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Vanillin-tolerant saccharomyces cerevisiae

A technology of Saccharomyces cerevisiae and vanillin, which is applied in microorganism-based methods, fungi, microorganisms, etc., can solve the problems of incomplete elucidation of the inhibition mechanism of phenolic compounds, lack of quantitative and qualitative analysis methods, etc.

Inactive Publication Date: 2010-12-29
SHANDONG UNIV
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Larsson et al. obtained recombinant bacteria with improved coniferaldehyde tolerance by expressing the white-rot fungus Trametes versicolor laccase gene in yeast, but improved tolerance to certain phenolic compounds by domesticating and screening in lignocellulosic raw material hydrolyzate The strains have not been reported, and due to the lack of quantitative and qualitative analysis methods, the inhibitory mechanism of phenolic compounds on eukaryotic microorganisms such as Saccharomyces cerevisiae has not been fully elucidated

Method used

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  • Vanillin-tolerant saccharomyces cerevisiae
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  • Vanillin-tolerant saccharomyces cerevisiae

Examples

Experimental program
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Effect test

Embodiment 1

[0017] Embodiment 1 Saccharomyces cerevisiae directed domestication

[0018] The starting strain used for directional domestication is Saccharomyces cerevisiae EM-13, and the domestication steps are as follows:

[0019] (1) After activation, Saccharomyces cerevisiae strain EM-13 was first cultured in YEPD liquid medium (20%-SECE-YEPD) prepared with a concentration of 20% corn stalk steam explosion material extract (hereinafter referred to as SECE), and inoculated Amount is OD 初始 =0.1, when the bacteria grow to the logarithmic phase, the same inoculum size (OD 初始 =0.1) were transferred to the same medium (20%-SECE-YEPD), and the transfer was repeated in this way, and each transfer was called an acclimatization batch.

[0020] (2) During the acclimatization process, monitor the OD value and glucose consumption of each batch at 11 hours. When the glucose consumption at 11 hours no longer increases and is basically stable, increase the SECE concentration of the next batch of acc...

Embodiment 2

[0025] Embodiment 2 domesticated culture screening

[0026] The domesticated culture obtained in Example 1 was pre-screened, pre-screened and re-screened to finally obtain vanillin-tolerant Saccharomyces cerevisiae, and the strain was named EMV-8 (preserved in "China Microorganisms" on July 16, 2010 Species Preservation Management Committee General Microbiology Center", the preservation number is CGMCC No.4012), and the screening steps are as follows:

[0027] (1) Pre-screening on the dilution plate: Dilute the acclimatization culture solution to a suitable concentration and spread it on the pre-screening solid medium plate, and also dilute the culture solution of strains NAN-127 and EM-13 to a suitable concentration and spread it on the pre-screening As a control, culture on a solid medium plate in an incubator at 30°C. During the culture, observe the colony growth time, colony size and color, and select the colony with a shorter growth time, large colony and normal color (se...

Embodiment 3

[0035] Example 3 Detection of vanillin tolerance of bacterial strain EMV-8

[0036] The bacterial strain EMV-8 obtained by screening in Example 2 is tested for vanillin tolerance, and the steps are as follows:

[0037] (1) Activation of slant strains: respectively pick 1 ring of the slant-preserved strain EMV-8 and control strains EM-13 and NAN-27 and culture them in 20mL YEPD liquid medium. The shaker speed is 200r / min, preferably at 30°C Oxygen culture for 24 hours.

[0038] (2) Seed preparation: OD according to inoculum size 初始 =0.2 The cultures in step (1) were respectively transferred to 20 mL YEPD liquid medium for culture, the speed of the shaker was 200 r / min, and aerobic culture was carried out at 30° C. for 12 hours.

[0039] (3) Shake flask culture: OD according to the inoculum size 初始 =0.1 The cultures in step (2) were respectively transferred to the vanillin tolerance detection medium for culture, the shaker speed was 200r / min, and the oxygen-limited culture wa...

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Abstract

The invention discloses vanillin-tolerant saccharomyces cerevisiae. The strain is named as saccharomyces cerevisiae EMV-8 and collected in CGMCC on July 16th, 2010 with a collection number CGMCC No.4012. The strain is obtained by domesticating saccharomyces cerevisiae EM-13 in a YEPD liquid culture medium prepared from corn straw steam blackout extraction solution of different concentrations and screening; compared with the original strain EM-13, the strain can consume all glucose faster and has high ethanol yield when cultured in the YEPD liquid culture medium containing 1g / L of vanillin,; when cultured in the YEPD liquid culture medium containing 2g / L of vanillin, the strain has the growth delay period shortened by about 20 hours compared with the original strain; and the strain has good application value for further breeding the inhibitor tolerant saccharomyces cerevisiae in lignocellulose hydrolysis solution, and meanwhile has certain theoretical value for researching the phenolic compound tolerance mechanism of the saccharomyces cerevisiae.

Description

technical field [0001] The invention relates to a strain of Saccharomyces cerevisiae, in particular to a strain of Saccharomyces cerevisiae tolerant to vanillin and detection of its tolerance to vanillin, belonging to the field of microbial breeding. Background technique [0002] At present, the production of ethanol by fermentation is mainly based on starchy raw materials, the production cost is high, and there is a problem of competing with others for food and land. From the perspective of developing renewable clean energy and protecting the ecological environment, the research on the production of ethanol by using abundant and cheap renewable lignocellulosic raw materials has gradually attracted people's attention. Pretreatment is a necessary link in the production of ethanol from lignocellulose. This process will produce a variety of compounds that inhibit the growth of Saccharomyces cerevisiae, a microorganism that enzymatically hydrolyzes cellulose and subsequent ethan...

Claims

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Application Information

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IPC IPC(8): C12N1/18C12R1/865
Inventor 鲍晓明
Owner SHANDONG UNIV
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