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Yeast strain for highly yielding ethyl acetate, and culture method and application thereof

A yeast strain and ethyl acetate technology, applied in the field of microorganisms, can solve the problems of difficult application to actual production, poor tolerance, low ester production level, etc., and achieve optimized components and culture conditions, high ethanol tolerance, and improved yield. Effect

Active Publication Date: 2018-05-11
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are a variety of ester-producing yeast strains, there are still problems such as low ester production level and poor tolerance, which are difficult to apply to actual production

Method used

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  • Yeast strain for highly yielding ethyl acetate, and culture method and application thereof
  • Yeast strain for highly yielding ethyl acetate, and culture method and application thereof
  • Yeast strain for highly yielding ethyl acetate, and culture method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Abnormal Wickham yeast ( Wickerhamomyces anomalus ) Isolation of Y3604

[0045] After pulverizing Gujing Gong Daqu and mixing it evenly, weigh 1 g, add 9 mL of sterile water, oscillate fully and soak for 15 min to make a suspension. Under aseptic conditions, take 0.1 mL of the suspension and dilute it step by step with sterile water to 10 -3 、10 -4 、10 -5 、10 -6 . Take 0.1 mL of each gradient suspension on a YPD plate, make three parallels for each dilution gradient, and spread sequentially from low concentration to high concentration. Place the medium plate at 30 o C cultured in a constant temperature incubator for 2 days to observe the growth of colonies. Pick up a single colony that is spherical, protruding on the surface, milky white, and opaque on the plate, and inoculate it on the YPD plate for multiple times until the bacteria are in the same shape under the microscope. A single colony was inoculated in 50 mL of sorghum extract medium, 30 o C....

Embodiment 2

[0046] Example 2 Abnormal Wickham yeast ( Wickerhamomyces anomalus ) Y3604 save

[0047] The above-mentioned abnormal Wickerhammyces anomalus (Wickerhamomyces anomalus) Y3604 was preserved by the following method:

[0048] (1) Preservation on slant: Inoculate the purified strains on the YPD slant, place it in the incubator for 48-72 h, and then store it at 4 o C stored in the refrigerator.

[0049] (2) Storage in glycerol tubes: Take 6 mL of 20% glycerol on the purified strain plate, scrape off the colony and mix it with 20% glycerol, divide the mixture into 1.5 mL sterile PE tubes, store at -80 o Save under C.

Embodiment 3

[0050] Example 3 Abnormal Wickham yeast ( Wickerhamomyces anomalus ) Identification of Y3604

[0051] The abnormal Wickham yeast involved above ( Wickerhamomyces anomalus ) The identification of Y3604 includes the following steps:

[0052] Step 1: Morphological Characterization

[0053] In order to identify the yeast strains involved, the following morphological characteristics were observed:

[0054] (1) Colony morphology and cell observation in YPD medium: the pure culture of the strain obtained in Example 1 was inoculated on YPD medium, and its morphological characteristics were observed after 48 h ( figure 1 ), the results are shown in Table 1, and observed under an optical microscope after being magnified 400 times, the results are as follows figure 2 , oval in shape, budding.

[0055] (2) Observation of colony morphology in WL medium: inoculate the screened strains on YPD for activation, culture for 24-48 h and then inoculate into WL medium, 30 o C cultured and...

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Abstract

The invention relates to a Wickerhamomyces anomalus strain for highly yielding ethyl acetate, and a culture method and an application thereof. The strain for highly yielding ethyl acetate is named asa Wickerhamomyces anomalus Y3604 strain, and is preserved in China General Microbiological Culture Collection Center on October 12, 2016 with the preservation number of CGMCC No.13103; and the 26S rDNA D1 / D2 sequence of the strain is similar to the 26S rDNA D1 / D2 sequences of multiple other Wickerhamomyces anomalus strains. The strain has good tolerance to glucose, ethanol and ethyl acetate, and has a wide range in the growth pH value and the temperature; and the Wickerhamomyces anomalus Y3604 strain has a high ethyl acetate yield, and a result of detection shows that the ethyl acetate yield of the strain can reach 19.17 g / L. The strain can be applied to the brewing industry of Chinese liquor, yellow rice wine and soy sauce, demanding for the ethyl acetate.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a newly isolated high-yield ethyl acetate yeast strain and its cultivation method and application. Background technique [0002] Ethyl acetate is a liquid with fruity aromas such as apple and pineapple, and its production mainly depends on the balance between the synthesis of esters and the hydrolysis of esters. Ethyl acetate is widely used in the food industry, and the food industry has a high demand for ethyl acetate, so it is of far-reaching significance to seek a cheap and safe way to generate ethyl acetate. At present, there are three main ways to produce ethyl acetate: one is to synthesize ethyl acetate through chemical methods, and then purify it, which has certain risks; the other is the fermentation of microorganisms, mainly ester-producing yeast such as Pichia anomaly The ethyl acetate can be regarded as natural ethyl acetate; the third is to extract from ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16C12P7/62C12G3/02C12R1/645
CPCC12G3/02C12P7/62C12N1/145C12R2001/645
Inventor 李秀婷范光森孙宝国滕超许岱富志磊
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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