Polysaccharide produced by microorganism belonging to genus bifidobacterium
一种微生物、多糖的技术,应用在细菌、药物组合、食物科学等方向,能够解决功能未明确等问题
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Embodiment 1
[0094] (Example 1: isolation of microorganisms)
[0095] (preparation of medium)
[0096] As a medium for isolating bifidobacteria, BL agar medium (Nissui Pharmaceutical Co., Ltd.) and BS agar medium were used. The BL agar medium was sterilized in an autoclave at 115° C. for 20 minutes, added 5% horse blood under aseptic conditions, and dispensed into petri dishes for use as plates. BS agar medium was prepared as follows. First, 30 g of sodium propionate, 100 mg of paromomycin sulfate, 400 mg of neomycin sulfate, and 6 g of lithium chloride were dissolved in purified water to make the total amount 100 ml, and an additive solution for BS agar medium was prepared. 50 ml of this supplemented BS agar medium was added to 1000 ml of BL agar medium to obtain a BS agar medium. This BS agar medium was dispensed onto a petri dish and used as a plate.
[0097] (Screening of microorganisms)
[0098] Human feces were dissolved in sterile water, inoculated on the above-mentioned BL aga...
Embodiment 2
[0099] (Example 2: Structural Analysis of Polysaccharides)
[0100] (preparation of medium)
[0101] Pancreatin preparation (Amano Enzyme Co., Ltd.) and silicon were added to a 9% by mass skim milk solution so that the final concentrations were 0.36% by mass and 0.01% by mass, respectively. Next, the pH of the solution was adjusted to 8 with 10N NaOH, and reacted at 55° C. for 4 hours to obtain enzymatic skim milk. Cultivator (Yaizu Suisan Chemical Industry Co., Ltd.), lactose, and sodium ascorbate were added thereto so that the final concentrations were 3.0% by mass, 2.5% by mass, and 0.2% by mass, respectively, and sterilized with an autoclave at 121° C. for 15 minutes. Used as a liquid medium.
[0102] (purification of polysaccharides)
[0103] The Bifidobacterium longum JBL05 strain (NITE BP-82) pre-cultivated with the liquid medium prepared above was inoculated into the same liquid medium at a concentration of 1% (v / v), and carried out at 37°C for 40 hours. Static ana...
Embodiment 3
[0118] (Example 3: cultivation and production of polysaccharides)
[0119] Sodium ascorbate was added to the enzymatically digested skim milk prepared in Example 2 so that the final concentration was 0.2% by mass, and it was sterilized with an autoclave at 121° C. for 15 minutes to prepare a liquid medium. The culture solution of Bifidobacterium longum JBL05 strain (NITE BP-82) that is pre-cultivated with the same liquid medium is inoculated in the liquid medium prepared above, and the concentration is 1% (v / v), stirred at a low speed, The pH was controlled at 6.0, and anaerobic culture was carried out at 37°C for 40 hours. The pH was controlled using 10N NaOH. The case where static anaerobic culture was performed was set as a comparative example.
[0120] Fig. 1 shows the relationship between the culture time and the viscosity of the culture solution. Compared with the case of static anaerobic culture, when the pH was adjusted and the culture was performed, the viscosity i...
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