High-throughput screenign method of butanedione high-yield lactobacillus
A screening method and technology for lactic acid bacteria, applied in the field of microbiology, can solve the problems of long time and heavy workload, and achieve the effects of high recovery rate and stability, good correlation, and significant technological progress.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0025] Diacetyl content standard curve preparation:
[0026] (1) 12% concentration of skimmed milk powder is sterilized at 121°C for 7 minutes and then cooled rapidly, which is used to accurately prepare diacetyl standard solutions of different concentrations;
[0027] (2) Get the sample to be tested, add an equal volume of 16% trichloroacetic acid solution, mix well, centrifuge at 3500g for 10min, and get the clarified supernatant for subsequent use;
[0028] (3) Take 200 μL of the supernatant and add it to the 2 wells (A1, B1) of the microtiter plate (100 μL / well), add 5 μL of 1% o-phenylenediamine to A1, do not add B1, shake well and place Place in the dark for 30min, then add 4mol / L HCl solution (A1 plus 20μL, B1 plus 25μL) to stop the reaction, after mixing, use the solution of B1 without o-phenylenediamine as the reference solution, and use the microplate reader at a wavelength of 335nm Measure its OD value. Each sample is parallelized at least 3 times, and the average...
Embodiment 2
[0030] 24-well plate-enzyme standard method to measure the content of diacetyl:
[0031] (1) Take the sample to be tested, add sterile water, 10-fold gradient dilution, shake, take the supernatant and spread it on the MRS agar medium, culture it at 25-30°C for 48-72 hours, and obtain a single colony as the lactic acid bacteria to be screened ;
[0032] (2) Under sterile conditions, take the lactic acid bacteria to be screened, inoculate them in a multi-well plate containing MRS broth, and activate them for 3 generations before use;
[0033] (3) Inoculate 5% of the activated lactic acid bacteria in sterilized skim milk, ferment in a porous plate at 42°C, add an equal volume of 16% trichloroacetic acid after the curd, mix well, and centrifuge at 3500g for 10min. Get clarified supernatant for subsequent use; above-mentioned skim milk composition is as follows: skim milk powder 120g / L, glucose 20g / L, yeast powder 5g / L, peptone 5g / L, solvent is water;
[0034] (4) Take 1600 μL of...
Embodiment 3
[0036] 48-well plate-enzyme standard method to measure the content of diacetyl:
[0037] (1) Take the sample to be tested, add sterile water, 10-fold gradient dilution, shake, take the supernatant and spread it on the MRS agar medium, culture it at 25-30°C for 48-72 hours, and obtain a single colony as the lactic acid bacteria to be screened ;
[0038] (2) Under sterile conditions, take the lactic acid bacteria to be screened, inoculate them in a multi-well plate containing MRS broth, and activate them for 3 generations before use;
[0039](3) Inoculate 5% of the activated lactic acid bacteria in sterilized skim milk, ferment in a porous plate at 42°C, add an equal volume of 16% trichloroacetic acid after the curd, mix well, and centrifuge at 3500g for 10min. Get clarified supernatant for subsequent use; above-mentioned skim milk composition is as follows: skim milk powder 120g / L, glucose 20g / L, yeast powder 5g / L, peptone 5g / L, solvent is water;
[0040] (4) Take 1000 μL of ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More