High-temperature resistant saccharomyces cerevisiae strain and breeding method thereof
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A Saccharomyces cerevisiae strain and technology of Saccharomyces cerevisiae, which are applied in the field of bioengineering, can solve problems such as weakening effects, and achieve the effects of strong high temperature tolerance, high genetic stability, and wide application prospects
Active Publication Date: 2013-08-07
贵州钓鱼台国宾酒业有限公司
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Breeding using traditional mutagenesis techniques, most yeast strains have a "passivation" response to mutagens, which greatly weakens the effect of traditional breeding techniques
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Embodiment 1
[0017] Embodiment 1: Breeding of high temperature resistant Saccharomyces cerevisiae strains
[0018] (1) Traditional mutagenesis breeding to establish a mutant strain library
[0019] Pick a ring of yeast slant culture and inoculate it in 5mL of YPD liquid medium, cultivate it at 30°C and 100rpm until the middle and late logarithmic growth period, collect the bacteria by centrifugation and wash twice with sterile saline, resuspend the cells to adjust the concentration to 10 6 individual / mL. Take 4mL of bacterial suspension in a Ф70mm petri dish, and irradiate it with a 15W ultraviolet lamp at a height of 30cm from the bacterial liquid in the petri dish. Then spread on the YPD medium plate. Irradiate the bacterial solution for different times (0s, 5s, 10s, 20s, 30s, 40s, 50s, 60s, 70s, 80s), respectively, dilute and plate for two days, wait for the colonies to grow out, count the colonies on the plate, and Calculate the lethality rate and draw the lethality rate curve base...
Embodiment 2
[0025] Embodiment 2: Fermentation performance of genome rearrangement bacterial strain
[0026] (1) High temperature tolerance of genome rearranged strains
[0027]The starting strain AY12 and the genome rearrangement strain LYQ-F3 were cultured to the logarithmic growth phase, respectively inoculated into Duchenne tubes containing wort medium (6°Brix), respectively, at set temperatures (30°C, 35°C, 38°C, 40°C, 42°C) for static culture, observe the gas production every 12 hours and record the results (as shown in Table 1). As shown in the results, after genome rearrangement, the gas production of the strain under high temperature conditions was significantly improved compared with the original strain, indicating that the high temperature tolerance of the genome rearranged strain was higher than that of the original strain.
[0028] Table 1 High temperature tolerance of starting strains and genome rearrangement strains
[0029]
[0030] Note: The data shown are representat...
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Abstract
The invention provides a high-temperature resistant saccharomyces cerevisiae strain and a breeding method of the high-temperature resistant saccharomyces cerevisiae strain. The saccharomyces cerevisiae (Saccharomyces cerevisiae) SY-5-11L has a preservation number of CGMCC No.7377. The strain is characterized in that the ethanol yield is 13.9% (v / v) at a condition of fermenting at high temperature of 38 DEG C on the premise that other fermenting performances are not influenced, the yield is improved by 17.8% as compared with that of a parent strain. The ethanol output at the condition of fermenting at high temperature of 40 DEG C is 12.2% (v / v), and is improved by 34.1% as compared with that of the parent strain. The breeding method comprises the following steps of: performing artificial ultraviolet mutagenesis on an initial strain of the saccharomyces cerevisiae; screening out 23 forward mutant strains as a genome rearrangement library, wherein the ethanol output of the forward mutant strains are obviously raised on the condition of fermenting at a high temperature; and performing the genome rearrangement at three rounds, thus gaining the mutant strains with the maximum ethanol output under the high-temperature fermenting condition. The finally bred saccharomyces cerevisiae strain can be subjected to alcoholic fermentation at the high-temperature fermenting condition, brings no special requirements to fermenting equipment and technological conditions, and can be put into service with the equipment and conditions of general plants, thus having a wide application prospect.
Description
Technical field: [0001] The invention belongs to the technical field of bioengineering, and relates to the breeding of industrial microorganisms, in particular to a high-temperature-resistant Saccharomyces cerevisiae strain and a breeding method thereof. Background technique: [0002] Alcohol occupies a very important position in the national economy and the people's livelihood, and is widely used in medicine, organic synthesis, food industry, industry and agriculture, and national defense industry. With the increasing demand for alcohol in recent years, the production of alcohol has attracted more and more attention. In 2001, the Chinese government included "vigorously develop fuel alcohol" in the "Tenth Five-Year Plan". In the demonstration of the "Tenth Five-Year Plan" development plan for vehicle fuel alcohol. [0003] At present, the raw materials for alcohol production are mainly starchy raw materials such as corn, molasses and fibrous raw materials, etc.; the strain...
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