Preparation method and application of anti-aging and anti-wrinkle yeast extract
A technology of yeast extract and Saccharomyces cerevisiae, which is applied in the field of preparation of yeast extract, can solve the problems of high cost, complicated preparation process, high quality standard, etc., and achieve the effect of precise classification and positioning, broad market prospect and huge economic value
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Embodiment 1
[0131] Embodiment 1 mutagenizes industrial Saccharomyces cerevisiae strains by ARTP mutagenesis method
[0132] 1. Activation and fermentation of industrial Saccharomyces cerevisiae
[0133] Starting strain: Industrial Saccharomyces cerevisiae (Saccharomyces cerevisiae, ATCC 9080);
[0134] The Saccharomyces cerevisiae strain was inserted into the fermentation medium, and cultured with shaking at 30°C and 150r / min until the bacteria grew to the logarithmic phase.
[0135] 2. Using the ARTP mutagenesis method to mutate Saccharomyces cerevisiae strains
[0136] Take 1 mL of the cultured bacterial solution to the mid-logarithmic phase, put it in a 1.5 mL EP tube, centrifuge at 8000 r / min at 4 °C for 5 min, and discard the supernatant;
[0137] After washing 3 times with normal saline containing glycerol (mass fraction 5%), draw 10 μL of the suspension and place it on a cell counting plate, count the number of bacteria with a microscope, and dilute the bacteria to a concentratio...
Embodiment 2
[0166] Example 2 Purification of Yeast Extract by Molecular Sieve and Heparin Affinity Chromatography
[0167] 1. Yeast cell treatment
[0168] Use resuspension (20mM Tris-HCl, 0.1M NaCl, pH 7.0) to resuspend JCY-P-998 bacteria according to the wet weight of the bacteria (g): resuspension (mL) = 1:15 (10000rpm, 3min ), tissue mashing homogenizer (high pressure homogenization 800bar) 2 times of homogenization;
[0169] Insert the probe into the homogenate, about 3cm away from the bottom of the container, set the power at 10%, sonicate for 3s each time, with an interval of 5s, sonicate for 20min, and sonicate for 3 times. This process is carried out in an ice bath;
[0170] After the bacteria are broken, use a large-scale low-temperature centrifuge at 4°C, centrifuge at 17,000g for 30min, and collect the supernatant;
[0171] Then, the supernatant was filtered with a 0.45 μm filter membrane and loaded onto molecular sieves.
[0172] 2. SuperdexTM30prep grade molecular sieve p...
Embodiment 3
[0181] Embodiment 3 yeast extract detection method
[0182] 1. Determination of bacterial biomass
[0183] Take 50mL of Saccharomyces cerevisiae JCY-P-998 fermentation broth, put it in a 100mL centrifuge tube, centrifuge at 4°C, 8000r / min for 15min, and discard the supernatant;
[0184] Washing with distilled water-centrifugation was repeated 3 times, and the supernatant was discarded; the bacterial cell pellet was dried in a desiccator at 60°C to constant weight, cooled in a desiccator to room temperature and weighed.
[0185] 2. Detection of genetic stability of yeast strain JCY-P-998
[0186] The screened Saccharomyces cerevisiae strain JCY-P-998 was continuously passaged for 8 generations, and the strains of different generations were used for fermentation and culture, and the biomass and polypeptide content of the fermentation products were detected;
[0187] 3. Yeast extract YL-004 polypeptide content determination
[0188] Accurately measure 1mL of the YL-004 solutio...
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