A mutant strain of Zymomonas mobilis tolerant to low pH and its application

A technology of Zymomonas and mutant strains is applied in the field of microorganisms to achieve the effects of improving fermentation efficiency, reducing production costs and reducing autoclaving links

Active Publication Date: 2020-12-04
BIOGAS SCI RES INST MIN OF AGRI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, this patent creates a mutant strain that can normally ferment and produce ethanol in the low pH environment of pH = 4 and pH = 3.5. This process does not require high temperature and high pressure sterilization, and there is no other report in this species that has reached this level.

Method used

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  • A mutant strain of Zymomonas mobilis tolerant to low pH and its application

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Experimental program
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Effect test

Embodiment 1

[0028] Screening of embodiment 1 mutant strain PH1-29

[0029] 1) Irradiation mutagenesis was carried out on Zymomonas mobilis cells by normal temperature plasma mutagenesis breeding technology. First, the starting strain ZM4 was activated and cultivated at a temperature of 30°C for 16 hours;

[0030] 2) Then take the overnight culture (10 6 -10 8 cells), centrifuge at 4°C for 10 min at 3000 rpm, wash the bacteria with normal saline and resuspend in 1 mL of normal saline; take 10 μL of the resuspended cells and place them in an ARTP mutagenesis breeding instrument for 15-120 s;

[0031] 3) Prepare solid medium: 50g / L glucose, 2g / L potassium dihydrogen phosphate, 10g / L yeast extract, and use sulfuric acid solution to adjust the pH of the medium;

[0032] 4) Resuspending the cells after irradiation mutagenesis for 16 hours, and then spreading them on a solid medium with pH=4 for culture;

[0033] 5) All colonies were selected and screened on a medium with pH=3.5, and the scre...

Embodiment 2

[0034] Embodiment 2 common fermentation culture medium prepares ethanol

[0035] 1) Preparation of fermentation medium: 50g / L glucose, 2g / L potassium dihydrogen phosphate, 10g / L yeast extract, and use sulfuric acid solution to adjust the pH of the medium to 4;

[0036] 2) inoculate the mutant strain PH1-29 and the starting strain ZM4 on the prepared medium respectively, the inoculum amount is 10%, and carry out fermentation and cultivation for 40 h;

[0037] 3) ethanol is separated and the conversion rate is measured.

Embodiment 3

[0038] Embodiment 3 common fermentation culture medium prepares ethanol

[0039] 1) Preparation of fermentation medium: 50g / L glucose, 2g / L potassium dihydrogen phosphate, 10g / L yeast extract, adjust medium pH=3.5 with sulfuric acid solution

[0040] 2) inoculate the mutant strain PH1-29 and the starting strain ZM4 on the prepared medium respectively, the inoculation amount is 10%, and carry out fermentation and cultivation for 90 h;

[0041] 3) ethanol is separated and the conversion rate is measured.

[0042] Preparation of ethanol from common fermentation medium at normal pH in comparative example

[0043] 1) Preparation of fermentation medium: 50g / L glucose, 2g / L potassium dihydrogen phosphate, 10g / L yeast extract, adjust the pH of the medium to 6.6 with sulfuric acid solution, and sterilize the medium at 115°C for 20 minutes;

[0044] 2) Inoculate the mutant strain PH1-29 and the starting strain ZM4 on the prepared medium respectively, the inoculum amount is 10%, and ca...

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Abstract

The invention discloses a mutant strain of Zymomonas mobilis tolerant to low pH value and its application. The strain is named PH1-29 in taxonomy, and the preservation number is GDMCC 60260; obtained by repeated selection on low pH medium. The strain can not only ensure normal growth in a low pH environment and ensure good fermentation efficiency, but also can eliminate pollution due to the low pH environment, and can also avoid high-temperature and high-pressure sterilization in the fermentation process, control production costs, and improve fermentation efficiency.

Description

technical field [0001] The invention belongs to the field of microbes, and in particular relates to a mutant strain of Zymomonas mobilis tolerant to low pH and its application, which can be applied to the production of fuel ethanol bio-based products in any open fermentation environment without high pressure sterilization. Background technique [0002] In the modern bio-fermentation industry, high-temperature sterilization is an important and crucial link, which is related to fermentation costs and product safety, so it deserves attention. Most microorganisms do not grow well in a low pH environment, so lowering the pH is a strategy to reduce the risk of bacterial contamination. However, the pH tolerance of most fermentation strains is not high. If the mutation method is used to screen the fermentation strains that are tolerant to the low pH environment, not only can the fermentation efficiency be ensured, but also the low pH environment can eliminate pollution and avoid hi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P7/06C12R1/01
CPCC12N1/20C12P7/06C12N1/205C12R2001/01Y02E50/10
Inventor 秦晗吴波何明雄
Owner BIOGAS SCI RES INST MIN OF AGRI
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