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Method for screening yeast CGMCC 4746 capable of producing ethanol in high yield and producing fusel oil in low yield in production of maotai-flavor Chinese distilled spirits and application of method

A fusel oil and ethanol technology, applied in the field of bioengineering, can solve problems such as numerous influencing factors, production risks, complex processes, etc., and achieve the effects of reducing production costs, improving wine quality, and optimizing production processes

Inactive Publication Date: 2012-11-07
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to its complex process and various influencing factors, it has not been well resolved in production.
The most fundamental measure is to use yeast strains with low fusel oil production for fermentation. However, fusel oil is an inevitable by-product of yeast in the ethanol fermentation process. Strains with high ethanol production capacity generally have a stronger ability to produce fusel oil. Therefore, It is often difficult to have both high-yield ethanol and low-yield fusel oil
At present, there are also reports of alcoholic yeast strains with low fusel oil production through mutagenesis, but there are problems such as back mutations in the brewing industry, and there are risks in production

Method used

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  • Method for screening yeast CGMCC 4746 capable of producing ethanol in high yield and producing fusel oil in low yield in production of maotai-flavor Chinese distilled spirits and application of method
  • Method for screening yeast CGMCC 4746 capable of producing ethanol in high yield and producing fusel oil in low yield in production of maotai-flavor Chinese distilled spirits and application of method
  • Method for screening yeast CGMCC 4746 capable of producing ethanol in high yield and producing fusel oil in low yield in production of maotai-flavor Chinese distilled spirits and application of method

Examples

Experimental program
Comparison scheme
Effect test

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] Numbering Chinese name English name Deposit number 1 Pichia anomalies Pichia anomala CGMCC No.4740 2 Schizosaccharomyces pombe Schizosaccharomyces pombe CGMC...

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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Abstract

The invention relates to a method for screening yeast CGMCC 4746 capable of producing ethanol in high yield and producing fusel oil in low yield in the production of maotai-flavor Chinese distilled spirits and application of the method, and belongs to the technical field of biological engineering. According to the yeast, a strain is trichosporon asahii CGMCC 4746 and is separated from various massive raw starters for alcoholic liquor, fermented grains, brewing raw materials and brewing environmental matters of multiple famous distilleries in China. The screening method comprises the following steps of: taking starter samples, the fermented grains, the brewing raw materials or the brewing environmental matters, diluting, and coating to a triphenyl tetrazole hydrochloride (TTC) preliminary screening flat plate; selecting bacterial colonies with an obvious red color, transferring the red bacterial colonies to a shake flask for culturing, and screening repeatedly; and measuring the content of the ethanol and the fusel oil in fermentation liquor respectively by a distillation specific gravity method and using a headspace-solid-phase micro-extraction-gas chromatography-mass spectrometer (HS-SPME-GC-MS), and screening to obtain the strain with high ethanol concentration and low fusel oil concentration. The strain can be applied to industries of foods and brewed liquors.

Description

technical field [0001] The invention relates to a method for screening yeasts with high ethanol and low fusel oil yields, and the application of the method to screen yeasts with high ethanol and low fusel oil yields in the production of Chinese Maotai-flavor liquor and its application in food technology and brewing wine industry. It belongs to the technical field of bioengineering. Background technique [0002] Ethanol is one of the most important and main components in brewing wine. The ethanol content in Chinese liquor is as high as 40%-55% (v / v); in rice wine, it is 14%-20%; in beer, it is 2%-5%. Yeast is the main microorganism that produces ethanol, and it is also an indispensable microorganism in the brewing industry. Therefore, finding high-yielding yeast can optimize the brewing process and reduce production costs, which is of great significance in the brewing industry. [0003] Fusel oil is a secondary metabolite produced by yeast during brewing fermentation, inclu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16C12G3/02C12G3/12C12G3/04C12R1/645C12H6/02
Inventor 徐岩吴群陈良强
Owner JIANGNAN UNIV
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