Actinobacillus succinogenes strain and method for producing succinic acid by screening and fermentation of same
A technology for producing actinomycetes and succinic acid, which is applied in the field of bioengineering and can solve problems such as difficult to know technical conditions, low yield, and unreachable yield
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Embodiment 1
[0061] Screening and identification of Actinobacillus succinate strains:
[0062] 1. Preparation of enrichment medium: a medium containing fumarate as the sole carbon source and supplemented with nitrogen source, inorganic salts and monensin. The detailed formula is as follows (g / L): sodium fumarate 10-30, yeast powder 5-20, peptone 5-20, K 2 HPO 4 1-5, CaCl 2 0.1-1, NaCl 0.5-2, (NH 4 ) 2 SO 4 0.5-2, MgCl 2 0.1-1, monensin 0.1-0.2, the enrichment medium can be obtained by mixing the above-mentioned raw materials. It has been verified by experiments that the enrichment medium prepared by any concentration combination in the aforementioned enrichment medium raw material formula can screen out the GXAS137 strain of the present invention.
[0063] 2. Preparation of plate screening medium: add sodium succinate and agar powder on the basis of enrichment medium. The detailed formula is as follows (g / L): sodium fumarate 10-30, yeast powder 5-20, peptone 5-20, K2 HPO 4 1-...
Embodiment 2
[0112] According to the method for fermenting and producing succinic acid by the Actinobacillus succinic acid strain in Example 1, the plate of the bacterial strain with the preservation number CCTCC M 2011399 was cultured in an anaerobic box for 48 hours, and then connected to a 30 mL seed medium with an inoculation loop for 37 Cultivate at ℃ for 20 hours, insert the fermentation medium (250mL Erlenmeyer flask with a liquid volume of 150mL) of different carbon sources (65 g / L in terms of glucose content) according to the 5% inoculum amount, and culture statically for 48 hours. Diacid and main by-product content, the results are shown in Table 2.
[0113] carbon source Succinic acid content (g / L) Acetic acid content (g / L) Lactic acid content (g / L) glucose 47.98 10.68 1.63 sucrose 50.31 8.36 0.91 fructose 40.34 9.27 1.39 maltose 49.67 10.83 1.50 Arabic candy 54.76 7.24 1.37 Xylose 41.13 7.23 0 Mannitol 52.33 ...
Embodiment 3
[0116] According to the method for producing succinic acid by fermentation of the Actinobacillus succinigenes strain in Example 1, the plate of the strain with the preservation number CCTCC M 2011399 was cultured in an anaerobic box for 48 hours, and then connected to a 30 mL seed medium with an inoculation loop, at 37°C Cultivate for 20 hours, add different raw materials (75 g / L in terms of glucose content) to the fermentation medium (250 mL Erlenmeyer flask, liquid volume: 150 mL) according to 5% inoculum amount, and culture for 48 hours statically, and measure succinic acid by HPLC And the main by-product content, the results are shown in Table 3.
[0117] carbon source Succinic acid content (g / L) Acetic acid content (g / L) Lactic acid content (g / L) tapioca flour 70.17 10.77 1.46 corn flour 53.81 12.34 2.01 cane molasses 49.37 7.26 1.17 Sugar cane juice 58.67 8.91 1.64
[0118] It can be seen from Table 3 that when cassava ...
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