Solid beverage capable of adsorbing heavy metal ions and preparation method thereof

Abstract

The invention relates to the technical field of microorganisms, in particular to a solid beverage capable of adsorbing heavy metal ions and a preparation method of the solid beverage. The preparation method provided by the invention comprises the following steps: adding bifidobacterium longum subsp. Longum BD3150 into skimmed milk, uniformly mixing, and drying to obtain the solid beverage. The lead adsorption rate of the bifidobacterium longum solid beverage is larger than or equal to 60%, the lead adsorption capacity of thalli in unit weight is larger than 30 mg / g of the weight of the thalli, and after the bifidobacterium longum solid beverage is preserved for 6 months at normal temperature, the lead and copper adsorption rate of the solid beverage can be kept stable.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, in particular to a solid beverage capable of absorbing heavy metal ions and a preparation method thereof. Background Art

[0002] Currently, chelation therapy is commonly used to treat heavy metal poisoning, which uses chelators to accelerate the excretion of metal ions. For example, when treating lead poisoning, commonly used drugs include ethylenediaminetetraacetic acid (EDTA) and dimercaptosuccinic acid (DMSA). However, these chelators may cause some adverse reactions and are not suitable for high-dose or long-term treatment.

[0003] Compared with chelating agents, the use of strains that can adsorb heavy metal ions and adapt to the gastrointestinal environment for the treatment of heavy metal poisoning can effectively avoid the adverse reactions of chelating agent therapy. Summary of the invention

[0004] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a solid beverage capable of adsorbing heavy metal ions and a preparation method thereof, so as to solve the problems in the prior art.

[0005] To achieve the above object and other related objects, the present invention provides a method for preparing a solid beverage capable of absorbing heavy metal ions, the preparation method comprising the following steps: adding Bifidobacterium longum subspecies longum BD3150 into skim milk and mixing, and obtaining a solid beverage after drying.

[0006] Preferably, based on the treatment of solid beverage, the number of viable bacteria of Bifidobacterium longum subspecies longum BD3150 in the solid beverage is 1×10 8 -1x10 10 CFU / g.

[0007] The present invention also provides use of a solid beverage in preparing a heavy metal ion adsorption product. The solid beverage contains Bifidobacterium longum subspecies longum BD3150.

[0008] The present invention also provides a solid beverage capable of absorbing heavy metal ions. The solid beverage contains Bifidobacterium longum subspecies longum BD3150 and skim milk.

[0009] As described above, the solid beverage capable of absorbing heavy metal ions and the preparation method thereof of the present invention have the following beneficial effects:

[0010] The adsorption rate of lead by the Bifidobacterium longum solid beverage provided by the present invention is ≥60%, the lead adsorption amount per unit weight of the bacteria is >30 mg / g bacterial weight, and after being stored at room temperature for 6 months, the adsorption rate of lead and copper by the solid beverage of the present invention can also remain stable. DETAILED DESCRIPTION

[0011] The invention provides use of a solid beverage in preparing a heavy metal ion adsorption product. The solid beverage contains Bifidobacterium longum subsp. longum BD3150.

[0012] In the present invention, Bifidobacterium longum subsp. longum BD3150 is deposited in the General Microbiology Center CGMCC of China Microorganism Culture Collection Administration, with a deposit number of CGMCC No.20105, a deposit date of 2020-06-18, and a deposit address of Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.

[0013] In some specific embodiments, the heavy metal ion is selected from one or more of copper ion, lead ion, zinc ion, tin ion, nickel ion, cobalt ion, antimony ion, mercury ion, cadmium ion or bismuth ion. Preferably, the heavy metal ion is lead ion.

[0014] The invention also provides a solid beverage capable of absorbing heavy metal ions. The solid beverage contains Bifidobacterium longum subsp. longum BD3150 and skim milk.

[0015] In some specific embodiments, based on the mass of the solid beverage, the viable count of Bifidobacterium longum subsp. longum BD3150 in the solid beverage is 1×10 10 -1x10 11 CFU / g. Specifically, the viable count is 1x10 10 -2.1x10 10 CFU / g, 2.1x10 10 -2.7x10 10 CFU / g, 2.7x10 10 -3.9x10 9 CFU / g, 3.9x10 10 -4.8x10 10 CFU / g, 4.8x10 10 -5x10 10 CFU / g, 5x10 10 -6.5x10 10 CFU / g, 6.5x10 10 -8.5x10 10 CFU / g or 8.5x1010 -1x10 11 CFU / g.

[0016] In some specific embodiments, the solid beverage further contains one or more of a sweetener, a vitamin, a mineral, an edible flavor, an edible coloring, a preservative, a stabilizer, a thickener or an acidity regulator.

[0017] In some specific embodiments, the sweetener can be selected from one or more of sucrose, fructose, glucose, erythritol, xylitol, corn syrup or fructose syrup.

[0018] In some specific embodiments, the vitamin can be selected from one or more of vitamin C, vitamin A, vitamin D, vitamin E, or vitamin B1, B2, B3, B5, B6, B7, B9, and B12.

[0019] In some specific embodiments, the mineral may be selected from one or more of sodium salts, potassium salts, magnesium salts, calcium salts or phosphates.

[0020] In some embodiments, the preservative can be selected from one or more of benzoic acid or its salts, sorbic acid or its salts, p-hydroxybenzoic acid esters or its salts, or dehydroacetic acid or its salts.

[0021] In some specific embodiments, the stabilizer or thickener can be selected from one or more of pectin, sodium carboxymethyl cellulose, xanthan gum, sodium alginate, carrageenan, gellan gum, monoglyceride, sucrose ester or locust bean gum.

[0022] In some embodiments, the acidity regulator can be selected from one or more of citric acid or its salts, malic acid or its salts, gluconic acid or its salts, or tartaric acid or its salts.

[0023] The present invention also provides a method for preparing the solid beverage, the method comprising the following steps:

[0024] Bifidobacterium longum subsp. longum BD3150 is added into skim milk and mixed evenly, and then dried to obtain a solid beverage.

[0025] In some embodiments, based on the volume of skim milk after mixing, the viable count of Bifidobacterium longum subsp. longum BD3150 is 1×10 9 -1x10 10 CFU / mL. Specifically, the viable count is 1x10 9 -1.2x109 CFU / mL, 1.2x10 9 -1.68x10 9 CFU / mL, 1.68x10 9 -2.7x10 9 CFU / mL, 2.7x10 9 -2.88x10 9 CFU / mL, 2.88x10 9 -5x10 9 CFU / mL, 5x10 9 -7.5x10 9 CFU / mL or 7.5x10 9 -1x10 10 CFU / mL.

[0026] In some embodiments, the skim milk comprises skim milk powder and water.

[0027] Furthermore, based on the mass of skim milk, the mass percentage of skim milk powder to the total mass of skim milk is 5-15%. Specifically, the mass percentage is 5-6%, 6-7%, 7-8%, 8-9%, 9-10%, 10-11%, 11-12%, 12-13%, 13-14% or 14-15%.

[0028] Further, based on the mass of skim milk, the mass percentage of water in the total mass of skim milk is 85-95%. Specifically, the mass percentage is 85-86%, 86-87%, 87-88%, 88-89%, 89-90%, 90-91%, 91-92%, 92-93%, 93-94% or 94-95%.

[0029] In the present invention, "fixed beverage" generally refers to a solid product made from sugar, milk and dairy products, eggs or egg products, fruit juice or edible plant extracts as main raw materials, with appropriate amount of auxiliary materials or food additives added, and the water content of each 100 grams of the finished product is not higher than 5 grams.

[0030] In some embodiments, the drying method can be selected from one or more of freeze drying, vacuum drying or fluidized bed drying.

[0031] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0032] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing the specific embodiments rather than for limiting the scope of protection of the present invention; in the present specification and claims, unless otherwise expressly stated herein, the singular forms "a", "an" and "the" include plural forms.

[0033] When the embodiments give numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, according to the grasp of the prior art by those skilled in the art and the record of the present invention, any methods, equipment, and materials of the prior art similar or equivalent to the methods, equipment, and materials described in the embodiments of the present invention can also be used to realize the present invention.

[0034] Example 1

[0035] 1. Materials and methods

[0036] (a) Preparation of Bifidobacterium longum BD3150 bacterial agent

[0037] The lyophilized powder of Bifidobacterium longum BD3150 was dissolved with a small amount of sterile distilled water, and a ring was streaked on TPY solid medium (purchased from Qingdao High-tech Industrial Park Haibo Biotechnology Co., Ltd., China) with an inoculation loop, and anaerobically cultured at 37°C for 48 hours. A single colony was picked up with an inoculation loop and placed in 10 mL TPY liquid medium (purchased from Qingdao High-tech Industrial Park Haibo Biotechnology Co., Ltd., China). The colony was evenly dispersed in the liquid medium using a vortex oscillator, cultured at 37°C for 24 hours, and inoculated into TPY liquid medium with an inoculum of 2% (v / v). After another 24 hours of culture at 37°C, the culture was centrifuged at 15,000 rpm for 10 minutes, the supernatant was discarded, the bacteria were washed twice with sterile distilled water, and then suspended with a small amount of skim milk (10%, v / w), and freeze-dried to obtain the bacterial agent of Bifidobacterium longum BD3150. The concentration of the bacterial agent was determined to be 2×10 11 CFU / g.

[0038] (b) Preparation of skim milk:

[0039] Mix 10% skim milk powder by mass with 90% water, and after fully dissolving, sterilize at 90° C. for 10 minutes and cool to room temperature to obtain sterile skim milk of the required concentration.

[0040] 2. Preparation of Bifidobacterium longum BD3150 solid beverage

[0041] Bifidobacterium longum BD3150 was added to a final concentration of 2.7 x 10 9 CFU / mL was added to the sterile skim milk, and freeze-dried to obtain the long bifidobacterium solid beverage A of this embodiment. After testing, the viable count of the long bifidobacterium solid beverage A was 2.7x10 10 CFU / g.

[0042] Example 2

[0043] 1. Materials and methods

[0044] (a) Preparation of Bifidobacterium longum BD3150 bacterial agent: same as Example 1.

[0045] (b) Preparation of skim milk:

[0046] 12% skim milk powder by mass was mixed with 88% water, and after fully dissolved, sterilized at 135° C. for 5 seconds and cooled to room temperature to obtain sterile skim milk of the required concentration.

[0047] 2. Preparation of Bifidobacterium longum BD3150 solid beverage

[0048] Bifidobacterium longum BD3150 was added to a final concentration of 1.2 x 10 9 CFU / mL was added to the sterile skim milk, and freeze-dried to obtain the long bifidobacterium solid beverage B of this embodiment. After testing, the viable count of the long bifidobacterium solid beverage B was 1x10 10 CFU / g.

[0049] Example 3

[0050] 1. Materials and methods

[0051] (a) Preparation of Bifidobacterium longum BD3150 bacterial agent: same as Example 1.

[0052] (b) Preparation of skim milk:

[0053] Mix 6% skim milk powder with 94% water, and after fully dissolving, sterilize at 90°C for 10 minutes and cool to room temperature to obtain sterile skim milk of the required concentration.

[0054] 2. Preparation of Bifidobacterium longum BD3150 solid beverage

[0055] Bifidobacterium longum BD3150 was added to a final concentration of 2.88 x 10 9 CFU / mL was added to the sterile skim milk, and freeze-dried to obtain the long bifidobacterium solid beverage C of this embodiment. After testing, the viable count of the long bifidobacterium solid beverage C was 4.8x1010 CFU / g.

[0056] Example 4

[0057] 1. Materials and methods

[0058] (a) Preparation of Bifidobacterium longum BD3150 bacterial agent: same as Example 1.

[0059] (b) Preparation of skim milk:

[0060] Mix 8% skim milk powder by mass with 92% water, sterilize at 135°C for 5s after fully dissolving, and cool to room temperature to obtain sterile skim milk of the required concentration.

[0061] 2. Preparation of Bifidobacterium longum BD3150 solid beverage

[0062] Bifidobacterium longum BD3150 was added to a final concentration of 1.68 x 10 9 CFU / mL was added to the sterile skim milk, and freeze-dried to obtain the long bifidobacterium solid beverage D of this embodiment. After testing, the viable count of the long bifidobacterium solid beverage D was 2.1x10 10 CFU / g.

[0063] Example 5

[0064] 1. Materials and methods

[0065] (a) Preparation of Bifidobacterium longum BD3150 bacterial agent: same as Example 1.

[0066] (b) Preparation of skim milk:

[0067] Mix 7% skim milk powder with 93% water, and after fully dissolving, sterilize at 90°C for 10 minutes and cool to room temperature to obtain sterile skim milk of the required concentration.

[0068] 2. Preparation of Bifidobacterium longum BD3150 solid beverage

[0069] Bifidobacterium longum BD3150 was added to a final concentration of 2.73 x 10 9 CFU / mL was added to the sterile skim milk, and freeze-dried to obtain the long bifidobacterium solid beverage E of this embodiment. After testing, the viable count of the long bifidobacterium solid beverage E was 3.9x10 10 CFU / g.

[0070] Example 6 Heavy metal adsorption effect of Bifidobacterium longum solid beverage

[0071] Take 1 g of each of the Bifidobacterium longum solid beverages A to E of Examples 1 to 5 above, dissolve them in 10 mL of a lead or copper ion aqueous solution (20 mg / L), shake at 37°C and 200 rpm for 1 h, centrifuge at 6,000 rpm for 15 min, and collect the supernatant solution and bacterial precipitate.

[0072] After the supernatant solution was filtered through a 0.22 μm filter membrane, the concentration of lead or copper ions in the solution was detected by inductively coupled plasma optical emission spectrometry (ICP-OES). The removal rate was calculated as follows:

[0073]

[0074] In the above formula, C 初始 is the metal solubility in the original solution before adsorption (mg / L); C 实验 is the metal solubility in the supernatant after adsorption (mg / L).

[0075] After freeze-drying, the bacterial precipitate was added with 2.5 mL of concentrated nitric acid (super pure) and digested in a graphite digester at 130°C for 30 min. The tube was rinsed twice with deionized water and the volume was fixed to 10 mL. The concentration of lead or copper ions in the solution was detected by inductively coupled plasma optical emission spectrometry (ICP-OES). The heavy metal adsorption per unit weight of the bacteria was obtained by conversion. The results are shown in the following table.

[0076] Table 1 Heavy metal removal rate of Bifidobacterium longum solid beverages in Examples 1 to 5 and heavy metal adsorption per unit weight of bacteria

[0077] Bifidobacterium longum solid beverage Comparison A B C D E Lead removal rate in supernatant (%) 50 77 60 90 69 85 Lead adsorption (mg / g) 25 38.5 30 45 34.5 42.5

[0078] It can be seen from the above table that although the control group (sterile skim milk powder) itself has a certain adsorption effect on lead, when Bifidobacterium longum BD3150 is added to the solid beverage, the removal rate (adsorption rate) of lead is further improved. Specifically, the removal rate (adsorption rate) of Bifidobacterium longum solid beverage for lead is ≥60%, and the adsorption amount of lead is ≥30 mg / g.

[0079] Example 6 Experiment on storage of Bifidobacterium longum solid beverage at room temperature

[0080] The Bifidobacterium longum solid beverages A to E of Examples 1 to 5 were stored at room temperature (25°C) and taken out at 0, 3 and 6 months. The removal rates of lead and copper of each sample were measured by referring to the method described in Effect Example 1. The results are shown in the following table.

[0081] Table 2 Removal rate of heavy metals by Bifidobacterium longum solid beverages A to E during storage at room temperature (25°C)

[0082]

[0083]

[0084] It can be seen from the above table that the lead adsorption rate of the Bifidobacterium longum solid beverage prepared by the present invention remains stable after being stored at room temperature (25° C.) for 6 months.

[0085] Example 7 Comparative experiment on the removal of heavy metal ions from Bifidobacterium longum solid beverage

[0086] Comparison of different bacterial concentrations

[0087] The final concentration of the Bifidobacterium longum BD3150 bacterial agent added in Example 1 was adjusted to obtain the following group of Bifidobacterium longum solid beverages prepared by different methods. The removal rates of lead and copper by the solid beverages were measured by the method described in Effect Example 1, and the results are shown in the following table.

[0088] Table 3 Removal rate of lead by Bifidobacterium longum solid beverage prepared by different methods

[0089] Comparison plan 1 Final concentration of bacterial agent added (CFU / mL) <![CDATA[5x10 8 ]]> Lead removal rate of solid beverages (%) 54

[0090] From the results shown in the above table, it can be seen that when the final concentration of the Bifidobacterium longum BD3150 bacterial agent added in the preparation method of the Bifidobacterium longum solid beverage is adjusted to outside the preferred range, the removal (adsorption) effect of the prepared solid beverage on lead still exists, but compared with the removal effect of the control sample (sterile skim milk) described in Example 1 (removal rate 50%), the solid beverage prepared by adding this dose of Bifidobacterium longum has a relatively limited effect on improving the removal rate (adsorption rate) of heavy metals.

[0091] Comparison of different bacterial species

[0092] Referring to the method described in Example 1, the heavy metal removal rate and the heavy metal adsorption per unit weight of the solid beverages prepared by adding Bifidobacterium longum BD3150 (provided by Bright Dairy Co., Ltd., Shanghai, China) and Bifidobacterium animalis BB12 (provided by Chr. Hansen, Denmark) were compared. The specific operation is as follows.

[0093] 1. Materials and methods

[0094] (a) Preparation of Bifidobacterium longum BD3150 bacterial agent: same as Example 1.

[0095] (b) Preparation of Bifidobacterium animalis BB12 bacterial agent

[0096] The freeze-dried powder of animal Bifidobacterium BB12 was dissolved with a small amount of sterile distilled water, and a ring was streaked on TPY solid medium (purchased from Qingdao High-Tech Industrial Park Haibo Biotechnology Co., Ltd., China) with an inoculation loop, and anaerobically cultured at 37°C for 48 hours. A single colony was picked up with an inoculation loop and placed in 10 mL TPY liquid medium (purchased from Qingdao High-Tech Industrial Park Haibo Biotechnology Co., Ltd., China). The colony was evenly dispersed in the liquid medium using a vortex oscillator, cultured at 37°C for 24 hours, and inoculated into TPY liquid medium with an inoculum of 2% (v / v). After culturing again at 37°C for 24 hours, the culture was centrifuged at 15,000 rpm for 10 minutes, the supernatant was discarded, the bacteria were washed twice with sterile distilled water, and then suspended with a small amount of skim milk (10%, v / w), and freeze-dried to obtain the bacterial agent of Bifidobacterium BB12. The concentration of the bacterial agent was determined to be 1.5×10 11 CFU / g.

[0097] Preparation of skim milk: same as in Example 1.

[0098] 2. Preparation of Bifidobacterium Solid Beverage

[0099] Bifidobacterium longum BD3150 and Bifidobacterium animalis BB12 were added to a final concentration of 2.7 × 10 9 CFU / mL were added to the above sterile skim milk respectively, and then freeze-dried to obtain solid beverages prepared by adding different bifidobacteria.

[0100] 3. Comparison of heavy metal adsorption effects

[0101] The heavy metal removal rate of the solid beverage and the heavy metal adsorption per unit weight of the bacteria were measured by the method described in Reference Example 1. The results are shown in the following table.

[0102] Table 4 Heavy metal removal rate of different bifidobacterium solid beverages and heavy metal adsorption per unit weight of bacteria

[0103] Bifidobacterium solid beverage Comparison BD3150 BB12 Lead removal rate in supernatant (%) 50 77 52 Lead adsorption (mg / g) 25 38.5 26

[0104] As can be seen from the above table, the lead removal effect of the solid beverage containing Bifidobacterium BB12 is basically the same as that of the control group, indicating that Bifidobacterium BB12 does not have the effect of promoting the adsorption of heavy metals by solid beverages, while the lead removal rate (adsorption rate) of the solid beverage containing Bifidobacterium longum BD3150 is significantly higher than that of the control sample and the solid beverage containing the same number of bacteria BB12, indicating that Bifidobacterium longum BD3150 itself has a certain heavy metal adsorption effect, and it adsorbs lead in the environment together with skim milk. This superposition effect further enhances the final adsorption effect.

[0105] The above examples are for the purpose of illustrating the embodiments disclosed by the present invention and are not to be construed as limiting the present invention. In addition, the various modifications listed herein and the variations of the methods in the invention are obvious to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been specifically described in conjunction with various specific preferred embodiments of the present invention, it should be understood that the present invention should not be limited to these specific embodiments. In fact, various modifications as described above that are obvious to those skilled in the art to obtain the invention should be included within the scope of the present invention.

Claims

1. A solid beverage capable of absorbing heavy metal ions, characterized in that: The solid beverage contains Bifidobacterium longum subsp. longum BD3150 and skimmed milk powder, wherein the preservation number of Bifidobacterium longum subsp. longum BD3150 is CGMCC No.20105.

2. The solid beverage according to claim 1, characterized in that Based on the quality of the solid beverage, the viable count of Bifidobacterium longum subsp. longum BD3150 in the solid beverage is 1×10 10 -1x10 11 CFU / g.

3. The solid beverage according to claim 1, characterized in that The solid beverage further contains one or more of a sweetener, vitamins, minerals, edible flavors, edible colorings, preservatives, stabilizers, thickeners or acidity regulators.

4. The solid beverage according to claim 3, characterized in that: The solid beverage comprises one or more of the following features: 1) The sweetener is selected from one or more of sucrose, fructose, glucose, erythritol, xylitol, corn syrup or fructose syrup; 2) The vitamin is selected from one or more of vitamin C, vitamin A, vitamin D, vitamin E, or vitamin B1, B2, B3, B5, B6, B7, B9, and B12; 3) The mineral is selected from one or more of sodium salt, potassium salt, magnesium salt, calcium salt or phosphate; 4) The preservative is selected from one or more of benzoic acid or its salts, sorbic acid or its salts, p-hydroxybenzoic acid esters or its salts, or dehydroacetic acid or its salts; 5) The stabilizer or thickener is selected from one or more of pectin, sodium carboxymethyl cellulose, xanthan gum, sodium alginate, carrageenan, gellan gum, monoglyceride, sucrose ester or locust bean gum; 6) The acidity regulator is selected from one or more of citric acid or its salts, malic acid or its salts, gluconic acid or its salts, or tartaric acid or its salts.

5. Use of the solid beverage according to any one of claims 1 to 4 in the preparation of heavy metal ion adsorption products.

6. The use according to claim 5, characterized in that The heavy metal ions are selected from one or more of copper ions, lead ions, zinc ions, tin ions, nickel ions, cobalt ions, antimony ions, mercury ions, cadmium ions or bismuth ions.

7. The method for preparing the solid beverage according to claim 1 or 2, characterized in that: The preparation method comprises the following steps: adding Bifidobacterium longum subsp. longum BD3150 into skim milk and mixing evenly, and then drying to obtain a solid beverage.

8. The preparation method according to claim 7, characterized in that: Based on the volume of skim milk after mixing, the number of viable bacteria of Bifidobacterium longum subsp. longum BD3150 is 1x10 9 -1x10 10 CFU / mL.

9. The preparation method according to claim 7, characterized in that: The skim milk contains skim milk powder and water; preferably, based on the mass of the skim milk, the mass percentage of the skim milk powder is 5-15% of the total mass of the skim milk; and / or, based on the mass of the skim milk, the mass percentage of water is 85-95% of the total mass of the skim milk.

10. The preparation method according to claim 7, characterized in that: The drying method is selected from one or more of freeze drying, vacuum drying or fluidized bed drying.

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