Method for increasing total acid content in gluconobacter oxydans fermented vinegar
A technology of Gluconobacter oxidans and total acid content, which is applied in the field of microbial application and vinegar brewing, can solve the problems of decreased acid production and death of acetic acid bacteria, and achieves the effects of improving acid resistance, reducing damage and improving integrity
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Embodiment 1
[0028] Embodiment 1: Ultrasonic treatment of Gluconobacter oxydans W-1 high-alcohol fermented vinegar
[0029] (1) Preparation of seed solution
[0030] Inoculate the strains of Gluconobacter oxydans W-1 and Huyao 1.01 in the liquid seed medium, the composition of which is: glucose, yeast extract, ethanol and distilled water; pH 6.0, 30°C, 180rpm for 24h, Prepare the seed solution.
[0031] (2) Ultrasonic fermentation
[0032] Inoculate the seed liquid of Gluconobacter oxydans W-1 and Shanghaiyao 1.01 strains into the liquid fermentation medium at 5% (v / v), pH 6.0, 30°C, 180rpm for 1 day and then ultrasonic treatment, the power is 350W, and the time is 25min; the fermentation time is 8 days.
[0033] Specifically, a specific composition of the fermentation medium is: glucose 20g / L, yeast extract powder 10g / L, ethanol 80mL / L, pH 6.0.
[0034] Depend on figure 1 It can be seen that the biomass of Gluconobacter oxydans W-1 treated with ultrasonic wave has no significant chan...
Embodiment 2
[0036] Example 2: Ultrasonic treatment of changes in cell morphology and microstructure of Gluconobacter oxydans W-1
[0037] (1) Treat the bacteria and observe with electron microscope
[0038] Scanning and transmission electron microscopy (SEM and TEM) were used to observe the morphological and ultrastructural changes of bacteria after sonication. Each 1 mL sample was centrifuged at 6000 rpm for 10 minutes. Cell pellets of samples were fixed with 2.5% (v / v) pre-cooled glutaraldehyde at 4°C for 2 hours. Next, samples are dehydrated, embedded and prepared for ultrathin sections. The cross section of each sample was then observed under a transmission electron microscope (JEM-1200EM).
[0039] (2) SEM image
[0040] Depend on image 3 It can be seen that the supersonic treatment of Gluconobacter oxidans W-1 is a bacillus, which is more slender and smoother than before treatment. However, the shape of Gluconobacter oxydans W-1 without sonication was close to ellipse, the ce...
Embodiment 3
[0043] Example 3: Changes in the activity of alcohol dehydrogenase and acetaldehyde dehydrogenase of Gluconobacter oxydans W-1 by ultrasonic treatment
[0044] (1) Preparation of crude enzyme solution
[0045] Take the fermentation broth of Gluconobacter oxidans W-1 at 4°C, centrifuge at 8000g for 10min, and discard the supernatant. Then resuspend the cells with 20 mL of pre-cooled KPB (50 mmol / L, pH 6.0) buffer, centrifuge at 8000 g for 10 min, discard the supernatant, repeat the above operation, and wash the cell pellet 3 times. Then add 20mL of KPB buffer to resuspend the bacteria, and use SCIENTZ-IID ultrasonic cell pulverizer to perform ultrasonic crushing. The ultrasonic crushing conditions are: crushing time: 5min; crushing on / off=3 / 3s; crushing power: 40%. After crushing, the crushed solution was centrifuged at 4°C, 8000r / min for 5min, and the obtained supernatant was the crude enzyme solution.
[0046] (2) Determination of ADH and ALDH enzyme activity
[0047] Take...
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