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A use of proline for enhancing bacterial strain tolerance, a use of genes, an expression vector, a use of the expression vector, a bacterial strain, and a use of the bacterial strain

An expression vector, proline technology, applied in the biological field, can solve the problems of strain molecular modification restrictions, no reports, few target genes, etc.

Active Publication Date: 2016-05-18
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Most of the resistant strains reported in the literature are directed against a single inhibitor, and there are few reported target genes that can directly regulate the resistance of strains to inhibitors, especially for genes related to compound inhibitor resistance. , causing restrictions to the subsequent molecular transformation of strains, and becoming a bottleneck for the industrialization of cellulosic ethanol

Method used

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  • A use of proline for enhancing bacterial strain tolerance, a use of genes, an expression vector, a use of the expression vector, a bacterial strain, and a use of the bacterial strain
  • A use of proline for enhancing bacterial strain tolerance, a use of genes, an expression vector, a use of the expression vector, a bacterial strain, and a use of the bacterial strain
  • A use of proline for enhancing bacterial strain tolerance, a use of genes, an expression vector, a use of the expression vector, a bacterial strain, and a use of the bacterial strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 The knockout of proline synthesis-related genes reduces the tolerance of bacterial strains to compound inhibitors

[0049] (1) Preparation of culture medium

[0050] Seed liquid medium: glucose 20g / L, peptone 20g / L, yeast powder 10g / L, sterilized at 121°C for 20min.

[0051] Seed solid medium: glucose 20g / L, peptone 20g / L, yeast powder 10g / L, yeast powder 20g / L, sterilized at 121°C for 20min.

[0052]Fermentation medium: glucose 20g / L, peptone 20g / L, yeast powder 10g / L, sterilized at 121°C for 20min, adding cellulose hydrolyzate inhibitors furfural, phenol and acetic acid before inoculation, so that the content of furfural was 1.04g / L L, the content of acetic acid is 4.24g / L, and the content of phenol is 0.4g / L.

[0053] (2) fermentation

[0054] The strain BY4742 and the gene deletion strain BY4742-13659 (PRO1Δ), BY4742-11620 (PRO2Δ), BY4742-12141 (CAR1Δ), BY4742-16042 (CAR2Δ) were activated on the seed solid medium containing 200 μg / mL, and then picked A...

Embodiment 2

[0057] Example 2 Overexpression of the key gene PRO1 in the proline synthesis pathway enhances the tolerance of the strain to compound inhibitors

[0058] (1) Preparation of culture medium

[0059] SC-Ura liquid medium: glucose 20g / L, YNB6.7g / L, amino acid default mixture 2g / L, 20mg / L histidine, 20mg / L leucine, 20mg / L tryptophan, extinguished at 115℃ bacteria for 15 minutes.

[0060] SC-Ura solid medium: glucose 20g / L, YNB6.7g / L, amino acid default mixture 2g / L, 20mg / L histidine, 20mg / L leucine, 20mg / L tryptophan, 20g / L Agar powder, sterilized at 115°C for 15 minutes.

[0061] Fermentation medium: SC-Ura liquid medium, add cellulose hydrolyzate inhibitor furfural, phenol and acetic acid before inoculation, make the content of furfural be 0.78g / L, the content of acetic acid be 3.18g / L, the content of phenol is 0.3g / L.

[0062] (2) Plasmid construction and yeast transformation

[0063] Using the genome of Saccharomyces cerevisiae S288C as a template, using HXT7p_F in Table ...

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Abstract

The invention relates to the technical field of biology and particularly relates to a use of proline for enhancing bacterial strain tolerance to a composite inhibitor, a use of genes, an expression vector, a use of the expression vector, a bacterial strain, and a use of the bacterial strain. Tolerance of the bacterial strain to the composite inhibitor comprising furfural, acetic acid and phenol is enhanced.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of proline-enhanced bacterial strains tolerance to compound inhibitors, the application of genes, expression vectors and applications thereof, bacterial strains and applications thereof. Background technique [0002] In recent years, international conflicts arising from oil and other energy issues have intensified, and have gradually become major issues that plague the economic and social development of countries around the world. In order to cope with the increasingly serious energy crisis and get rid of the heavy dependence on imported oil, the development of new alternative energy has become a focus of attention of scholars from all over the world. Renewable, abundant, and cheap lignocellulose such as crop straw is used as raw material to produce Fuel ethanol is one of the most important directions. [0003] The production process of cellulosic ethanol mainly incl...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12N15/81C12P7/06C12R1/865
CPCY02E50/10
Inventor 元英进王昕李炳志白雪
Owner TIANJIN UNIV