Method for mutagenizing microbial strains, Clostridium butyricum strains and applications
A technology of Clostridium butyricum and microbial strains is applied in the field of Clostridium butyricum strains and strains of mutagenized microorganisms, which can solve the problems that the frequency of forward mutation cannot be guaranteed to be increased, the mutation has no directionality, etc., and achieves easy operation, Simple and easy operation, low processing cost
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Embodiment 1
[0047] This example is an example of a method for mutagenesis of microorganisms. Specifically, the following steps are included:
[0048] (1) Medium formula
[0049] Growth media, including liquid media and solid media. Liquid medium: glucose 10g / L, yeast powder 8g / L, sodium chloride 2g / L, magnesium sulfate 0.5g / L, pH6.0; solid medium: liquid medium plus agar 15-20g / L, pH6. 0.
[0050] Screening medium: Add 1% calcium carbonate and 0.04% bromocresol violet to the growth medium, pH 7.0.
[0051] (2) Preparation of bacterial suspension: Pick a ring of Clostridium butyricum strain preserved in the laboratory, insert it into the growth medium, activate it, culture it statically at 37°C, and take the seed solution in the logarithmic growth phase for mutagenesis. Centrifuge the seed solution at 5000rpm for 5min, discard the supernatant, wash the precipitate twice with pH7. 7 cfu / mL bacterial suspension.
[0052] (3) Determination of ultraviolet mutagenesis time: take 3mL of th...
Embodiment 2
[0060] This embodiment is an application experiment example of Clostridium butyricum.
[0061] Select 1000 healthy laying hens with uniform body weight, and randomly divide them into 2 groups (control group and Clostridium butyricum group), 500 in each group. The control group was fed with normal chicken feed, and the Clostridium butyricum group was fed with 0.2 g / Kg strain NN-2 on the basis of normal chicken feed. Other than that, chickens in both test groups had free access to feed and water under the same conditions. The test period is 30 days.
[0062] Eggs were collected during the laying period, the egg production rate was counted, and 50 eggs were randomly selected from each group to determine the cholesterol and protein content. It was found that in the Clostridium butyricum group, the egg production rate increased by 2%, the cholesterol in eggs decreased by 33%, and the protein increased by 22.4%, which significantly improved the quality of eggs.
Embodiment 3
[0064] This embodiment is an application experiment example of Clostridium butyricum.
[0065] Add 5% white granulated sugar to raw milk preheated to 63-65°C to prepare, stir to dissolve, homogenize to make it more uniform and stable, heat and sterilize at 95°C, keep warm for 10 minutes and take it out. When the sterilized raw milk is cooled to 42-45°C, add 6% butyric acid bacteria to inoculate, stir and mix well. The inoculated yogurt was fermented and cultured at a constant temperature of 37°C, and the fermentation was terminated after 12 hours. It was taken out and refrigerated at 4°C and cooked for 15 hours to endow the yogurt with a good taste and flavor, and obtain butyric acid bacteria fermented yogurt.
[0066] The prepared butyric acid bacteria fermented yoghurt has delicate texture, soft taste, mellow flavor, smooth surface, stable curd, no stratification, no whey precipitation, unique flavor of yoghurt, and rich nutrition.
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