Bacillus natto capable of producing MK-7 and application of bacillus natto
A technology of Bacillus natto and MK-7, applied in the field of bioengineering, can solve the problems of increasing atherosclerosis and blood pressure, and achieve the effect of improving osteoporosis.
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Embodiment 1
[0043] Primary screening of Bacillus natto
[0044] 1 Materials and methods:
[0045] 1.1 Experimental strains, Bacillus natto BLCC1-0048, Bacillus natto BLCC1-0053, Bacillus natto BLCC1-0054;
[0046] 1.2 Fermentation medium and culture conditions:
[0047] Two media formulations:
[0048] Seed medium: glucose 0.2%, peptone 1%, beef extract 0.5%, NaCl 0.5%, pH 7.2-7.4;
[0049] Fermentation medium: yeast extract 5%, glycerol 5%, peptone 10%, K 2 HPO 4 0.06%;
[0050] Culture conditions: culture on a shaker at 37°C for 24h, with a rotation speed of 120rpm, and then culture at 45°C for 5 days.
[0051] 1.3 After the fermentation is finished, the fermentation broth and bacteria are separated by centrifugation, and the bacteria are freeze-dried.
[0052] 1.4 Determination of MK-7 content in fermentation cells and fermentation broth:
[0053] Weigh an appropriate amount of freeze-dried bacteria (about 25mg), extract it with a mixed solution of chloroform: n-hexane = 2:1 (...
Embodiment 2
[0063] Determination of MK-7 content in Bacillus natto fermented for 6 days under different medium conditions:
[0064] 1 Materials and methods:
[0065] 1.1 Experimental strain Bacillus natto strain BLCC 10053;
[0066] 1.2 Fermentation medium and culture conditions:
[0067] Three basal media formulations:
[0068] N1: Glycerol 3%, Peptone 10%, NaCl 0.5%, K 2 HPO 4 0.02%, pH 7.0-7.2
[0069] N2: glucose 0.2%, yeast extract 0.5%, peptone 1.0%, sodium chloride 0.5%, pH 7.0-7.2
[0070] N3: yeast extract 2.5%, glycerol 2.5%, peptone 9.5%, K 2 HPO 4 0.03%, pH 7.0-7.2
[0071] Training conditions:
[0072] The specific experimental scheme is shown in Table 2. The activated Bacillus natto was inserted into the seed medium, cultured on a shaking table at 42°C for 16 hours, transferred to the fermentation medium with an inoculum size of 5%, and cultured on a shaking table at 37°C for 1 day. Transfer to a 45°C incubator for static culture for 5 days.
[0073] 1.3 After t...
Embodiment 3
[0083] Content of Bacillus natto MK-7 under different fermentation time conditions:
[0084] 1 Materials and methods:
[0085] 1.1 Experimental strain Bacillus natto strain BLCC 10053;
[0086] 1.2 Fermentation medium and culture conditions;
[0087] Seed medium N2: glucose 0.2%, peptone 1.0%, sodium chloride 0.5%, yeast extract 0.5%, pH 7.0-7.2; fermentation medium N1: peptone 10%, glycerol 3%, NaCl 0.5%, K 2 HPO 4 0.02%, pH 7.0-7.2.
[0088] The activated Bacillus natto was inserted into the seed medium, cultured on a shaker at 42°C for 16 hours, transferred to the fermentation medium at an inoculum size of 5%, cultured on a shaker at 37°C for 1 day, and then transferred to an incubator at 45°C for standing Culture 3d, 4d, 5d, 6d, 7d.
[0089] 1.3 After the fermentation is finished, the fermentation product and bacteria are separated by centrifugation, and the bacteria are freeze-dried.
[0090] 1.4 Determination of MK-7 content in fermented bacteria and fermented liqu...
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