Lactic acid bacterium having immunoregulatory activity derived from moromi for wine fermentation
A technology of immune regulation and lactic acid bacteria, applied in the preparation of wine, alcoholic beverages, medical raw materials derived from bacteria, etc., can solve the problems that the effect has not been studied
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Embodiment 1
[0048] 〔Using lactic acid bacteria strain〕
[0049] In order to select strains with strong Th1 immune activation ability, 104 strains of lactic acid bacteria isolated from wine brewing mash were used in the comparative experiment of interleukin 12 (IL-12) induction ability.
[0050] [Induction of IL-12 stimulated in vitro]
[0051] For BALB / c mice (7 weeks old, male), 4.05% sodium thioglycolate was administered intraperitoneally, and intraperitoneal macrophages were recovered using PBS 4 days later, and adjusted to 2 ×10 6 After cells / mL, add 0.5 mL per well in a 24-well (well) plate. Heat-dead bacteria (1 μg / mL) of each strain were added to each well, and the IL-12 concentration in the culture supernatant after 24 hours of culture was measured. No dead bacteria were added to the control. IL-12 (p70) was measured because the active form of IL-12 was p70 in which subunits of p35 and p40 combined. In addition, an OptEIA mouse IL-12 assay kit (manufactured by BDPharmingen) w...
Embodiment 2
[0060] 〔Using lactic acid bacteria strain〕
[0061] The 12 strains of lactic acid bacteria selected in Example 1 were used in the comparison experiment of stimulating IL-12 inducing ability in vivo.
[0062] [IL-12 induction by stimulation in vivo]
[0063] To BALB / c mice (7 weeks old, male), the heat-killed bacteria (500μg / 0.2mL / mouse) suspension (solvent: physiological saline) of each of the above strains was intraperitoneally administered, and the cervical vertebrae were dislocated 6 hours later. Afterwards, blood was collected from the heart. The same amount of physiological saline was administered to the control mice. After blood collection, serum was collected by centrifugation, and the IL-12 concentration in the serum was measured using an OptEIA mouse IL-12 assay kit (manufactured by BD Pharmingen).
[0064] The result is as figure 1 shown. Serum IL-12 concentrations were particularly high in the A and L strains. Therefore, they were named SAM2446 strain and SAM2...
Embodiment 3
[0065] Example 3 Identification of SAM2446 and SAM2447 strains
[0066] The research results of the mycological properties of the SAM2446 strain are shown in Table 1, and the research results of the SAM2447 strains are shown in Table 2.
[0067] Based on these findings, it was deduced that the SAM2446 strain is Lactobacillus plantarum and the SAM2447 strain is Lactobacillus brevis. It is known that there are two kinds of Lactobacillus plantarum with sorbitol assimilation ability and those without sorbitol assimilation ability. As shown in Table 1, SAM2446 strain is a lactic acid bacteria species with sorbitol assimilation ability. And it is known that there are two types of Lactobacillus brevis with and without D-xylose assimilation. As shown in Table 2, SAM2447 strain is a lactic acid bacteria species without D-xylose assimilation.
[0068] After extracting DNA from SAM2446 and SAM2447 strains, the 16S rRNA gene region was deciphered. Because the 16SrRNA gene sequence (sequ...
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