Application of bacillus coagulans BC99 in preparation of selenium-enriched medicine or food
A Bacillus coagulans, selenium-enriched technology, applied in the field of microorganisms, can solve the problems of selenomethionine unsafety, affecting protein activity, etc., and achieve the effects of strong gastric acid resistance, bile salt resistance, strong selenium-enrichment ability, and wide application prospects
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Embodiment 1
[0025] Example 1: Screening of selenium-resistant Bacillus coagulans BC99
[0026] The 30 high-efficiency strains are derived from the Weikang probiotic strain resource bank and obtained by the screening method in the published patent CN202110285880.1A.
[0027] Strain activation: 30 strains of Bacillus coagulans were streaked on the plate of beef extract peptone solid medium, and cultivated in a constant temperature incubator at 42°C for 48 hours to obtain a plate with a single colony; pick a single colony and insert it into the above seed medium medium, 42°C, 250rpm shaker for 24h to obtain seed liquid;
[0028] Screening of selenium-resistant strains: the above-mentioned seed solution was inoculated into seed medium containing different concentrations of sodium selenite (0, 10, 20, 30, 40 μg / mL) according to the inoculum amount of 2% (V / V), and cultured at 42°C After 24 hours, the color changes and growth of different Bacillus coagulans were observed. The Bacillus coagula...
Embodiment 2
[0032] Example 2: Determination of selenium-enriched content of Bacillus coagulans BC99
[0033] The selenium content determination method adopts the fluorescence spectrophotometry in the national standard "GB5009.93-2017 Determination of selenium in food".
[0034] Hydrochloric acid solution (1%): Measure 5 mL of hydrochloric acid, dilute to 500 mL with water, and mix well.
[0035] Hydrochloric acid solution (6 mol / L): Measure 50 mL of hydrochloric acid, slowly add it to 40 mL of water, and after cooling, dilute to 100 mL with water.
[0036] EDTA-hydroxylamine chloride mixed solution: each 250 mL of mixed solution contains 2.5 g of disodium EDTA and 6.25 g of hydroxylamine chloride, prepared with distilled water.
[0037] DAN reagent (1g / L): This reagent is prepared in a dark room. Weigh 0.2 g of DAN into a conical flask with a cap, add 200 mL of 1% hydrochloric acid solution, and shake for about 15 minutes to dissolve it completely. Add about 40mL of cyclohexane, and co...
Embodiment 3
[0045] Example 3: Determination of Antioxidant Ability of BC99
[0046] (1) Preparation of lactic acid bacteria intact cell IC
[0047] The above-mentioned activated BC99 and selenium-enriched BC99 bacteria were collected by centrifugation at 1000 r / min, washed with PBS for 2 times and resuspended, and the number of bacteria was adjusted to 10. 9 The concentration is the lactic acid bacteria complete cell (IC) suspension;
[0048] (2) DPPH scavenging experiment
[0049] The 0.8mL sample (the number of viable bacteria was controlled to be 10 9 concentration) was added to 1 mL of 0.2 mmol / L freshly prepared DPPH-anhydrous ethanol solution, mixed well, reacted in the dark for 30 min at room temperature, centrifuged at 10,000 r / min for 2 min, and the supernatant was taken to measure the absorbance at a wavelength of 517 nm, Denoted as As. The blank group replaced the DPPH solution with an equal volume of anhydrous ethanol to measure the absorbance, denoted as Ab; the control g...
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