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A strain of Pichia anomaly with high production of 4-vinylguaiacol and ethyl acetate and its application

A technology of guaiacol and ethyl acetate, applied in the biological field

Active Publication Date: 2019-08-09
ANHUI RUISIWEIER TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biological preparation method of 4-vinyl guaiacol in Chinese patents is based on Bacillus as the experimental strain, and yeast, especially Pichia pastoris, is used as the strain to produce 4-vinyl guaiacol by fermentation. see the report

Method used

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  • A strain of Pichia anomaly with high production of 4-vinylguaiacol and ethyl acetate and its application
  • A strain of Pichia anomaly with high production of 4-vinylguaiacol and ethyl acetate and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The screening of embodiment 1 Pichia anomala GJYL08 bacterial strain

[0025] 1) Take a certain amount of medium-high temperature Daqu from Anhui Gujing Gongjiu Co., Ltd., crush it and pass it through a 40-mesh sieve, add it to an appropriate amount of sterile saline filled with glass beads, and shake it on a shaker at 30°C and 120rpm for 30 minutes to prepare Bacterial suspension; the bacterial suspension was serially diluted and spread on Bengal red medium, and cultured at 28°C for 48h.

[0026] 2) Pick the typical yeast colonies grown on the plate for descriptive records, and make slides for microscopic examination at the same time, select different types of yeast colonies and inoculate them in the sterilized YPD medium (glucose 2g, tryptone 2g) , yeast extract 1g, distilled water 100mL, pH 5.0-5.5, sterilized at 115°C for 20min), and cultivated at 28°C for 48h. Purify until a single colony of the strain is obtained as a primary screening strain.

[0027] 3) Differ...

Embodiment 2

[0031] The liquid state fermentation of embodiment 2 Pichia anomala GJYL08 bacterial strain

[0032] 1) Strain activation: Pick a single colony of Pichia anomala GJYL08 stored in a YPD inclined test tube, inoculate it in a petri dish containing YPD medium, and culture it at 26°C for 32 hours to obtain an activated single colony;

[0033] 2) Bacterial suspension preparation: Pick 2 rings of the above-mentioned activated single colony and inoculate it in the sorghum saccharification liquid medium with a sugar content of 10°BX, culture at 26°C and 120rpm for 24 hours to obtain the bacterial suspension;

[0034] 3) Fermentation broth preparation: the above-mentioned bacterial suspension was used as a strain, inoculated into sorghum saccharification broth medium with a sugar content of 12°BX according to the inoculum size of 5%, and cultured at 28° C. and 120 rpm for 72 hours to obtain a mature fermentation broth. After testing, the concentration of 4-vinylguaiacol and ethyl acetat...

Embodiment 3

[0035] The liquid state fermentation of embodiment 3 Pichia anomala GJYL08 strain

[0036] Fermentation was carried out in the same manner as in Example 2, except that step 3) was cultivated at 28° C. and 120 rpm for 120 h to obtain a mature fermentation broth. After testing, the concentration of 4-vinylguaiacol and ethyl acetate in the fermented broth reached 16.48 mg / L and 4.2 g / L.

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Abstract

A pichia anomala strain producing 4-ethenyl-2-methoxyphenol and ethyl acetate with high yields and applications thereof are disclosed. The pichia anomala strain is classified and named as pichia anomala GJYL08, and is deposited in the China Center for Type Culture Collection on March 13th, 2015, and the accession number of the pichia anomala is CCTCC NO:M2015109. The pichia anomala can grow in a large scale in a sorghum saccharified liquid medium. 12.48 mg / L of the 4-ethenyl-2-methoxyphenol and 3.6 g / L of the ethyl acetate can be obtained after the pichia anomala is subjected to shaking culture at 28 DEG C for 72 h with a speed being 120 rpm.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Pichia anomaly strain with high yield of 4-vinylguaiacol and ethyl acetate. Background technique [0002] 4-vinyl guaiacol is also known as vinyl guaiacol, 2-methoxy-4-vinylphenol, p-vinyl guaiacol, and its molecular formula is C 9 h 10 o 2 , molecular weight 150Da, colorless or light yellow oily liquid, fermented aroma, slightly sweet, slightly phenolic smell, is a main aroma component that determines the taste and quality of liquor, wine, soy sauce, tea, coffee, cheese and other foods At the same time, it is also one of the rare high-end spices in the daily chemical, pharmaceutical, flavor synthesis and other industries. At present, the chemical synthesis method is a common method for producing 4-vinyl guaiacol. As people's requirements for green products are increasing day by day, the method of producing 4-vinyl guaiacol by using biotechnology has the advantages o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16C12P7/22C12P7/62C12R1/84
Inventor 周庆伍李红歌汤有宏刘国英何宏魁李晓欢李安军高江婧
Owner ANHUI RUISIWEIER TECH
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