A kind of acid-reducing yeast strain and its application
A technology of yeast and strains, applied in the field of microorganisms, can solve the problems of poor degradation ability and poor application of acid-reducing treatment, and achieve the effects of low cost, improved flavor quality, and simple treatment methods
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Embodiment 1
[0021] Example 1 Screening and isolation of strains
[0022] Slightly rotten navel orange and papaya were used as experimental raw materials, chopped with skin and added with 9 times the volume of sterile water, and cultured for 50 min in a constant temperature shaker at a speed of 150 r / min and a temperature of 28 °C.
[0023] Add 10% (v:v) cultured sample solution to liquid Sabouraud medium, and cultivate for 24-48h at 150r / min and 28°C, and observe until the fermentation broth is turbid.
[0024] The pH of solid Sabouraud medium was adjusted to 4 with 20% citric acid stock solution, and 10 was adjusted by coating method. -4 , 10 -5 , 10 -6 , 10 -7 The dilution gradient of the fermentation broth was evenly spread on the plate, and incubated at a constant temperature of 28°C for 48-96h until colonies appeared.
[0025] The isolated colonies were numbered and expanded, and a small amount of colonies were picked and inoculated into the liquid Sabouraud medium, activated and...
Embodiment 2
[0027] Embodiment 2 Bacteria acid-reducing effect test
[0028] First of all, the overall acid-reducing effect of the six strains obtained was evaluated. The specific methods are as follows:
[0029] Preparation of raspberry juice: mix fresh raspberry fruit and water according to the quality of 1:1, use a juicer (300W) to squeeze juice at a constant speed for 1min, and then use a homogenizer (10000rpm) to homogenize for 5min to obtain a uniform whole raspberry juice. juice. It was then sealed and pasteurized at 80°C for 15 minutes.
[0030] The above 6 kinds of non-Saccharomyces cerevisiae were activated and expanded, added to the above fruit juice at a volume fraction of 5%, and dynamically cultivated at a constant temperature of 28 ° C for 72 hours, during which pH was measured every 12 hours, and the results were as follows figure 1 shown.
[0031] from figure 1 It can be seen that the pH of the juice inoculated with L-2 and L-3 strains began to rise from 36h after inoc...
Embodiment 3
[0040] The identification of embodiment 3 strains
[0041] 1. DNA extraction
[0042] (1) Take 1 mL of the bacterial solution to be tested and centrifuge at 10,000 r / min for 2 minutes, take the precipitate and add 400 μL of DNA extraction buffer, then put it in boiling water and boil for 10 minutes, then take out the ice bath for 10 minutes, and then take the water bath at 65°C for 10 minutes.
[0043] (2) 250 μL of 6mol / L NaCl solution and equal volume of chloroform: isoamyl alcohol (24:1) were added thereto, mixed for 10 min, and centrifuged at 13000 r / min for 5 min in a centrifuge.
[0044] (3) Take the centrifuged supernatant, add twice the volume of anhydrous ethanol, invert and mix, let stand for 5 minutes, and centrifuge in a centrifuge at 13000 r / min for 5 minutes.
[0045] (4) Discard the supernatant, wash the precipitate once with 500 μL of 70% ethanol, and centrifuge in a centrifuge at 13000 r / min for 3 min.
[0046] (5) Carefully aspirate the supernatant, and after...
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