Clostridium tyrobutyricum and application thereof
A clostridium tyrobutyricum, reaction technology, applied in the direction of bacteria, isolated microorganisms, microorganisms, etc., to reduce the risk of allergies and reduce the effect of morbidity
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Embodiment 1
[0022] Example 1 Screening method for Clostridium tyrobutyricum
[0023] Take 10 g of feces samples from Xinjiang brown cattle, mix them evenly, add them to 90 mL of PBS buffer, put them in a water bath at 100 °C for 10 min to kill non-spore bacteria, and transfer them to RCM medium for anaerobic culture at 37 °C for 24 h; In the logarithmic phase of the growth process, carbon sources such as starch and lignocellulose are added in batches, and Clostridium tyrobutyricum can decompose and use various carbon sources for enrichment culture, which can improve its own vitality and reduce E. coli in feces After culturing for 24 hours, transfer the above culture solution into TSN medium, and culture it anaerobically at 37°C for 24 hours; The resulting samples were diluted to 10 -2 、10 -4 、10 -6 , spread 100 μL on a whey protein isolate plate, and incubate at 37°C for 48 h; select colonies with hydrolysis circles, transfer them to a new whey protein isolate plate, and incubate at 37...
Embodiment 2
[0025] Example 2 Degradation effect of Clostridium tyrobutyricum suspension on α-lactalbumin and β-lactoglobulin
[0026] Drawing of the standard curve: Accurately weigh 8 mg of β-lactoglobulin standard and α-lactalbumin standard, add 2 mL of PBS to dissolve, prepare a standard stock solution of 4 mg / mL, draw an appropriate amount of stock solution, add PBS to dilute, Diluted to 4, 2, 1, 0.5, 0.25, 0.125, 0.0625, 0.03125 mg / mL concentration gradient standard solution for determination. Chromatographic column: Bio-C8 (250mm × 4.6mm, 5μm); mobile phase: 0.1% trifluoroacetic acid (TFA) water (A), 0.1% trifluoroacetic acid (TFA) acetonitrile (B), gradient elution conditions: 0-17.5 min, B 70%-50%; 17.5-20 min, B 50%-70%. Column temperature: 35°C; flow rate: 1.0 mL / min; injection volume: 10 μL; detection wavelength: 280 nm. Draw a standard curve based on standard concentration and peak area.
[0027] Clostridium tyrobutyricum ( Clostridium tyrobutyricum ) Z816 was inoculated i...
Embodiment 3
[0031] Example 3 Degradation effect of Clostridium tyrobutyricum cell disruption liquid on α-lactalbumin and β-lactoglobulin
[0032] Drawing of the standard curve: Accurately weigh 8 mg of β-lactoglobulin standard and α-lactalbumin standard, add 2 mL of PBS to dissolve, prepare a standard stock solution of 4 mg / mL, draw an appropriate amount of stock solution, add PBS to dilute, Diluted to 4, 2, 1, 0.5, 0.25, 0.125, 0.0625, 0.03125 mg / mL concentration gradient standard solution for determination. Chromatographic column: Bio-C8 (250 mm × 4.6 mm, 5 μm); mobile phase: 0.1% trifluoroacetic acid (TFA) water (A), 0.1% trifluoroacetic acid (TFA) acetonitrile (B), gradient elution Conditions: 0-17.5 min, B 70%-50%; 17.5-20 min, B 50%-70%. Column temperature: 35°C; flow rate: 1.0mL / min; injection volume: 10 μL; detection wavelength: 280 nm. Draw a standard curve based on standard concentration and peak area.
[0033] Clostridium tyrobutyricum ( Clostridium tyrobutyricum ) Z816 wa...
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