Screening method of grape wine yeast with low-yield ethyl carbamate and application thereof

A technology of urethane and wine yeast, which is applied in the preparation of wine, the preparation of alcoholic beverages, methods based on microorganisms, etc., can solve the problems of no successful case reports of breeding

Inactive Publication Date: 2011-10-19
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

France and Argentina have completed O.oeni and L. hilgardii The full sequence of the arc gene cluster will provide

Method used

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  • Screening method of grape wine yeast with low-yield ethyl carbamate and application thereof
  • Screening method of grape wine yeast with low-yield ethyl carbamate and application thereof
  • Screening method of grape wine yeast with low-yield ethyl carbamate and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] 1. Activation of Strains

[0087] Take 0.2 g of commercially available F15 active dry yeast, add it to a Erlenmeyer flask filled with 10 mL of 5% glucose solution, and activate it in a water bath at 38-40 °C for 20-30 min until a large number of bubbles are generated.

[0088] 2. Spread 0.1 mL of the activated dry yeast solution on the mixed medium and culture it at 30°C for three days. Figure 6 Shown:

[0089] 3. Pick out the single colonies A1 and A2 that grew on the mixed medium and put them into a test tube filled with sterile water, and then spread 0.1 mL of the bacterial solution on the mixed medium to isolate the single colonies by dilution and separation. separate.

[0090] 4. In order to prevent the strains screened out from the CAN1 mutation, it needs to be verified. Inoculate the single colony grown on the mixed medium on the ornithine medium and the arginine medium respectively, if it does not grow or grows weakly on the arginine medium but grows on the ...

Embodiment 2

[0093] 1. Isolation of Spontaneously Fermenting Yeast

[0094] Collect grape berries at multiple points in the vineyard, remove stems and crush them, put them into sterilized or sulfur-fumigated containers for natural fermentation, inoculate once every three days from vigorous fermentation to the end of fermentation, absorb 0.1mL and spread on YEPD (yeast Complete culture medium.) culture medium, cultured in a 30°C incubator for 2-3 days, and picked a single colony for preservation.

[0095] 2. Screening of WL nutrient medium

[0096] WL Nutrient Medium is designed to monitor microbial populations during beverage fermentation. Studies have shown that most of the typical yeast species that appear during natural wine fermentation can be distinguished by WL nutrient medium, mainly based on colony color and colony shape. The colony color and morphology of Saccharomyces cerevisiae growing on WL nutrient medium are as follows: cream color with green, spherical protrusions, smooth ...

Embodiment 3

[0102] 1. Activation of Strains

[0103] Take the bacterial lawn on the slope of F-15-A1 and put it into a Erlenmeyer flask filled with 10mL of 5% glucose solution, and activate it in a water bath at 38-40°C for 20-30min until a large number of bubbles are generated.

[0104]2. Ultraviolet-acridine orange mutagenesis: Take the well-grown F-15-A1 seed solution, irradiate with ultraviolet light for 90s, and then transfer 600μL of the bacterial suspension undergoing ultraviolet mutagenesis into a test tube containing 5mL of mutagenesis medium Incubate in a dark water bath for 12 hours at 30°C with shaking. (The mutagenesis medium contains 1mg / mL acridine orange, the doses are 80μL, 100μL, 150μL, 200μL, 250μL);

[0105] 3. Spread 0.1 mL of the yeast liquid after mutagenesis on the mixed medium, and culture it at 30°C for 3 days. Figure 13 Shown: Pick out the single colony B1 (F-15-A1-B1) grown on the mixed medium and put it into a test tube filled with sterile water, and spread...

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Abstract

The invention discloses a screening method of grape wine yeast with low-yield ethyl carbamate and application of grape wine yeast in the production of grape wine. The screening method comprises the following steps of: screening out a bacterial strain with the low-yield ethyl carbamate from active dried yeast and mutant strains by adopting a structural analogue of arginine, i.e., canavanine resistance flat plate, wherein the activity dried yeast can be separated from nature or can be purchased in market, and the mutant strains are obtained through the mutagenesis of chemical mutagen. The method is designed by a yeast amino acid metabolism theory and is simple and convenient; and a target bacterial strain can be obtained through rapid separation. According to the screening method, the obtained resistance bacterial strain B1 can be numbered as F15-1-15 bacterial strain (CGMCC (China General Microbiological Culture Collection Center) NO. 4727) and is applied to the brewing of the grape wine in a small-sized fermented container; and on the basis of excellent brewing performance of the grape wine, the bacterial strain has the advantages of being capable of effectively and obviously reducing the content of the ethyl carbamate in the production of the grape wine and having the potential of industrial production and application.

Description

technical field [0001] The invention belongs to the field of biotechnology food and brewing engineering technology, is an organic combination of modern biological and chemical engineering technology and traditional brewing engineering technology, and creates a wine yeast suitable for wine production by improving the performance of wine yeast used in wine production. A screening method and application of wine yeast that can significantly reduce the content of ethyl carbamate (EC) in wine. Background technique [0002] With the development of my country's economy and the improvement of people's living standards, people pay more and more attention to food health care and quality safety. Wine is a green drink rich in nutrition and health care. Since the 1990s, my country's wine production and consumption have grown rapidly. Wine production increased from 201,900 tons in 2000 to more than 1.2 million tons in 2008, ranking sixth in the world. With the rapid development of the w...

Claims

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Application Information

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IPC IPC(8): C12N1/18C12N13/00C12N15/01C12G1/022C12R1/865
Inventor 王德培高年发李磊张健高强
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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