An acid-resistant Saccharomyces cerevisiae and its application in high-acidity fruit fermented wine
A technology of Saccharomyces cerevisiae and main fermentation, applied in the field of brewing engineering, which can solve the problems of lack of Saccharomyces cerevisiae, low pH during fermentation, insufficient flavor, etc., and achieve the effects of shortened brewing cycle, high acid tolerance and rich aroma
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Embodiment 1
[0059] Example 1: Screening and identification of the starting strain of Saccharomyces cerevisiae ET005-c54
[0060] Specific steps are as follows:
[0061] 1. Screening and identification of wild Saccharomyces cerevisiae
[0062] Saccharomyces cerevisiae was screened from green plums, soil and other samples from different geographical locations in the green plum producing area of Zhao'an County, Fujian Province. The colonies were preliminarily classified and identified by observing the morphology and color, and further tests such as alcohol production, gas production, test-tube fermentation, and acid resistance were carried out. The strains were screened, and finally 10 strains of Saccharomyces cerevisiae with relatively good fermentation performance were selected for further screening and preserved in our laboratory, named YT012, YT018, YT021, ET005, ET016, ET023, SM003, SM007, SM008, and SM012, respectively.
[0063] 2. The 10 strains of Saccharomyces cerevisiae obtained...
Embodiment 2
[0074] Example 2: Obtainment of Saccharomyces cerevisiae ET005-c54
[0075] 1. Mutagenesis of Saccharomyces cerevisiae origin strain ET005 using atmospheric pressure and room temperature plasma mutagenizer
[0076] Specific steps are as follows:
[0077] (1) Pick a ring of the Saccharomyces cerevisiae starting strain ET005 obtained in Example 1 from the plate and inoculate it in a 250 mL conical flask containing 50 mL of liquid seed medium, and let it stand at 30° C. for 10 to 12 hours, so that the bacterial cells are in logarithmic growth. Expect. Take 1 mL of logarithmic growth phase bacterial liquid to measure its OD 600 , according to OD 600 and the general relationship between the number of colonies to estimate the bacterial concentration, at this time the OD 600 After centrifugation at 12000 r / min for 2 min, 1 mL of bacterial solution was collected, washed with sterile saline for 2 to 3 times, and diluted to 10 7 cfu / mL of bacterial suspension.
[0078] (2) Draw 10...
Embodiment 3
[0095] Example 3: Performance verification of Saccharomyces cerevisiae ET005-c54
[0096] 1. Acid resistance test:
[0097] ET005-c54 was placed in the selective medium of pH 1.5, pH 2.0, and pH 2.5, respectively, and incubated at 30 °C for 72 h on a shaker, and its OD was determined by a microplate reader. 600 . like Image 6 As shown, the growth performance of ET005-c54 in the environment of pH 2.5 is good, therefore, it is proved to have better acid resistance.
[0098] The mutant strain ET005-c54 and the starting strain ET005 were inoculated into pH 2.5 selective medium and grown to OD 600 was about 0.8, and the bacterial cells were collected and observed using a cold field emission scanning electron microscope (cFE-SEM). like Figure 7 As shown, at pH 2.5, the mutant strain ET005-c54 was oval and round, the surface was smooth without wrinkles, there were obvious buds and bud scars, most of them were single cells, and there was no adhesion between cells; while in the ...
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