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Saccharomyces cerevisiae strain with multiple-stress resistance, and application thereof in cellulose alcohol fermentation

A technology for Saccharomyces cerevisiae and alcohol fermentation, which is applied in the directions of fermentation, fungi, microorganism-based methods, etc., can solve the problems of affecting the active ethanol yield of cells, inactive and unusable yeast cells, etc., and achieves good industrial application prospects, good Effect of inhibitor tolerance, low ethanol high yield

Inactive Publication Date: 2011-09-28
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Description
  • Claims
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Problems solved by technology

However, there are still some urgent problems to be solved in the application of Saccharomyces cerevisiae to ferment lignocellulose hydrolyzate to produce ethanol. At present, the process of using cellulase to degrade lignocellulose into fermentable sugar is usually at a relatively high temperature (about 50°C). However, the currently used yeast has a relatively good fermentation activity at 30°C. Under high temperature conditions, the yeast cells have no activity or even die, so it takes a certain period of cooling before the fermentation of the yeast can be carried out; lignocellulose is degraded to During the process of fermentable sugar and microbial fermentation, many substances that inhibit microbial growth and fermentation will be produced, such as acetic acid, ethanol, furfural, etc. Saccharomyces cerevisiae cannot use the five-carbon sugar in the cellulose hydrolyzate, which seriously affects cell activity and ethanol yield

Method used

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  • Saccharomyces cerevisiae strain with multiple-stress resistance, and application thereof in cellulose alcohol fermentation
  • Saccharomyces cerevisiae strain with multiple-stress resistance, and application thereof in cellulose alcohol fermentation
  • Saccharomyces cerevisiae strain with multiple-stress resistance, and application thereof in cellulose alcohol fermentation

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preparation example Construction

[0029] The preparation method of corn stalk hydrolyzate: Coarsely pulverize corn stalk (dry material), steam explosion for 5 minutes under 1.8MPa pressure (each process 5kg dry material); the material after steam explosion treatment is added into water, make the quality of dry material 100% The corn stalk content was 30%, cellulase (Shandong Zesheng Biotechnology Co., Ltd.) was added, and the amount of cellulase added was 20 FPU / g dry material, and enzymatic treatment was performed at 50° C. to obtain corn stalk hydrolyzate.

[0030] The detection method of total reducing sugar content is as follows:

[0031] (1) Preparation of solution

[0032] Dissolve 1 gram of dinitrosalicylic acid in 20ml of 2M aqueous sodium hydroxide solution and 50ml of water, then add 30 grams of potassium sodium tartrate tetrahydrate, add water to make up to 100ml, and store in a light-proof and airtight container, which is the DNS solution.

[0033] (2) Drawing of standard curve

[0034] Prepare 1...

Embodiment 1

[0044] Embodiment 1, the acquisition of Saccharomyces cerevisiae (Saccharomyces cerevisiae) T43

[0045] 1. Ion beam mutagenesis of Saccharomyces cerevisiae strain R8-10-2

[0046] Saccharomyces cerevisiae R8-10-2 has been preserved in the General Microorganism Center of China Committee for Culture Collection of Microorganisms (CGMCC for short, address: Datun Road, Chaoyang District, Beijing, China) on August 14, 2009, and the preservation number is CGMCC №. 3226 (see patent CN101633896; 200910091841.7). Saccharomyces cerevisiae R8-10-2 (CGMCC No.3226) is a yeast strain that can tolerate high concentrations of acetic acid. The strain can resist 0.8% acetic acid (volume percentage) at 30°C and 0.6% acetic acid ( Volume percentage), 40 ℃ can resist 0.2% acetic acid, the highest growth temperature is 42 ℃.

[0047] Saccharomyces cerevisiae R8-10-2 was cultured in the yeast complete medium YPD at 30°C until the mid-logarithmic phase (about 16 hours), centrifuged at 3000×g for 5 ...

Embodiment 2

[0063] Embodiment 2, Saccharomyces cerevisiae T43 and Saccharomyces cerevisiae R8-10-2 fermentability comparison at different temperatures

[0064] Saccharomyces cerevisiae T43 and Saccharomyces cerevisiae R8-10-2 were subjected to the following experiments simultaneously (the radius of rotation of the shaker was 25 mm):

[0065] 1. Preparation of seed solution

[0066] Saccharomyces cerevisiae (T43 or R8-10-2) was inoculated in YPD medium, cultured on a shaker at 30°C (200rpm) for 18 hours, transferred to 50ml of YPD medium according to the inoculation amount of 10% (volume percentage), and shaken at 30°C The bed (200rpm) was cultivated for 20 hours, and the obtained bacterial solution was the seed solution (OD600 value was about 6.0).

[0067] 2. Fermentation to produce ethanol

[0068] The seed liquid that step one obtains is connected in the shaking flask of the 250ml that 100ml ethanol fermentation medium EFM is housed by the inoculum size of 10% (percentage by volume),...

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Abstract

The invention discloses a saccharomyces cerevisiae strain with multiple-stress resistance, and the application of the saccharomyces cerevisiae strain in cellulose alcohol fermentation. The invention provides a saccharomyces cerevisiae T43 CGMCC No. 4642. The saccharomyces cerevisiae T43 has strong temperature adaptability (such that cooling and heating cost is reduced during fermentation processes) and good tolerance with inhibitors in cellulose hydrolysate (such that production processes can be simplified, and fermentation time can be shortened). Therefore, the saccharomyces cerevisiae has good prospect to be adopted in industrial application. According to the present invention, the efficient conversion of glucose in corn stalk hydrolysate to ethanol can be achieved, and the saccharomyces cerevisiae can be directly used in present fuel ethanol production technologies and new lignocellulose ethanol technologies. With the technical scheme of the present invention, ethanol production theorugh cerevisiae fermentation of cellulose hydrolysate can be started at a high temperature. According to the present invention, low energy consumption, low material consumption, low cost and high ethanol yield can be achieved in the cerevisiae fermentation process of cellulose alcohol production. With the present invention, huge economic benefits and good social benefits are brought in.

Description

technical field [0001] The invention relates to a Saccharomyces cerevisiae strain with multi-stress resistance, in particular to the application of the strain in ethanol fermentation, especially cellulose ethanol fermentation. Background technique [0002] At present, due to the rapid increase in the use of fossil energy such as oil, natural gas and coal, energy shortage has become a worldwide problem. The development of energy diversification and the acceleration of renewable energy development have become the research and development priorities of countries all over the world. The diversification strategy of energy supply represented by alternative energy such as fuel ethanol has become an important direction of my country's energy policy. In recent years, the production of fuel ethanol from starch or sugar has developed rapidly, which has alleviated the energy shortage to a certain extent, but has caused food security problems. Lignocellulose is a very abundant and cheap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/18C12P7/08C12R1/865
Inventor 何秀萍程艳飞郭雪娜卢莹张博润
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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