Escherichia coli LL016 and application thereof
A technology of Escherichia coli and succinic acid, applied in the direction of bacteria, microorganisms, biochemical equipment and methods, etc.
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Embodiment 1
[0037] This example illustrates the method of performing the first step of plasma mutagenesis on the original strain of Escherichia coli.
[0038] The method for carrying out the first step plasma mutagenesis of Escherichia coli original strain is as follows:
[0039] Activate the original strain of Escherichia coli BA016 in an LB test tube, culture overnight at 37°C, 200 r / min; take freshly cultured cells and dilute to the cell concentration OD 600 =1.0, drop on the sterile slide, and dry with sterile wind; use helium as the discharge gas, 80-120 W as the radio frequency power, 10-30 SLM as the gas flow rate, and 10-30 s as the radiation According to the time, plasma mutagenesis was performed on the strain; after mutagenesis, the bacterial film on the slide was eluted with 1 mL of sterile saline, spread on a synthetic medium plate, and cultivated in an anaerobic box. The experimental results are attached figure 1 , where the abscissa is the irradiation time, and the ordina...
Embodiment 2
[0041] This example illustrates the method for screening excellent Escherichia coli.
[0042] Wherein, the medium formula used is as follows:
[0043] (1) Solid synthetic medium plate: citric acid 3 g L -1 , Na 2 HPO 4 12H 2 O 4 g L -1 , KH 2 PO 4 8 g L -1 , (NH 4 ) 2 HPO 4 8 g L -1 , NH 4 Cl 0.2 g L -1 , (NH 4 ) 2 SO 4 0.75 g·L -1 , MgSO 4 ·7H 2 O 1 g L -1 , CaCl 2 2H 2 O 10.0 mg·L -1 , ZnSO 4 ·7H 2 O 0.5 mg L-1 , CuCl 2 2H 2 O 0.25 mg L -1 , MnSO 4 ·H 2 O 2.5 mg L -1 , CoCl 2 ·6H 2 O 1.75 mg L -1 , H 3 BO 3 0.12 mg·L -1 , Al 2 (SO 4 ) 3 1.77 mg·L -1 , Na 2 MoO 4 2H 2 O 0.5 mg L -1 , ferric citrate 16.1 mg·L -1 , Vitamin B 1 20 mg·L -1 , biotin 2 mg·L -1 , agar 20 g / L, glucose 10 g / L.
[0044] (2) Seed medium: peptone 10 g / L, yeast powder 5 g / L, NaCl 5 g / L.
[0045] (3) Shake flask fermentation medium: citric acid 3 g L -1 , Na 2 HPO 4 12H 2 O 4 g L -1 , KH 2 PO 4 8 g L -1 , (NH 4 ) 2 HPO 4 8 g L -1 , NH 4...
Embodiment 3
[0057] This example illustrates the passage stability of the mutant strain LL016
[0058] On the synthetic medium plate, the mutant strain LL016 was transferred and cultured several times, and the obtained strains were subjected to fermentation verification respectively. The experimental results are shown in Table 2:
[0059] Table 2 Passage stability analysis of mutant strain LL016
[0060]
[0061] It can be seen from the experimental results that after 8 consecutive subcultures, the succinic acid production and succinic acid conversion rate of the mutant strain LL016 are relatively stable, with good passage stability, and can be used as a production strain for further research and development.
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