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A technology of wine yeast and higher alcohol, applied in the field of wine yeast, can solve the problem of high content of higher alcohol, achieve the effect of reducing the content and improving the health value
Inactive Publication Date: 2014-03-12
SHANDONG AGRICULTURAL UNIVERSITY
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[0005] The purpose of the present invention is to provide a strain of wine yeast with low yield of higher alcohol, and provide a fast, accurate and convenient method for breeding yeast with low yield of higher alcohol. Utilizing the yeast to produce wine can solve the problem of high alcohol content in wine And make consumers get headaches and headaches after drinking
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Embodiment 1
[0032] Embodiment 1: Breeding of low-yielding high-alcohol wine yeast
[0033] 1) Mutagenesis of the starting strain:
[0034] a. Pick a small amount of yeast cells from the slant of wine yeast EC1118 with an inoculation needle as the starting strain, inoculate it into 20 mL of YPD liquid medium, and culture it with shaking at 30°C for 16-18 hours to obtain the starting strain liquid.
[0035] b. Then take 1mL of the starting strain bacterial solution in a centrifuge tube at 12000r / min, centrifuge for 15s to collect the cells, and wash once with sterile water.
[0036] c. Suspend the cells with 1 mL of sterile sodium phosphate buffer (a mixture of sodium dihydrogen phosphate, disodium hydrogen phosphate and sterile water; the concentration of the buffer is 0.1mol / L) with a pH value of 7.0, and add 30 μL of methanesulfonic acid Ethyl ester (EMS) stock solution, shake to disperse the cells, place in a shaker at 30°C for 1 hour and then centrifuge.
[0037] d. Then add a 5% ste...
Embodiment 2
[0044] Example 2: Analysis of higher alcohol production of the starting strain and the target strain
[0045]Extraction of higher alcohols: Accurately measure 8mL of wine fermented by strain CP1118, put it into a 15mL headspace bottle, add 1g NaCl to promote the volatilization of higher alcohols, and then add 30μL of internal standard (4-methyl-2-pentanol) , and immediately seal and compress with a PTFE / silicone rubber septum. When the wine sample temperature reached 45°C, it was extracted with DVB / CAR / PDMS extraction head for 50 minutes, and the sample was injected for gas chromatography-mass spectrometry analysis.
[0046] GC / MS conditions: chromatographic column, Stabilwax-DA capillary column ((30m×0.32mm×0.25μm)); heating program: keep at 30°C for 1min, rise to 100°C at 6°C / min, and raise temperature at 3°C / min To 200°C, raise the temperature to 210°C at 10°C / min, and keep for 3min; the injector temperature is 250°C; the detector temperature is 250°C, splitless injection....
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Abstract
The invention relates to wine yeast with low yield of higher alcohol. The wine yeast is characterized in that a certain amount of isoamyl chloroacetate is added in a flat plate for cultivating yeast, the isoamyl chloroacetate generates chloroacetic acid under the catalysis of saccharomycetes isoamyl acetate hydrolase; the chloroacetic acid has a strong restraining effect on the growth of saccharomycetes, if the activity of the ester decomposition enzyme of a mutant strain is high (the higher the activity of the ester decomposition enzyme is, the more higher alcohol produced by the strain is), more chloroacetic acid is produced by decomposition, so that a bacterial colony grows slowly under the inhibition effect of the chloroacetic acid. The wine yeast can be utilized to accurately, conveniently and quickly screen out saccharomycetes with low yield of the higher alcohol. When the saccharomycetes with low yield of the higher alcohol, which is bred through the wine yeast, is used for producing grape wine, the content of the higher alcohol in the grape wine can be lowered by about 10% to 15%; when the saccharomycetes with low yield of the higher alcohol, which is bred through the wine yeast, is used for alcohol fermentation, the content of the higher alcohol in grape wine can be lowered effectively, and the health-care values of the grape wine can be improved.
Description
(1) Technical field [0001] The invention relates to a strain of wine yeast with low yield of higher alcohol, which belongs to the field of wine brewing. (2) Background technology [0002] Higher alcohols are the most common yeast secondary metabolites in wine. If the content of wine is too high, it will cause consumers to feel dizzy, headache and drunk after drinking, which will seriously affect the health effect of wine. [0003] The higher alcohols produced by wine yeast metabolism mainly include isoamyl alcohol, isobutanol, n-propanol and isohexyl alcohol, among which isoamyl alcohol accounts for about 40% to 50% of the composition of wine higher alcohols. Existing studies have mainly used the diazo staining plate method to screen low-yielding yeast strains to reduce the content of higher alcohol in wine. Diazo staining plate color development principle: yeast isoamyl acetate hydrolase catalyzes 1-naphthyl acetate on the plate to generate naphthol, which reacts with azo ...
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