2-phenylethanol-producing yeast strain, its culturing method and application

A culture method and strain technology, applied in the field of microorganisms, can solve the problem of high synthesis cost of 2-phenylethanol, achieve high tolerance and increase yield

Inactive Publication Date: 2012-12-12
QILU UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Cui Zhifeng (Cui Zhifeng, Che Zhibo, Yang Xiao, etc. Breeding of 2-phenylethanol-tolerant high-yielding yeast strains [J]. Journal of Zhejiang University of Technology, 2008 (36): 427-430.) and Etschmann M (Etschmann M, Bluemkee W, Sell D, et al. Biotechnological production of 2-phenylethanol [J]. Appl Microbiol Biotechnol, 2002, 59:1-8.) respectively reported the method of single-phase batch fermentation of yeast strains to produce 2-phenylethanol , but on the whole, the synthesis cost of 2-phenylethanol is still high, and the yeast strains obtained from the above research results are still difficult to meet the needs of the market, so finding higher-yielding 2-phenylethanol microbial strains is still the key to solving the above problems

Method used

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  • 2-phenylethanol-producing yeast strain, its culturing method and application

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Embodiment 1

[0043] A strain of Saccharomyces cerevisiae (Saccharomyces cerevisiae) WHH6 strain was preserved in the General Microbiology Center of China Committee for the Collection of Microbial Cultures on December 20, 2010, address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, The deposit number is CGMCC No.4491.

[0044] Saccharomyces cerevisiae (Saccharomyces cerevisiae) WHH6 strain has typical characteristics of Saccharomyces cerevisiae, the colony is white, with rounded edges and a slightly raised center, such as figure 1 As shown; the cells are nearly round, with an average size of 15-21 μm×14-16 μm (length×width), such as figure 2 Shown; Fermentation produces a strong aroma, and the ethanol production can reach 9.8% (v / v). The single-phase single strain can ferment and produce 2-phenylethanol up to 5.4003g / L in the optimized medium (Table 1). It can be used as 2-phenylethanol, wine and feed yeast productio...

Embodiment 2

[0046] The culture method of Saccharomyces cerevisiae (Saccharomyces cerevisiae) WHH6 bacterial strain, the steps are as follows:

[0047] (1) Pick 1-2 rings of Saccharomyces cerevisiae WHH6 from the inclined surface of the test tube and insert it into the liquid seed medium, and activate and shake it for 18-22 hours under the conditions of 28-30°C and 120-160r / min to obtain activated liquid seeds ;

[0048] (2) Inoculate the liquid seeds prepared in step (1) into the liquid fermentation medium at an inoculum size of 8-12% (v / v), and ferment and cultivate at 28-30°C and 120-160r / min. After culturing for 24 h, samples were taken every 12 h to determine the OD of the fermentation broth 660 value and 2-phenylethanol content in the supernatant (results such as Figure 5 shown), after 40 h of fermentation culture, the content of 2-phenylethanol in the fermentation supernatant and the OD of the bacterial liquid were measured every 4 h 660 value (results such as Figure 6 shown),...

Embodiment 3

[0085] The application of Saccharomyces cerevisiae (Saccharomyces cerevisiae) WHH6 strain in the preparation of 2-phenylethanol, the steps are as follows:

[0086] (1) Pick 2-ring Saccharomyces cerevisiae WHH6 from the slant of the test tube and insert it into the liquid seed medium, and activate and shake it for 20 hours at 28°C and 120r / min to obtain activated liquid seeds;

[0087] (2) Inoculate the liquid seeds prepared in step (1) into the liquid fermentation medium at an inoculum size of 8% (v / v), and culture and ferment them at 28°C and 120r / m for 24 to 40 hours respectively. Add 10% sterilized oleic acid in the volume of the liquid fermentation medium at the time point, continue to cultivate at 28°C and 120r / min until the total time is 56h, centrifuge at 6000r / min for 10min, take the oil phase, and obtain 2- phenethyl alcohol.

[0088] Preferably according to the present invention, the components of the liquid seed medium in the step (1) are as follows:

[0089] Yeas...

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Abstract

The invention relates to a 2-phenylethanol-producing yeast strain, its culturing method and application. The Saccharomyces cerevisiae WHH6 strain provided in the invention is preserved on December 20, 2010 in the General microbiological center of China Committee for Culture Collection of Microorganisms. The preservation address is: Institute of Microbiology of Chinese Academy of Sciences, No.3, No.1 yard, Beichen West Road, Chaoyang District, Beijing. The spawn preservation number is CGMCC No.4491. The Saccharomyces cerevisiae WHH6 strain screened out by the invention has the characteristic of high 2-phenylethanol yield, which is detected to be able to reach 5.8564g / L. And the strain has high tolerance to 2-phenylethanol, thus being in favor of application of the strain in the industrial production of 2-phenylethanol.

Description

technical field [0001] The invention relates to a 2-phenylethanol-producing yeast strain and a culture method and application thereof, belonging to the technical field of microorganisms. Background technique [0002] 2-Phenylethyl alcohol, also known as β-phenylethyl alcohol, is an aromatic alcohol with a rose aroma. It naturally exists in various plant leaves, flowers and extracted essential oils. It is abundant in fermented foods such as bread, fruit wine, cheese and soy sauce. . 2-Phenylethyl alcohol is the main component of rose fragrance, and it is also a supplementary component of various fragrance types. Its derivative esters such as phenylethyl acetate are also important aromatic products. At present, the global production of 2-phenylethanol exceeds 10,000 tons, most of which are chemically synthesized, and only a small amount is extracted from plant essential oils. The cost of the chemical synthesis method is relatively low, but the raw material benzene or styrene...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/18C12P7/22C12R1/865
Inventor 刘天明唐育岐刘欣欣
Owner QILU UNIV OF TECH
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