Screening and application of yeast CGMCC 4747 for high production of ethanol and low production of fusel oil in production of Chinese Maotai-flavor liquor
A technology of fusel oil and Saccharomyces cerevisiae, applied in the field of bioengineering, can solve problems such as various influencing factors, production risks, and complex processes, and achieve the effects of reducing production costs, improving wine quality, and optimizing production processes
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Embodiment 1
[0031] Example 1: Screening of Yeast Strains with High Ethanol Production and Low Fusel Oil Production
[0032] Dissolve 0.5g of fermented grains in 15mL sterile saline shaker and shake for 30 minutes, then dilute, spread on a TTC primary screening plate, select red colonies, and then transfer to shake flask culture for secondary screening. The content of ethanol and fusel oil was determined by specific gravity method and HS-SPME-GC-MS, and the strains with high ethanol concentration and low fusel oil concentration were screened.
[0033]TTC primary screening culture: TTC upper layer medium (g / L): glucose 5, agar 15, sterilized at 121°C for 20 minutes, when cooled to about 60°C, add TTC solution (triphenyltetrazolium hydrochloride, final concentration 0.5), immediately pour the bottom plate. TTC lower layer: YPD agar. Cultivate at 30°C for 24 hours. After visible colonies grow, pour about 12mL of TTC upper layer medium to cover the original colonies, and then incubate at 30°...
Embodiment 2
[0035] Example 2: Yeast with high ethanol production and low fusel oil production and its molecular biological identification
[0036] Molecular biological identification was carried out on the obtained yeasts with high ethanol production and low fusel oil production, and the 26srDNA fragments of the strains were amplified with yeast-specific classification and identification primers, and detected by gel electrophoresis. Sequencing comparison was then carried out to determine the species of the screened yeast. The strains were deposited in the General Microbiology Center of the China Committee for the Collection of Microbial Cultures. See Table 1 for the yeast species and preservation numbers.
[0037] Table 1 Yeast with high ethanol production and low fusel oil production
[0038] serial number Chinese name English name Deposit number 1 Pichia anomalies Pichia anomala CGMCC No.4740 2 Schizosaccharomyces pombe Schizosaccharomyces pombe ...
Embodiment 3
[0039] Example 3: Determination of ethanol and fusel oil produced by bacterial strain fermentation
[0040] Taking 2 strains of wine-producing yeast used in a winery as a comparison, the yeasts with different high-yield ethanol and low fusel oil yields provided by this patent were used with a pH of 5 and a sugar content of 10 o Bx Sorghum Juice Medium, 1 x 10 6 See Table 2 for the inoculum size per mL, 30°C, 150 rpm, shake flask culture for 72-96 hours, ethanol and fusel oil concentrations.
[0041] Table 2 Results of ethanol production by fermentation of yeast sorghum juice medium
[0042]
[0043] Note: The theoretical conversion rate of ethanol to sugar is 51.1%.
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