterilize at 115°C for 15 minutes, cool to room temperature, inoculate the liquid strain at a volume ratio of 4%, and culture at 37°C for 1 day to obtain Lactobacillus BL1 Strain expansion solution;

The preparation method of the Lactobacillus BL2 strain expansion solution is the same as above.

Example 2:

(1) Preparation of fermentation medium: remove impurities from bee pollen and crush through a 40-mesh sieve, and crush maltose and galactose through a 40-mesh sieve. Mix 5kg of bee pollen, 2kg of maltose and 2kg of galactose, slowly add 99kg of water while stirring, stir evenly and filter with gauze or 4 layers of gauze;

(2) Fermentation: The fermentation culture was sterilized by incubating at 115°C for 20 minutes, and cooled to below 37°C. Add 4% of the volume of the fermentation medium to the mixed strain expansion solution, where the mixed strain expansion solution is the two probiotic strain expansion solutions of Lactobacillus BL1 and Lactobacillus BL2 at a volume ratio of 1:1 and then added . 37 ℃ sealed static culture for 24 hours, when the pH of the fermentation broth drops to 3.10-3.20, the total acid (calculated as lactic acid) reaches 5-7 g/L, and the total number of colonies reaches 10 ¹¹ cfu/L, the fermentation end point;

(3) Post-ripening and seasoning: After-ripening the product that has reached the end of fermentation at 4°C for 12 hours to facilitate the formation of aroma. Dissolve 0.065kg of cyclamate and 0.030kg of acesulfame potassium in 1kg of water in advance, heat it at 95°C for 20 minutes to sterilize, add it to the fermented product in an aseptic operation and mix well;

(4) Packaging and storage: aseptically filled, circulated and stored at 4°C, shake well before drinking. The shelf life is 30 days. During the shelf life, pH≧3.0, total acid (calculated as lactic acid) 5-9 g/L, total bacterial colony 10 ¹¹ cfu/L.

The degradation of main flavonoid glycosides during the bee pollen probiotic fermentation process is shown in Figure 1 and Table 1.

It can be seen from Figure 1 that the main flavonoid glycosides kaempferol-3,4'-bis-O-β-D-glucoside and kaempferol-3-O-β-D-(2-O -β-D-glucosyl) glucopyranoside content decreased significantly. It can be seen from Table 1 that the content of kaempferol in bee pollen increased significantly after 1 day of fermentation, and the increase was 91%. In summary, after the mixed culture of two excellent strains of Lactobacillus BL1 and Lactobacillus BL2, the main flavonoid glycosides of bee pollen were partially biotransformed into the corresponding aglycon-kaempferol, which improved the bioavailability of bee pollen flavonoids.

Example 3

(1) Preparation of fermentation medium: remove impurities from bee pollen and crush through a 40-mesh sieve, and crush maltose through a 40-mesh sieve. Mix 6kg of bee pollen and 4kg of maltose, slowly add 99kg of water while stirring, stir evenly and filter with 4 layers of gauze;

(2) Fermentation: The fermentation culture was sterilized by incubating at 115°C for 25 minutes, and cooled to below 37°C. Add 4% of the volume of the fermentation medium to the mixed strain expansion solution, where the mixed strain expansion solution is the two probiotic strain expansion solutions of Lactobacillus BL1 and Lactobacillus BL2 at a volume ratio of 1:1 and then added . 37 ℃ sealed static culture for 24 hours, when the pH of the fermentation broth drops to 3.10-3.20, the total acid (calculated as lactic acid) reaches 5-7 g/L, and the total number of colonies reaches 10 ¹¹ cfu/L, the fermentation end point;

(3) Post-ripening and seasoning: After-ripening the product that has reached the end of fermentation at 4°C for 12 hours to facilitate the formation of aroma. Dissolve 0.05kg of cyclamate and 0.025kg of acesulfame potassium in 1kg of water in advance, heat it at 95°C for 25 minutes to sterilize, add it to the fermented product in an aseptic operation and mix well;

(4) Packaging and storage: aseptically filled, circulated and stored at 4°C, shake well before drinking. The shelf life is 30 days. During the shelf life, pH≧3.0, total acid (calculated as lactic acid) 5-9 g/L, total bacterial colony 10 ¹¹ cfu/L.

Example 4

(1) Preparation of fermentation medium: remove impurities from bee pollen and crush through a 40-mesh sieve, and crush galactose through a 40-mesh sieve. Mix 5kg of bee pollen and 3kg of galactose, slowly add 99kg of water while stirring, stir evenly and filter with gauze;

(2) Fermentation: The fermentation culture was sterilized by incubating at 115°C for 20 minutes, and cooled to below 37°C. Add 4% of the volume of the fermentation medium to the mixed strain expansion solution, where the mixed strain expansion solution is the two probiotic strain expansion solutions of Lactobacillus BL1 and Lactobacillus BL2 at a volume ratio of 1:1 and then added . 37 ℃ sealed static culture for 24 hours, when the pH of the fermentation broth drops to 3.10-3.20, the total acid (calculated as lactic acid) reaches 5-7 g/L, and the total number of colonies reaches 10 ¹¹ cfu/L, the fermentation end point;

(3) Post-ripening and seasoning: After-ripening the product that has reached the end of fermentation at 4°C for 12 hours to facilitate the formation of aroma. Dissolve 0.04kg of cyclamate and 0.025kg of acesulfame potassium in 1kg of water in advance, sterilize at 95°C for 20 minutes, add them to the fermented product in an aseptic manner and mix well;

(4) Packaging and storage: aseptically filled, circulated and stored at 4°C, shake well before drinking. The shelf life is 30 days. During the shelf life, pH≧3.0, total acid (calculated as lactic acid) 5-9 g/L, total bacterial colony 10 ¹¹ cfu/L.

Claims (6)

1. prepare a method for pollen active probiotic drink, it is characterized in that,

(1) raw material is got by following weight parts: Bee Pollen 3 ~ 10, maltose and/or galactolipin 3 ~ 5, water 100, honey element 0.030 ~ 0.065, acesulfame potassium 0.020 ~ 0.030;

(2) preparation of fermentation medium: add water by after Bee Pollen and maltose and/or galactolipin mixing, stirs and by gauze or 4 layers of filtered through gauze;

(3) ferment: by fermentation medium high temperature sterilization, and be cooled to less than 37 DEG C; Then the mixed bacteria adding fermentation medium volume 3 ~ 5% expands liquid, and 30 ~ 37 DEG C of sealing and standing are cultivated; Treat that zymotic fluid pH is down to 3.10 ~ 3.30, total acid reaches 5 ~ 7 g/L, and total plate count reaches 10 ¹¹it is fermentation termination during cfu/L;

Wherein mixed bacteria expands liquid is expand liquid with lactobacillus BL1 bacterial classification expansion liquid and lactobacillus BL2 bacterial classification to add by after equal-volume mixing;

(4) after-ripening and seasoning: the product reaching fermentation termination is left standstill in 4 ~ 8 DEG C and carries out after-ripening in 8 ~ 12 hours; By honey element and acesulfame potassium water-soluble solution in advance, high temperature sterilization, to join in fermented product in sterile working mode and mixes rear sterile filling.

2. prepare the method for pollen active probiotic drink as claimed in claim 1, it is characterized in that, the preparation method that described lactobacillus BL1 bacterial classification expands liquid is:

(1) lactobacillus BL1 probio activation: cultivations of rule on MRS solid medium by lactobacillus BL1, is separated single bacterium colony and transfers MRS slant medium, and 37 DEG C of cultivations 24 ~ 72 hours are for subsequent use;

(2) preparation of lactobacillus BL1 liquid spawn: by inclined plane inoculating in liquid MRS culture medium, cultivates 18 ~ 24 hours, obtains liquid spawn for 37 DEG C;

(3) lactobacillus BL1 bacterial classification expands the preparation of liquid: compound concentration is the Bee Pollen aqueous solution of 3 ~ 5wt%, 115 DEG C of sterilizing 15 ~ 20min, are cooled to normal temperature, with the volume ratio inoculation liquid spawn of 3 ~ 5%, 37 DEG C of quiescent culture 1 ~ 2 day, obtain lactobacillus BL1 bacterial classification and expand liquid;

Described lactobacillus BL2 bacterial classification expands the preparation method of preparation method with lactobacillus BL1 bacterial classification expansion liquid of liquid.

3. prepare the method for pollen active probiotic drink as claimed in claim 1 or 2, it is characterized in that, the inoculation volume ratio that described mixed bacteria expands liquid is 4%.

4. prepare the method for pollen active probiotic drink as claimed in claim 1 or 2, it is characterized in that, described Bee Pollen removal of impurities also pulverized 40 ~ 80 mesh sieves, and 40 mesh sieves pulverized by maltose and/or galactolipin.

5. prepare the method for pollen active probiotic drink as claimed in claim 1 or 2, it is characterized in that, described fermented and cultured carries out high temperature sterilization in 15 ~ 30 minutes based on 115 DEG C of insulations.

6. prepare the method for pollen active probiotic drink as claimed in claim 1 or 2, it is characterized in that, described by after honey element and the dissolving of acesulfame potassium water in advance, within 20 minutes, carry out high temperature sterilization in 95 DEG C of insulations.