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Method for constructing novel bacterium belonging to the genus bifidobacterium

一种双歧杆菌属、制作方法的技术，应用在新型双歧杆菌属细菌的制作领域，能够解决低通用性、不能适用、不能发挥存活性改善效果等问题，达到存活性高、利用性高的效果

Active Publication Date: 2012-11-07
YAKULT HONSHA KK
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, these survivability-improving strains have the problem that the survivability-improving effect can only be expected in the environment used in the production method.
That is, in the production of mutant strains of Bifidobacterium bacteria, the method of subculture and storage in an environment where the growth of Bifidobacteria bacteria is difficult to obtain is generally used, and the method of obtaining surviving strains can be expected. There is a certain survivability improvement effect in the environment, but in other environments, the survivability improvement effect cannot be expected
Therefore, the bacteria of the genus Bifidobacterium obtained by the existing method cannot be applied to food and beverages distributed under conditions different from those for producing mutant strains, and have extremely low versatility.
[0007] In addition, although the reason is still unclear, it is known that a strain with improved viability obtained by subculture storage under conditions where environmental factors deteriorate (for example, acidic region pH) is suitable for strains at neutral region pH where more stable conditions , and the viability improvement effect cannot be exhibited, which is one of the reasons why it is not possible to obtain a highly versatile strain applicable to various food and beverages

Method used

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  • Method for constructing novel bacterium belonging to the genus bifidobacterium
  • Method for constructing novel bacterium belonging to the genus bifidobacterium
  • Method for constructing novel bacterium belonging to the genus bifidobacterium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1）

[0076] (Example 1) Breeding improvement of Bifidobacterium breve YIT 10001 and YIT4125 obtained by alternate subculture

[0077] Bifidobacterium breve strain YIT 10001 (FERM BP-8205) and Bifidobacterium breve strain YIT4125 (FERM BP-7813) were used as parent strains for alternate passage and enrichment.

[0078] (1) Sterilize 20.7% whole milk powder medium at 135°C for 3.5 seconds, and then inoculate 0.5% seed starter of Bifidobacterium breve YIT 10001 strain or YIT 4125 strain, 1% Bifidobacterium breve The inoculum of bifidobacteria was mixed and cultured at 33°C until the pH was 5.3, and the bacterial solution A1 (400mL) was prepared.

[0079] (2) Blend syrup A into bacterial solution A1 so that the final concentration of palatinose is 10%, and prepare dairy product A1 (630 mL). Divide dairy product A1 (200mL) into 300mL flasks, stir (90rpm) at 5°C for 1 week under aerobic conditions with cotton plugs placed, store (hereinafter referred to as aerobic stirring storage), and ...

Embodiment 2）

[0085] (Example 2) Viability confirmation test

[0086] Using the parent strain and the isolated strain, prepare dairy products A and B respectively according to the method in Example 1, store them under aerobic conditions at 5°C with stirring for 1 week, and then store them statically at 10°C under anaerobic conditions for 2 weeks . Bifidobacterium breve YIT 12272 strain (from YIT 4125, isolated from dairy product A8) with good survival in both dairy products A and B was selected.

[0087] Table 3 shows the results of viability in dairy products A and B of YIT 12272 strain and control bacteria (YIT 10001 strain, YIT 4125 strain). Regardless of whether YIT 12272 strains were stored in either dairy products A or B, 3×10 7 Viable bacteria above CFU / mL. On the other hand, any one of the dairy products A and B made with the control bacteria left 3×10 7 Live bacteria above CFU / mL, but the products of the two cannot show good viability.

[0088] In addition, the physical proper...

Embodiment 3）

[0098] (Example 3) Confirmation test of artificial gastric juice tolerance

[0099] Bifidobacterium breve YIT 12272 strain and control bacteria (YIT 10001 strain, YIT 4125 strain) were anaerobically cultured in milk medium at 37°C overnight. Add 0.5 mL of this bacteria solution to 10 mL of artificial gastric juice warmed at 37°C for 30 minutes, mix quickly, and then keep warm at 37°C. After incubation (artificial gastric juice treatment) for 0 minutes and 120 minutes, 1 mL was aliquoted, and after appropriate dilution, the number of viable bacteria was measured in TOS propionate agar medium (Yakult Pharmaceutical Industry, 37°C, anaerobic culture).

[0100] Among them, the artificial gastric juice was prepared as follows. That is, dissolve in ion-exchanged water so that the final concentration is peptone (Becton, Dickinson and Co.): 5 g / L, gastric mucosin (Wako Pure Chemical Industries): 1.5 g / L, sodium chloride: 5 g / L, Sodium bicarbonate: 3g / L, sodium dihydrogen phosphate: ...

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Abstract

Provided are a method for constructing a bacterium belonging to the genus Bifidobacterium which shows excellent survival even under conditions with different environmental factors, the novel bacterium belonging to the genus Bifidobacterium constructed by said method, and a method for detecting said bacterium. By alternately conducting subcultures and preservations in systems under such conditions as differing from each other in environmental factors, a bacterium belonging to the genus Bifidobacterium, which shows excellent survival under all of the conditions employed in the alternate subcultures and preservations, can be constructed.

Description

technical field [0001] The invention relates to a preparation method of a novel Bifidobacterium bacterium, a novel Bifidobacterium bacterium obtained according to the preparation method and a detection method of the bacterium. Background technique [0002] Bacteria belonging to the genus Bifidobacterium are the main bacteria in the bacterial colonies in the human intestine, and are known to have beneficial effects on human health such as bowel-regulating effects such as improving constipation and diarrhea, suppressing the increase in serum cholesterol, and activating immunity. Therefore, various commercial products are sold in the form of various fermented foods and beverages, live bacterial preparations, etc., and fermented milk foods and beverages in particular have excellent palatability and are therefore suitable for continuous ingestion of bacteria belonging to the genus Bifidobacterium. [0003] Bacteria belonging to the genus Bifidobacterium are obligate anaerobic bac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23C9/127A23L1/30A23L2/02C12N15/09C12Q1/04C12Q1/06
CPCC12Q1/689A23Y2300/25A23Y2300/00A23L1/3014C12Q1/04A23Y2240/41A23Y2300/29A23C9/1234C12N1/36A23Y2220/03A23C9/1236A23L2/52C12R1/01C12Q1/06A23L33/135C12R2001/01C12N1/205A23V2400/231A23V2400/517A23V2400/519A23V2400/113A23V2400/51C12N1/20
Inventor 左古知行三浦美加岛川康久宫崎幸司藤本淳治渡边幸一
Owner YAKULT HONSHA KK
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