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Schizosaccharomyces pombe with high acetic acid tolerance

A technology of Schizosaccharomyces pombe and ethanol, applied in the field of microorganisms, can solve problems such as yeast fermentation inhibition, achieve ethanol resistance, good lactic acid resistance, and solve the effect of fermentation inhibition

Active Publication Date: 2021-05-25
贵州国台酒业集团股份有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to solve the above technical problems, the present invention provides a Schizosaccharomyces pombe with high acetic acid tolerance, which can normally ferment and produce ethanol in a medium with an acetic acid content of 20 g / L, and can solve the problem of yeast strains in high-concentration acetic acid environments. The problem of inhibited fermentation

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Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1 Molecular biology identification

[0030] 26SrDNA identification of the strain Schizosaccharomyces pombe Sujiu.014. The selected universal primers are:

[0031] NL1 (5'-GCATATCAATAAGCGGAGGAAAAG-3');

[0032] NL4 (5'-GGTCCGTGTTTCAAGACGG-3').

[0033] The length of the amplified fragment is about 600bp. The target fragment was sent to Jinweizhi for sequencing, and the sequencing results were compared with the 26S rDNA sequences registered in GenBank (KY296084.1, MT729825.1, NR_151433.1, CU329672.1, etc.), and the homology reached 98%, see Figure 6 . The strain was identified as Schizosaccharomyces pombe by 26S rDNA sequence analysis.

Embodiment 2

[0034] Example 2 Bacterial colony morphology, microscopic morphology observation

[0035] (1) Colony: the purified single colony was inoculated on WL medium by dilution and plating, cultured at 30°C for 2-3 days, and the colony morphology was observed and recorded. The result is as figure 1 As shown in -a, the colony is hemispherical, with neat edges, opaque, yellowish on the front and back, uniform texture and easy to stir up.

[0036] (2) Microscopic examination: Methylene blue staining, 40X oil microscope observation. The result is as figure 1 As shown in -b, the thallus is rod-shaped without flagella.

[0037] (3) Scanning electron microscope:

[0038] Culture the yeast on a shaker for 16 hours (stable period), take 5mL of the bacterial solution and add it to a 50mL centrifuge tube, centrifuge at 7000r / min for 2min, pour off the supernatant; add 20mL of 5% fixative glutaraldehyde solution, vortex and oscillate and let stand. Fix for 2 hours; add 20mL PBS buffer, wash ...

Embodiment 3

[0039] Embodiment 3 carbon and nitrogen source assimilation experiment

[0040] (1) Carbon source assimilation test

[0041] Assimilative carbon source basal medium: (NH 4 ) 2 SO 4 0.5%, KH 2 PO 4 0.1%, MgSO 4 ·7H 2 O 0.05%, yeast extract powder 0.02%, agar 2%, sterilized at 115°C for 15min.

[0042] The carbon sources tested were citric acid, galactose, xylose, ethanol, lactic acid, acetic acid, and arabinose, and they were sterilized by a filter membrane (0.20 μm) with a sterilized syringe to make the content reach 50 mmol / L.

[0043] Put the tested bacteria into 3mL sterile normal saline, shake well, draw 1mL of yeast suspension into a sterile petri dish, then pour into the carbon-source-free basal medium that has been melted and cooled to about 45-50°C, shake Mix well, and after solidification, place it at 30°C and incubate it upside down for 7 hours. Take out the bacteria-carrying plate, and divide it into six plots with a glass pencil on the bottom of the plate,...

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Abstract

The invention provides schizosaccharomyces pombe with high acetic acid tolerance, and belongs to the technical field of microorganisms. The invention relates to Schizosaccharomyces pombe Sujiu.014, in particular to Schizosaccharomyces pombe Sujiu.014, the strain is screened from Maotai-flavor liquor to produce fermented grains, and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.21791. The strain is schizosaccharomyces pombe which has extremely strong acetic acid tolerance and relatively good lactic acid tolerance, high temperature resistance and ethanol tolerance. The yeast strain can tolerate 20 g / L acetic acid, is a yeast strain with the highest acetic acid tolerance reported at present, and can tolerate 60 g / L lactic acid, 40 DEG C temperature and 8% ethanol at the same time. The problem that yeast fermentation is inhibited in a high-concentration acetic acid environment can be solved.

Description

Technical field: [0001] The invention provides a Schizosaccharomyces pombe with high tolerance to acetic acid, which belongs to the technical field of microorganisms. Background technique: [0002] Yeast is a multifunctional industrial microorganism that has made outstanding contributions in the fields of traditional fermented food and pharmaceutical production. At the same time, in the era of rapid development of the bioenergy industry, yeast is also a favorable conversion platform for environmentally friendly energy production due to its efficient biofuel production capacity. However, in the actual industrial production process, yeasts are often stressed by environmental factors such as extreme temperature, weak organic acid, and high osmotic pressure. All of these stressful conditions affect the growth rate and metabolic capacity of cells, thereby reducing productivity. Acetic acid is a typical weak organic acid that accumulates as a by-product or exists as an inhibitor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16C12G3/02C12P7/06C12R1/645
CPCC12G3/02C12P7/06Y02E50/10
Inventor 张翠英卢君肖冬光李长文林良才李超戎梓溢
Owner 贵州国台酒业集团股份有限公司
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