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Method for producing 4-guaethol through microbial fermentation

A technology for ethyl guaiacol and microbial fermentation, which is applied in the field of bioengineering, can solve the problems of low 4-EG content, low related enzyme activity, can not meet industrial production and the like, and achieves simple production process, wide sources, and fermentation performance. excellent effect

Inactive Publication Date: 2014-05-21
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many microorganisms can use the nutrients in the medium to produce 4-ethylguaiacol, but due to the low activity of related enzymes in the cells of these microorganisms, the content of 4-EG produced is low (generally 2.0-5.0mg / L ), so it cannot meet the requirements of industrial production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A method utilizing microbial fermentation to produce 4-ethylguaiacol, comprising the steps of:

[0037] (1) Inoculate the liquid seeds of the Bacillus circulans HG12 strain at a volume ratio of 5% into 200 mL of liquid fermentation medium, and ferment for 60 h at 40 °C and 200 r / min to obtain a fermentation broth. The fermented liquid was filtered and sterilized with a microporous membrane with a pore size of 0.2 μm under a pressure of 0.1 MPa, the filtrate was collected, and the pH value was adjusted to 5.0 to obtain a filtrate containing 4-vinylguaiacol.

[0038] (2) Inoculate the liquid seeds of Brettanomyces into the filtrate obtained in step (1) with an inoculum volume ratio of 10%, and ferment for 36 hours at 30°C to obtain 4-ethyl guaiac phenolic fermentation broth.

[0039] (3) Filter and sterilize the fermented liquid prepared in step (2) with a microporous membrane with a pore size of 0.6 μm under a pressure of 0.05 MPa, collect the filtrate, add butyl acetat...

Embodiment 2

[0043] The method utilizing microbial fermentation to produce 4-ethylguaiacol as described in Example 1, the difference is:

[0044] In step (1), the liquid seeds of Bacillus circulans HG12 strain were inoculated into 200 mL of liquid fermentation medium at an inoculation volume of 3% by volume, fermented for 36 hours at 37°C and 160 r / min, and filtered out The pH of the liquid is 6.0.

[0045] In step (2), liquid seeds of Brettanomyces were inoculated into the filtrate obtained in step (1) at an inoculum volume of 5%, and fermented at 28° C. for 12 hours.

[0046] In step (3), butyl acetate is added to the obtained filtrate, and the amount of butyl acetate is 20% (v / v) of the volume percentage of the filtrate.

[0047] The content of 4-ethylguaiacol in the fermentation broth was detected to be 1.01g / L.

[0048] Under these conditions, 0.202 g of 4-ethylguaiacol was obtained.

Embodiment 3

[0050] The method utilizing microbial fermentation to produce 4-ethylguaiacol as described in Example 1, the difference is:

[0051] In step (2), the Brettanomyces strain is Brettanomyces sp. CICC31575 (preserved by China Industrial Microorganism Culture Collection and Management Center).

[0052] The content of 4-ethylguaiacol in the fermentation broth was detected to be 1.15g / L.

[0053] Under these conditions, 0.230 g of 4-ethylguaiacol was obtained.

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Abstract

The invention belongs to the technical field of bioengineering and particularly relates to a method for producing 4-guaethol through microbial fermentation. A bacillus circulans HG12 strain is used as a production strain in the method. The method is characterized by comprising the steps of inoculating a liquid seed of the HG12 strain into a liquid fermentation medium to ferment to obtain a fermentation solution, filtering the fermentation solution and collecting filtrate; inoculating a liquid seed of a brettanomyces strain into the filtrate to ferment to obtain a 4-guaethol contained fermentation solution; filtering the fermentation solution, collecting the filtrate, and adding an extracting agent into the filtrate to obtain a 4-guaethol contained extracting solution; carrying out vacuum evaporation on the extracting solution to obtain the 4-guaethol. The method can be used for continuously producing 4-guaethol in a large scale; moreover, 4-guaethol is simple and convenient in production process, short in production period, low in production cost and extremely wide in industrial application prospect.

Description

(1) Technical field [0001] The invention belongs to the technical field of bioengineering, in particular to a method for producing 4-ethylguaiacol by microbial fermentation. (2) Background technology [0002] 4-Ethylguaiacol (4-Ethylguaiacol, 4-EG for short), also known as 2-methoxy-4-ethylphenol, has the molecular formula C 9 h 12 o 2 , with a molecular weight of 152.19Da and a density of 1.067×10 3 kg / m 3 , with a melting point of 15°C and a boiling point of 234-236°C. 4-EG is a colorless to light yellow oily liquid with a warm spicy taste. It belongs to the sauce-flavored spice and is the main flavor component of soy sauce and an important flavor substance of the sauce-flavored liquor. It not only endows the product with a specific It has unique style and taste, and has the function of scavenging free radicals, which can resist aging and prevent the occurrence of many diseases. At present, 4-ethylguaiacol has been widely used in soy sauce, wine, beverage, food, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/22C12R1/09C12R1/645
Inventor 董永胜马蕾崔云前段元良
Owner QILU UNIV OF TECH
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