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Method for high throughput screening of saccharomyces cerevisiae

A beer yeast and purpose technology, applied in the field of high-throughput screening of beer yeast, can solve the problems of slow screening work, heavy workload, and limited number of screenings, and achieve the effects of reducing screening work intensity, improving efficiency, and increasing the number of colonies

Inactive Publication Date: 2016-04-06
BEIJING YANJING BREWERY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the simple operation of shaking flask primary screening, but the workload is large, and human error is also large, and the number of each screening is limited, and the screening speed is slow. Therefore, it is necessary to establish a set of high-throughput screening models belonging to brewer's yeast strains.

Method used

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  • Method for high throughput screening of saccharomyces cerevisiae
  • Method for high throughput screening of saccharomyces cerevisiae
  • Method for high throughput screening of saccharomyces cerevisiae

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0033] The preparation method of the wort medium in the following examples: the wort (Yanjing Brewery Co., Ltd.) was sterilized and obtained.

[0034] The preparation method of the wort agar medium in the following examples: add 2% agar powder to the liquid wort medium.

[0035] The deposit number of brewer's yeast strain 8902 in the following examples is CGMCC No.1784.

[0036] The preservation number of brewer's yeast strain 0002 in the following examples is CGMCC No.10572.

[0037] The brewer's yeast strain 68obg in the following examples is a product of the Berlin Institute of Brewing, Germany, product number: 016719.

[0038] The brewer's yeast strain O.K.3 in the following examples is a product of the Berlin Institute of Brewing, Germany, product number: 016720.

Embodiment 1

[0039] Embodiment 1, the optimization of brewer's yeast culture parameter

[0040] 1. Parameter optimization

[0041] 1. Treatment of strains

[0042] The brewer's yeast strain (8902CGMCCNo.1784) to be screened is activated and expanded to obtain a cultured bacterial suspension after activation. The specific steps are as follows:

[0043] (1) Use an inoculation loop to pick up a loop of the strains stored on the slant and add it to a test tube containing 10mL of 12°P wort medium, and incubate at 25±1°C for 24 hours.

[0044] (2) Gently oscillate the culture solution in the test tube after the cultivation, and add 1 mL of it to a test tube containing 10 mL of 12°P wort medium with a sterile pipette, and incubate at 25±1°C for 24 hours after oscillating evenly.

[0045] (3) Gently oscillate the test tube culture solution after the end of the culture, and then put them all into a small Erlenmeyer flask filled with 100mL of 12°P wort medium, and incubate at 25±1°C for 24 hours a...

Embodiment 3

[0064] Embodiment 3, the application of the method for high-throughput screening brewer's yeast in screening brewer's yeast

[0065] 1, Erlenmeyer flask fermentation method (contrast)

[0066] In this example, the traditional shaking flask screening (triangular flask fermentation method) was used as a comparison, and the brewer's yeast strains (8902, 68obg, 0002 and O.K.3) were fermented and cultured, and the brewer's yeast was screened according to the diacetyl content or OD value in the fermentation broth strain. Specific steps are as follows:

[0067] (1) The brewer's yeast strains (8902, 68obg, 0002 and O.K.3) were activated and expanded (see step 1 in Example 1 for the expansion method), and the activated bacterial suspensions were respectively obtained.

[0068] (2) Fermentation culture

[0069] Inoculate the bacterial suspension obtained in the above step (1) into the Erlenmeyer flasks equipped with 350mL wort medium (12°P) for fermentation and culture respectively, ...

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Abstract

The invention discloses a method for high throughput screening of saccharomyces cerevisiae. The screening method is improved on the basis of a conventional saccharomyces cerevisiae screening method; micro-porous cell culture plates are used for replacing test tubes or shake flasks in screening; 24-384 samples can be cultured by one micro-porous cell culture plate, and simultaneously shaker fermentation is conducted; a plurality of the micro-porous cell culture plates can be placed on a shaker; and preliminarily screened products, separated from natural world, or products of artificial mutagenesis such as ultraviolet mutagenesis can be used as to-be-screened raw materials of the method. By virtue of the method disclosed by the invention, the colony number in each screening is effectively increased, the working strength of screening is reduced and the efficiency of the entire screening work is improved; and the method has a positive effect on the breeding of saccharomyces cerevisiae strains.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for high-throughput screening of brewer's yeast. Background technique [0002] High-throughput screening (HTS) technology is based on experimental methods at the molecular and cellular levels, using microplates as the carrier of experimental tools, using automated operating systems to execute the experimental process, and collecting experimental results with sensitive and rapid detection instruments. Data, the experimental data is analyzed and processed by computer, tens of millions of samples are tested at the same time, and the corresponding database is used to support the operation of the technical system. At present, this technology is relatively mature in foreign countries. With the development of science and technology, a variety of fully automatic high-throughput screening systems have appeared one after another. The automatic high-throughput screening sy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12N1/18C12R1/865
Inventor 郭立芸朱佳静穆英健梁云林智平贾凤超
Owner BEIJING YANJING BREWERY
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