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Saccharomycete for high-yield production of ethyl acetate under low temperature and application thereof

A low-temperature condition, ethyl acetate technology, applied in the field of bioengineering, can solve the problems of many types of metabolites, high fermentation temperature, difficult process, etc., and achieve the effects of reducing fermentation temperature, increasing ethyl acetate content, and considerable economic benefits.

Active Publication Date: 2016-02-24
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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  • Saccharomycete for high-yield production of ethyl acetate under low temperature and application thereof
  • Saccharomycete for high-yield production of ethyl acetate under low temperature and application thereof
  • Saccharomycete for high-yield production of ethyl acetate under low temperature and application thereof

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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 the bacterial strain was transferred, and the shake flask fermentation was re-screened. The ester type of the fermentation liquid was determined by the HS-SPME-GC-MS method, and the ethyl acetate content of the fermentation liquid was determined by the method for determining the total esters of liquor stipulated in GB / T10345-2007. The strains with strong ability of producing ethyl acetate were screened.

[0021] The initial screening culture conditions for ester p...

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) CGMCCNO: 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 after cultivating on a shaker at 28°C for 12 hours, take a small amount of bacteria into a st...

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 with a gradient of pH0.5, 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 under the condition of 600nm wavelength 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 pH2.5 600 reached 1.085, indicating that the strain can grow normally under the environment of pH2.5,

[0040] And with the increase of pH, ...

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Abstract

The invention discloses saccharomycete for high-yield production of ethyl acetate under low temperature and belongs to the field of the bioengineering technology. The bacterial strain is Wickerhamomycesanomalus which is preserved in the China General Microbiological Culture Collection Center on Jan 19, 2015 with the preservation number being CGMCC No.10370. The optimum temperature for producing ethyl acetate by means of the strain is 22 DEG C, acid resistance is pH 2.5-7.0, salt tolerance is 0-15% (NaCl%), and ethanol tolerance is 0-10%(v%). Metabolites generated from fermentation are ethyl acetate and a small amount of ethyl alcohol only and do not include fusel oil. The strain can be applied to the traditional fermentation industry for producing table vinegar, soy sauce, fermented soya beans, fermented flour sauce, fermented alcoholic drink, distilled liquor, compound wine and the like, as well as the food industry as an important functional bacterium. Fermentation temperature adopted in traditional production is reduced, the production technology is optimized, production energy consumption and cost are reduced, the content of ethyl acetate is increased, the taste of a product is improved, the quality of the product is improved, and economic benefits are great.

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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IPC IPC(8): C12N1/16C12P7/62C12R1/645
CPCC12P7/62C12N1/145C12R2001/645
Inventor 黄丹毛祥罗慧波叶光斌刘有晴于华方春玉
Owner SICHUAN UNIVERSITY OF SCIENCE AND ENGINEERING
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