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A high-yield ethyl acetate yeast strain under low temperature conditions and its application

A low-temperature condition and ethyl acetate technology, applied in the field of bioengineering, can solve the problems of many types of metabolites, high fermentation temperature, and high production cost, so as to reduce fermentation temperature, increase ethyl acetate content, and reduce production energy consumption and cost Effect

Active Publication Date: 2018-07-13
SICHUAN UNIVERSITY OF SCIENCE AND ENGINEERING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the problems of high process difficulty, increased energy consumption, high production cost, and many types of metabolites caused by high fermentation temperature in the production process of fermented food, the present invention provides an acid-resistant, salt-resistant, ethanol-resistant, low-temperature conditioner That is, the saccharomycete that can produce ethyl acetate at a high rate, the object of the present invention is achieved by the following technical solutions:

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The screening of embodiment 1 ester-producing yeast

[0020] Grind the koji medicine and fermented unstrained spirits with a mortar, weigh 10g to 100ml of sterile normal saline, and vibrate at 150r / min for 30min to obtain the bacterial suspension, perform ten-fold serial dilution, and spread the dilution on the primary screening culture Base, select the ratio of the diameter of the transparent circle and the diameter of the colony to be a large colony, that is, a strain with significant ester production. Then transfer the strain, carry out shake flask fermentation and re-screening, use HS-SPME-GC-MS method to determine the type of ester in the fermentation liquid, use the method for determining the total ester of liquor stipulated in GB / T 10345-2007 to determine the content of ethyl acetate in the fermentation liquid , and screened to obtain strains with strong ability to produce ethyl acetate.

[0021] The initial screening culture conditions for ester production are:...

Embodiment 2

[0027] Example 2 Ethyl acetate-producing yeast and its molecular biology identification

[0028] The obtained ethyl acetate-producing yeast was identified by molecular biology, 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 strain was deposited in the General Microorganism Center of China Committee for Culture Collection of Microorganisms, and the yeast species and preservation number were: Wickerhamomycesanomalus (Wickerhamomycesanomalus) CGMCC NO: 10370.

[0029] The process of confirming the species of yeast is as follows:

[0030] Identification of 26SrDNA sequence homology of isolated strains: Extraction of yeast DNA: inoculate the yeast in liquid YPD medium after activation, and culture it on a shaker at 28°C for 12 hours, then take a small amount of bacteria into a ste...

Embodiment 3

[0035] The acid resistance test of embodiment 3 yeast strains

[0036] Adjust the pH of the YPD liquid medium to 1.0-7.0, increase the gradient with pH 0.5, do three parallels for each gradient, inoculate 10% (V / V) yeast suspension, at 30°C, 150r / min, after culturing for 24h, Use a microplate reader to measure the turbidity of the culture medium at a wavelength of 600nm to determine the growth of the bacteria. The acid resistance results of the strains are shown in Table 1 below.

[0037] Table 1 strain acid resistance (OD 600 value)

[0038] pH

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

OD 600

0.002

1.085

1.291

1.341

1.335

1.403

1.403

1.425

1.421

1.417

1.433

[0039] Table 1 shows the OD of the strains at pH 2.5 600 reached 1.085, indicating that the strain can grow normally under the environment of pH2.5,

[0040] And with the increase of pH, the strain can still grow well in acidi...

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Abstract

The invention discloses a strain of yeast with high ethyl acetate production under low temperature conditions, which belongs to the technical field of bioengineering. The strain is Wickerhamomycesano malus, which was deposited in the General Microorganism Center of China Committee for the Collection of Microorganisms on January 19, 2015, with the preservation number CGMCC NO.10370. The optimum temperature of bacterial strain producing ethyl acetate of the present invention is 22 ℃, and acid resistance is pH2.5~7.0; Salt tolerance is 0~15% (NaCl%); Ethanol resistance is 0~10% (v% ), the metabolites produced by fermentation are only ethyl acetate and a small amount of ethanol, and do not contain fusel oil. The bacterial strain of the present invention can be used as an important functional bacterium in traditional fermentation industries and food industries such as vinegar, soy sauce, tempeh, noodle sauce, brewed wine, distilled wine, and prepared wine, so as to reduce the fermentation temperature of traditional production, optimize the production process, and reduce production. Energy consumption and cost are reduced, while increasing the content of ethyl acetate in the product, improving the flavor of the product, improving the quality of the product, and having considerable economic benefits.

Description

technical field [0001] This patent belongs to the technical field of bioengineering, and specifically relates to a high-yield ethyl acetate-producing yeast under low temperature conditions and its application. Background technique [0002] Ester-producing yeast, also known as aromatic yeast, is not a term in yeast taxonomy, but a general term for yeast that can produce a large amount of ester substances. Saccharomyces and Brettanomyces etc. Esters belong to an organic compound, and lower esters are a class of volatile liquids with aroma. Most of the ester-producing yeasts are some wild yeasts, they are widely distributed in nature, and they are reflected in different degrees in the raw materials, manufacturing processes and finished products of the brewing industry. They occupy a pivotal position in my country's brewing industry and are widely used It is used in traditional fermented foods such as white wine, rice wine, wine, soy sauce, watercress, sweet noodle sauce, and v...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16C12P7/62C12R1/645
CPCC12P7/62C12N1/145C12R2001/645
Inventor 黄丹毛祥罗惠波叶光斌刘有晴于华方春玉
Owner SICHUAN UNIVERSITY OF SCIENCE AND ENGINEERING
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