Bacillus acidovorax capable of producing cream fruity flavor and application of bacillus acidovorax
A technology of Bacillus acidicus and creamy fruity aroma, applied in the field of microbiology, can solve the problems of undiscovered patent publications, etc., and achieve the effect of rich flavor, lubricated and mellow taste, and soft taste
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Embodiment 1
[0043] Embodiment 1: the separation of acid dwelling bacillus
[0044] The vinegar samples used come from rural rice vinegar in Guangdong Province. The samples were taken from the fermentation tanks fermented for 15 days and 30 days respectively. The five-point method was used to sample the samples. After being thoroughly mixed, they were placed in sterile EP tubes and stored in a -80°C refrigerator. Weigh 5.0mL vinegar sample under aseptic conditions, mix it with 45mL sterile saline, and shake it in a shaker for 2h to disperse the microbial cells, and make 10 -1 Dilution of the dilution, and serial dilutions. Take 0.1mL respectively to dilute to 10 -5 、10 -6 and 10 -7 Inoculate the dilution of the above-mentioned solution into YPD solid medium, and after coating, place the plates in a common incubator at 30°C for 48 hours, observe and pick individual colonies, and perform repeated streaking and purification to obtain pure strains. Preserve in glycerol tubes, mix the bacte...
Embodiment 2
[0046] Embodiment 2: Molecular biology identification of acid dwelling bacillus
[0047] Cultivate the activated strain to the logarithmic phase, take 1mL of the bacterial liquid, centrifuge at 8000r / min to collect the bacterial cells, add 100μL of sterile water and mix well, so that the cell concentration is about 10 5 individual / μL. Take 10 μL and add to 90 μL sterile water to dilute 10 times to prepare 10 5 、10 4 、10 3 、10 2 and 10 cells / μL concentration of bacterial solution, inoculate 3 μL of each dilution on the plate, and observe the growth of the colony. Select the strains that can grow on the plate for identification.
[0048] The DNA extraction kit step is to extract the genome of the tested strain, perform PCR amplification with bacterial 16S rDNA sequence universal primers (27F and 1492R), send the PCR amplification product to Jinweizhi Biotechnology Company for sequencing, and post the spliced sequence results in GenBank A BLAST homology search was perform...
Embodiment 3
[0049] Example 3: Determination of the highest ethanol tolerance of Bacillus acidicus
[0050] Select the better growing Bacillus acididioides to further study its growth in ethanol environment. The activated bacterial strain was inoculated into the liquid medium containing 0%, 1%, 2%, 4%, 5%, 6% and 7% ethanol (v / v) with 3% (v / v) inoculum, and the bacterial strain was measured. The absorbance of the solution was used to observe the growth ability.
[0051] The results of the growth curve determination of bacterial strains under 7 kinds of ethanol contents are as follows: image 3 , the growth curve of the strain reflects the growth ability of the strain. The growth of the bacteria in the medium with 0% ethanol content is the best. In the medium with 1%, 2% and 4% ethanol content, the growth rate of the strain is slightly slow, but it has no difference with the strain cultured in the medium with 0% ethanol content. Big difference. In the first 30 hours, although the growth...
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