A low-temperature resistant yeast jm33 and its application in Munage wine
A technology of Munag wine and Munag grapes, which is applied to Saccharomyces cerevisiae and applied in wine brewing, food microorganism application field, and achieves the effects of pleasant aroma, high ethanol yield and unique flavor.
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Embodiment 1
[0034] Example 1: Isolation, screening and identification of Saccharomyces cerevisiae JM33 CGMCC NO.10912
[0035] 1. Isolation and screening of strains
[0036] The Saccharomyces cerevisiae used in the present invention is sampled and separated from mature Munage grapes, and according to the analysis of the action mechanism of the strains, the bacterial strains with a certain fermentative ability to Munage grapes are preliminarily screened out. The microorganisms in Munag grape juice were isolated by traditional plate culture method, the strains were screened by primary screening and secondary screening, and the bacterial strains were purified by plate streaking method, and the sugar resistance, alcohol resistance, acid resistance, sulfur dioxide resistance, and sulfur dioxide resistance of the strains were compared. Potassium chloride resistance and low temperature resistance, a batch of well-growing microbial strains were screened out, and a Saccharomyces cerevisiae strain ...
Embodiment 2
[0055] Example 2: Molecular determination of Saccharomyces cerevisiae JM33 CGMCC NO.10912
[0056] 1. PCR amplification of the DNA sequence of Saccharomyces cerevisiae and its sequencing
[0057] Pick a small amount of single colonies of the JM33 strain, put them into an EP tube filled with 25 μL of sterile water, boil at 100°C for 8-10 minutes, and then quickly put them into the ice-water mixture for 5 minutes. Centrifuge at 10000r / min for 5min, store at 4°C, and take the supernatant when used.
[0058] Determination of 26S rDNA gene sequence and construction of phylogenetic tree: extract the total DNA of yeast strains according to conventional methods, and use deionized water to perform PCR amplification of the 26S rDNA segment with diluted universal primers NL1 and NL4. The primers are designed as follows :
[0059] NL1: 5'-GCATATCAATAAGCGGAGGAAAAG-3'
[0060] NL4: 5'-GGTCCGTGTTTCAAGACGG-3'
[0061] The 50 μl reaction system contains: 5 μl of 10×PCR buffer, 20 pmol of e...
Embodiment 3
[0064] Embodiment three: Utilize Saccharomyces cerevisiae (Saccharomyces cerevisiae) JM33 CGMCC NO.10912 and Mu Nage grape to make wine
[0065] (1) Inoculation: Prepare the YPD solid medium of Saccharomyces cerevisiae JM33 CGMCC NO.10912, perform strict aseptic operation after sterilization, transfer from the slant to the plate, and culture at 28°C for 2 days.
[0066] (2) Expansion culture: Pick a single colony from the YPD solid medium in step (1) and transfer it to an Erlenmeyer flask containing YPD liquid medium, perform aseptic operation, and cultivate at 28°C and 120r / min for 24h- After 48 hours, the seed solution was prepared.
[0067] (3) Collection of seed liquid: the seed liquid activated in step (2) was centrifuged and washed twice and then collected for later use. The centrifugation speed was 5000 rpm.
[0068] (4) Selection of Munage grapes: select ripe Munage grapes without mildew and insect damage, and reserve them for later use.
[0069] (5) Processing of ra...
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