Furfural-tolerant actinobacillus succinogenes GXAS-137FM as well as breeding method and application thereof
A technology of GXAS-137FM and Actinobacillus, applied in the direction of using microorganisms, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problems of unfavorable industrial production, polluting the environment, cumbersome operation, etc., and achieve economic benefits and Significant social benefits, reduced production costs, and the effect of solving environmental pollution problems
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Embodiment 1
[0059] Inoculate a single colony of Actinobacillus succinogenes GXAS137 into the seed medium, and culture it in an anaerobic incubator at 37°C for 18 hours. The number of bacteria reaches 350 million, and the secondary expansion culture is carried out according to the inoculation amount of 5% (V / V). 8h, when the number of bacteria reaches 300 million, the initial bacterial strain of the inoculation is obtained; the initial bacterial strain is inoculated to contain 0g / L, 0.2g / L, 0.4g / L, 0.6g / L, 0.8g / L, 1.0g / L furfural fermentation medium. The pH of the fermentation medium was adjusted to 6.8 by basic magnesium carbonate, and the fermentation was carried out at 36° C. for 72 hours at a rotation speed of 110 r / min. Samples were taken at 16h, 18h, 22h, 38h, 44h, 60h and 72h, respectively, and the biomass of GXAS137 bacteria and the concentration of succinic acid in the fermentation medium with different furfural concentrations were measured. For the results of bacterial biomass,...
Embodiment 2
[0063] A single colony of Actinobacillus succinogenes GXAS137 was inserted into the seed medium and cultured in an anaerobic incubator at 36°C for 18 hours, and the number of bacteria reached 360 million. According to 8% (V / V) inoculum size, transfer to the fermented medium containing furfural and carry out acclimatization, when growing to the stationary phase, according to the fermented medium containing furfural 1.0 g / L, 1.2g / L, 1.4g successively / L, 1.6g / L concentrations were increased for the next round of acclimatization, and the experiment was repeated to obtain a better furfural tolerance strain Actinobacillus succinogenes GXAS-137FM.
Embodiment 3
[0065] Insert the GXAS-137FM obtained in Example 2 into the seed culture medium, cultivate in an anaerobic incubator at 38°C for 20 hours, and the number of bacteria reaches 400 million, and then carry out secondary expansion and cultivation according to the inoculation amount of 5% (V / V) After 10 hours, when the number of bacteria reaches 340 million, inoculate respectively in the furfural fermentation medium that adds 1.0g / L, 1.2g / L, 1.4g / L and 1.6g / L according to the inoculation amount of 12% (V / V). Fermentation, the fermentation conditions are shown in Example 1, and the fermentation time is 48h. After the fermentation is finished, the biomass of the bacteria in the fermentation medium and the content of succinic acid are measured according to the assay method of Example 1. see results image 3 .
[0066] Depend on image 3 It can be seen that the GXAS-137FM strain grows under the fermentation medium of 1.2g / L, 1.4g / L and 1.6g / L furfural concentration. The output of dia...
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