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.
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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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