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Pichia kudriavzevii with high tolerance and low isoamyl alcohol yield and application of pichia kudriavzevii

A technology of Pichia pastoris and isoamyl alcohol, applied in the field of brewing microorganisms, can solve problems such as unpleasantness, and achieve the effects of improving wine quality, unique high-yield ethanol and low-yield fusel oil, and excellent physiological tolerance

Pending Publication Date: 2022-08-02
HUBEI DAOHUAXIANG WINE IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the wine body, an appropriate amount of higher alcohol can balance the taste and aroma, but studies have shown that too much higher alcohol in the wine body will not only bring bitterness and unpleasant "fusel alcohol taste", but also after drinking Can cause severe alcohol stress effects such as headache, nausea, and palpitations

Method used

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  • Pichia kudriavzevii with high tolerance and low isoamyl alcohol yield and application of pichia kudriavzevii
  • Pichia kudriavzevii with high tolerance and low isoamyl alcohol yield and application of pichia kudriavzevii
  • Pichia kudriavzevii with high tolerance and low isoamyl alcohol yield and application of pichia kudriavzevii

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Experimental program
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Effect test

Embodiment 1

[0023] Screening of strains

[0024] The Pichia pastoris with high ethanol yield and low fusel oil yield involved in the present invention is isolated and screened from high-quality fermented grains, and the specific screening method is as follows:

[0025] First, more than 60 yeast strains with excellent performance were screened out of high-quality fermented grains, and directional screening was carried out for ethanol-producing and low-yielding fusel oil. , prepare multiple plates in advance, and then inoculate all strains on the plate for two days at 30°C, pour the TTC medium cooled to 45°C into the upper layer of the yeast extract peptone dextrose medium (YEPD) plate and store it in the dark for two days. Observe the color change after an hour, and select the strain with darker red as the alternative strain (specifically numbered as Shao, Fen II, PD1, PT11, PT17, Y-3, JY26, PF4, M3, M4, 4-16, 2300, 1312 , 10#). Then the alternative strain was placed in yeast extract pep...

Embodiment 2

[0030] Example 2: Physiological characteristics of Pichia pastoris with high ethanol production and low fusel oil production:

[0031] Scrape FenⅡ, PT11, JY26, 2300, 1312, M3 from a ring of slants to liquid yeast extract peptone glucose medium (YEPD) at 30°C, 180r / min for 16h, and then inoculate YEPD liquid according to 2% of the inoculum The medium was cultured at 30°C at 180r / min for 24h, and the OD600 was used as an indicator to detect the ethanol concentration of the strain at 1%, 3%, 5%, 7%, 9%, 11%, 13%; pH value is 2, 3, 4, 5, 6, 7, 8; Brix value is 5Brix, 10Brix, 15Brix, 20Brix, 25Brix, 30Brix, 35Brix; culture temperature is 18°C, 22°C, 26°C, 30°C, 34°C ℃, 38℃, 42℃; lactic acid concentration: 0%, 1%, 2%, 3%, 4%, 5%, caproic acid concentration: 0‰, 0.2‰, 0.4‰, 0.6‰, 0.8‰, 1.0‰; the acetic acid concentration is: tolerance under the conditions of 0‰, 1‰, 2‰, 3‰, 4‰, 5‰, and the results are shown in the appendix figure 1 . It can be seen that the M3 strain has the best ...

Embodiment 3

[0036] Example 3: Molecular biological identification of M3 strain

[0037]The strains obtained by screening were sent to Huada Gene Technology Co., Ltd. for 18S rRNA gene sequencing. The sequencing results were placed in the NCBI (National Center for Biotechnology Information) database for comparison, and the strain sequences with higher similarity were selected and used in MEGA 11.0 software. Maximum likelihood method (Maximum Likelihood Estimation, MLE) to construct phylogenetic tree as attached Figure 4 shown.

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Abstract

The invention provides a pichia kudriavzevii strain with high tolerance and low yield of isoamyl alcohol and application of the pichia kudriavzevii strain, the pichia kudriavzevii strain is pichia kudriavzevii strain M3, and the preservation number of the pichia kudriavzevii strain is CCTCC (China Center for Type Culture Collection) NO: M 2022254. The yeast has the capacity of high tolerance, high ethanol yield and low fusel oil yield, the bacterial colony of the yeast is round, the edge is zigzag, and the shape is large. The bacterial strain is slightly bulged, is milk white in appearance and is dry, the cell morphological characteristic of the bacterial strain observed under a 400-fold microscope is elliptical or prolate, and the propagation mode is budding. The invention also relates to application of the pichia pastoris in the field of wine brewing and / or yeast preparations. The invention further provides liquor for blending in fen-flavor liquor, the liquor is blended with base liquor, the purposes of improving the purity and the superior product rate of the liquor can be achieved, and the high-quality liquor which is pure, mellow, sweet and free of drunkenness is obtained.

Description

technical field [0001] The invention relates to the field of brewing microorganisms, in particular to a Pichia strain with good tolerance, high ethanol yield and low fusel oil yield and application thereof. Background technique [0002] Higher alcohols refer to the general term for monohydric alcohols containing three or more carbon atoms, including n-propanol, isobutanol, isoamyl alcohol, etc.; The main microorganisms that produce higher alcohols in the early stage of fermentation have sufficient substrates, yeast proliferate in large quantities, and higher alcohols are primary metabolites, so they are mainly generated at this stage; and in the later stage of fermentation, some of these higher alcohols will be mixed with acetic acid. The acid substances are catalyzed by enzymes to generate corresponding ester compounds, and some of them continue to exist in the wine body. In the wine body, a moderate amount of higher alcohol has the effect of balancing the taste and aroma,...

Claims

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Application Information

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IPC IPC(8): C12N1/16C12G3/021C12G3/04C12R1/84
CPCC12N1/165C12N1/16C12G3/021C12G3/04C12R2001/84Y02E50/10
Inventor 蔡开云陈萍陈小林冯向东万小丰明聃杨博郭婷婷郑裴
Owner HUBEI DAOHUAXIANG WINE IND
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