CN100497594C - A compound protective agent for high activity bifidobacterium dry powder preparation - Google Patents

Abstract

The invention discloses a composite protective agent of a high-activity bifidobacterium dry powder preparation, which comprises the following components in percentage by weight: 10g of beta-cyclodextrin, 2-5g of lactose, 0.5-2g of trehalose, 0.5-2g of yeast extract, 0.05-0.1g of vitamin E and 100mL of distilled water. The freeze dried bifidobacterium powder prepared with the bifidobacterium powder has live bacteria content as high as 1.0 x 10¹¹cfu/g or more; sealing under normal pressure, storing at 4 deg.C for 1 year, and the viable bacteria content is reduced by less than 1 log order of magnitude, and is 10¹⁰More than cfu/g, solves the key technical problems in the process of developing the high-activity bifidobacterium dry powder preparation. The preparation process is simple and convenient. The freeze dried Bifidobacterium bifidum or Bifidobacterium longum powder prepared with the efficient composite protectant has high live bacteria content, long preservation period, convenient preservation and transportation and easy industrial production, and may be further prepared into capsule, tablet, powder and other product forms, and the freeze dried Bifidobacterium bifidum or Bifidobacterium longum powder may be taken orally directly and used as additive and leaven for developing other functional Bifidobacterium food and medicine preparation.

Description

A compound protective agent for high activity bifidobacterium dry powder preparation

technical field

The invention relates to a compound protective agent for a high-activity bifidobacterium dry powder preparation, and belongs to the technical field of bifidobacterium product preparation.

Background technique

Bifidobacteria (Bifidobacteria) are typical beneficial bacteria in the human intestinal tract, and the number of Bifidobacteria in the human intestinal tract has been increasingly valued as an important indicator to measure the health of the body. However, due to the aging of the body, environmental pollution, dietary changes, abuse of antibiotics, radiotherapy and chemotherapy, etc., the number of bifidobacteria in the human intestinal tract often decreases, leading to intestinal microecological imbalance, causing bacterial translocation, inducing endogenous infection, and causing Body disease. Therefore, the research and development of bifidobacterium products, supplementing the number of bifidobacteria colonized in the human intestinal tract through exogenous means, is of great significance for adjusting the microecological balance of the human intestinal tract, improving immunity, and promoting health. In recent years, the industry of bifidobacterium products at home and abroad has developed rapidly, becoming one of the fastest growing industries in the world, occupying an increasingly important position in the food and pharmaceutical industries, with more than 70 products. However, because bifidobacteria have special biological characteristics such as strict nutritional requirements, obligate anaerobic, and sensitivity to low pH environments, many bifidobacteria products currently on the market generally have low viable bacteria content and short shelf life. Quality issues. Especially for liquid products, under refrigerated conditions, the preservation period of live bacteria (viable bacteria content ≥ 1×10 ⁶ cfu/mL) is generally only 5-7 days. Therefore, improving the viable content of bifidobacterium products and prolonging the storage period of viable bacteria are key technical issues to be solved urgently in the research and development of bifidobacterium viable products. In order to increase the live bacteria content of Bifidobacterium liquid products and prolong the preservation period of live bacteria, researchers at home and abroad have successively adopted the technology of increasing the pH value of the substrate, obtaining artificially domesticated oxygen-resistant strains and acid-resistant strains, and combined with refrigeration and freezing preservation methods. . Under refrigerated conditions, the preservation period of liquid live bacteria (live bacteria content ≥ 1×10 ⁶ cfu/mL) can be extended up to 15d-21d by adopting the technology of increasing the pH value of the substrate and obtaining artificially domesticated oxygen-resistant and acid-resistant strains ; And adopt the method for cryopreservation, although can prolong the preservation period of liquid live bacteria significantly, maintain freezing expense higher, transportation is inconvenient, and the dependence on cryopreservation center is too strong. Bifidobacterium dry powder products are produced by adopting excellent strains of Bifidobacteria through liquid deep-layer high-density culture, concentration and separation, miscibility with protective agent medium, drying process, and aseptic packaging. The drying process includes: vacuum low-temperature drying method, spray drying method and vacuum freeze-drying method (abbreviation: freeze-drying method). Bifidobacteria dry powder products prepared by vacuum low-temperature drying method and spray drying method generally have relatively low viable bacteria content; while bifidobacterium dry powder products prepared by freeze-drying technology not only have high viable bacteria content and long storage period, but also can be stored and The transportation is convenient, the degree of mechanization is high, and it is convenient for industrialized mass production; it can be directly taken orally, and can also be used as an additive and a starter for the research and development of other bifidobacteria functional foods and pharmaceutical preparations, and has extremely broad prospects for promotion. The main factors that affect the content of viable bacteria in freeze-dried products of Bifidobacterium (cell freeze-dried survival rate) and the preservation period of viable bacteria (cell storage stability) are: strain characteristics, medium, culture conditions, harvest period, initial cell concentration, Lyoprotectant and its pH value, freezing speed, drying process, residual moisture, packaging form, storage temperature, etc. Among them, the lyoprotectant is the most complicated and difficult key factor to choose. It not only affects the cell survival rate of Bifidobacteria during the freeze-drying process, but also affects the cell stability during preservation. Therefore, the formulation of the lyoprotectant and its preparation method are commercial secrets.

Contents of the invention

The object of the present invention is to provide a compound protective agent for high-activity bifidobacterium dry powder preparations, which solves the problems of low viable bacteria content and short storage period in the production process of bifidobacteria dry powder products and their functional foods and pharmaceutical preparations. .

The idea of the present invention is like this. The inventor used self-separated and bred bifidobacteria (Bifidobacterium bifidum Bbm and Bifidobacterium longum B1m) with high vitality, strong oxygen resistance and stress resistance as test strains, and studied nineteen kinds of food-grade single protective bacteria. The effects of different agents on the freeze-drying survival rate of Bifidobacterium cells were preliminarily screened for a single protective agent against freezing or drying. On this basis, the effects of different compound protective agents on the freeze-drying survival rate of Bifidobacterium cells were studied, and five excellent anti-freeze-drying compound protectants were screened and optimized. The cell acceleration test was carried out on the freeze-dried powder of bifidobacteria made from five kinds of composite protective agents, and an excellent composite protective agent that was resistant to freeze-drying and storage was further screened out. Specifically, the formula of the compound protective agent of the highly active bifidobacterium dry powder preparation in the present invention consists of: β-cyclodextrin 10g, lactose 2-5g, trehalose 0.5-2g, yeast extract 0.5-2g, vitamin E 0.05-0.1g, distilled water 100mL.

Composite protecting agent preparation technology of the present invention is:

① Weigh β-cyclodextrin according to the amount, dry heat sterilization at 105°C for 60 minutes, and cool;

②Weigh the vitamin E according to the amount, filter and sterilize;

③Adjust the pH value of the distilled water used to prepare the compound protective agent with lye in advance to about 7.0;

④ Add lactose, trehalose and yeast extract in the distilled water with pH adjusted in advance, heat to dissolve and mix well;

⑤ Sterilize at 115°C for 10 minutes and cool to room temperature;

⑥ Add β-cyclodextrin and vitamin E, and mix well.

The bifidobacteria freeze-dried powder prepared by using the composite protective agent of the present invention is packaged in different storage methods (atmospheric pressure storage, vacuum storage), stored at different temperatures (-18°C, 4°C), and regularly tested for freeze-drying The number of viable bacteria in the powder was determined, and the storage conditions and storage period of the freeze-dried bifidobacterium powder prepared by the composite protective agent were determined, so as to directly investigate the actual application effect of the composite protective agent. The research results show that: the freeze-dried powder of single strain of Bifidobacterium bifidum Bbm or Bifidobacterium longum Blm made by using this composite protective agent has a live bacteria content of more than 1.0×10 ¹¹ cfu/g; The freeze-dried powder of single strains of Bifidobacterium longum Blm and Bifidobacterium bifidum Bbm prepared by the formula is preserved for 1 year under the same storage method at different storage temperatures or at the same storage temperature and different storage methods, and the content of viable bacteria decreases by less than 1 log order of magnitude, all above 10 ¹⁰ cfu/g.

The technical effect that the present invention obtains is:

(1) The high-efficiency composite protective agent raw materials for preparing the single strain freeze-dried powder of Bifidobacterium longum and Bifidobacterium bifidum provided by the present invention are easy to get and low in cost;

(2) The composite protecting agent of the present invention has simple process and convenient manufacture;

(3) The bifidobacteria freeze-dried powder prepared by the high-efficiency composite protective agent of the present invention solves the key technical problems in the process of researching and developing the high-activity bifidobacteria dry powder preparation.

(4) The Bifidobacterium bifidum or Bifidobacterium longum freeze-dried powder prepared by the high-efficiency composite protective agent of the present invention not only has high live bacteria content, long storage period, convenient storage and transportation, and is convenient for industrialized batch production, but also can According to needs, it can be made into different product forms such as capsules, tablets, powders, etc., which can be directly taken orally, and can also be used as additives and starters for the development of other bifidobacteria functional foods and pharmaceutical preparations.

Detailed ways

The following examples serve to illustrate the invention.

Example 1

1. The compound protective agent formula of the present invention: β cyclodextrin 10g, lactose 5g, trehalose 1g, yeast extract 1g, vitamin E 0.1g, water 100mL (adjust the pH value of distilled water to about 7.0 with lye in advance).

2. Preparation:

Accurately weigh and mix the components (except β-cyclodextrin and vitamin E) that make up the composite protective agent; heat to dissolve and cool to room temperature; sterilize at 115°C for 10 minutes and cool to room temperature; add aseptically Filter-sterilized vitamin E and β-cyclodextrin sterilized by dry heat at 105°C for 60 minutes, and mix well to obtain the finished product. The viable count of freeze-dried bacteria powder of Bifidobacterium Bbm reached 1.28×10 ¹¹ cfu/g.

Example 2

1. The compound protective agent formula of the present invention: β cyclodextrin 10g, lactose 3g, trehalose 2g, yeast extract 1g, vitamin E 0.06g, water 100mL (adjust the pH value of distilled water to about 7.0 with lye in advance).

2. Preparation:

Accurately weigh and mix the components (except β-cyclodextrin and vitamin E) that make up the composite protective agent; heat to dissolve and cool to room temperature; sterilize at 115°C for 10 minutes and cool to room temperature; add aseptically Filter-sterilized vitamin E and β-cyclodextrin sterilized by dry heat at 105°C for 60 minutes, and mix well to obtain the finished product. The viable count of freeze-dried bacteria powder of Bifidobacterium B1m reached 1.14×10 ¹¹ cfu/g.

Example 3

1. The compound protective agent formula of the present invention: β-cyclodextrin 10g, lactose 4g, trehalose 0.8g, yeast extract 2g, vitamin E 0.1g, water 100mL (adjust the pH value of distilled water to about 7.0 with lye in advance) .

2. Preparation:

Accurately weigh and mix the components (except β-cyclodextrin and vitamin E) that make up the composite protective agent; heat to dissolve and cool to room temperature; sterilize at 115°C for 10 minutes and cool to room temperature; add aseptically Filter-sterilized vitamin E and β-cyclodextrin sterilized by dry heat at 105°C for 60 minutes, and mix well to obtain the finished product. The viable count of freeze-dried bacteria powder of Bifidobacterium Bbm reached 1.20×10 ¹¹ cfu/g.

Example 4

1. Compound protective agent formula: β-cyclodextrin 10g, lactose 2g, trehalose 3g, yeast extract 2g, vitamin E 0.1g, water 100mL (adjust the pH value of distilled water to about 7.0 with lye in advance).

2. Preparation:

Accurately weigh and mix the components (except β-cyclodextrin and vitamin E) that make up the composite protective agent; heat to dissolve and cool to room temperature; sterilize at 115°C for 10 minutes and cool to room temperature; add aseptically Filter-sterilized vitamin E and β-cyclodextrin sterilized by dry heat at 105°C for 60 minutes, and mix well to obtain the finished product. The viable count of freeze-dried bacteria powder of Bifidobacterium Blm reached 1.08×10 ¹¹ cfu/g.

Example 5

1. The compound protective agent formula of the present invention: β-cyclodextrin 10g, lactose 5g, trehalose 2g, yeast extract 0.5g, vitamin E 0.1g, water 100mL (adjust the pH value of distilled water to about 7.0 with lye in advance) .

2. Preparation:

Accurately weigh the components (except β-cyclodextrin and vitamin E) that make up the composite protective agent, and mix to obtain the finished product; heat to dissolve, and cool to room temperature; sterilize at 115°C for 10 minutes, and cool to room temperature; Sterilization operation Add filter-sterilized vitamin E and β-cyclodextrin sterilized by dry heat at 105°C for 60 minutes, and mix well to obtain the finished product. The viable count of freeze-dried bacteria powder of Bifidobacterium Bbm reached 1.11×10 ¹¹ cfu/g.

Claims (6)

1. A compound protective agent for high-activity bifidobacterium dry powder preparation, characterized in that: its formula consists of: β-cyclodextrin 10g, lactose 2-5g, trehalose 0.5-2g, yeast extract 0.5-2g, Vitamin E 0.05-0.1g, distilled water 100mL; the preparation method is as follows: ① Weigh β-cyclodextrin according to the amount, dry heat sterilization at 105°C for 60 minutes, and cool; ②Weigh the vitamin E according to the amount, filter and sterilize; ③Adjust the pH value of the distilled water used to prepare the compound protective agent with lye in advance to 7.0; ④ Add lactose, trehalose and yeast extract in the distilled water with pH adjusted in advance, heat to dissolve and mix well; ⑤ Sterilize at 115°C for 10 minutes and cool to room temperature; ⑥ Add β-cyclodextrin and vitamin E, and mix to obtain the finished product. 2. The composite protective agent according to claim 1, characterized in that its formula consists of: β-cyclodextrin 10g, lactose 5g, trehalose 1g, yeast extract 1g, vitamin E 0.1g, distilled water 100mL. 3. The compound protective agent according to claim 1, characterized in that its formula consists of: β-cyclodextrin 10g, lactose 3g, trehalose 2g, yeast extract 1g, vitamin E 0.06g, distilled water 100mL. 4. The compound protective agent according to claim 1, characterized in that its formula consists of: β-cyclodextrin 10g, lactose 4g, trehalose 0.8g, yeast extract 2g, vitamin E 0.1g, distilled water 100mL. 5. The composite protective agent according to claim 1, characterized in that its formula consists of: β-cyclodextrin 10g, lactose 2g, trehalose 2g, yeast extract 2g, vitamin E 0.1g, distilled water 100mL. 6. The compound protective agent according to claim 1, characterized in that its formula consists of: β-cyclodextrin 10g, lactose 5g, trehalose 2g, yeast extract 0.5g, vitamin E 0.1g, distilled water 100mL.