High ferulic acid stress resistance clostridium beijerinckii and application thereof
A technology of Clostridium beijerinckii and ferulic acid, which is applied in the direction of bacteria, microorganisms, biochemical equipment and methods, can solve the problems of production stoppage of bio-butanol manufacturers, and achieve high butanol production, strong stress resistance, and repeated good sex effect
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Embodiment 1
[0031] This example illustrates the first step of plasma mutagenesis of the original strain of Clostridium beijingii.
[0032] The method for the first step plasma mutagenesis of the original strain of Clostridium beijerinckii is as follows:
[0033] Activate the original strain of Clostridium beijerinckii NCIMB8052 at a culture temperature of 33~37℃, a 50ml Schott anaerobic bottle with a volume of 15~20ml, nitrogen gas for 3min, and a culture time of 12~18h to obtain a vigorous bacterial solution; take fresh Cultured cells are diluted to cell concentration OD 600 =1~1.5, drip on the sterilized and cooled slide, dry with sterile air; use helium as the discharge gas, 100W as the radio frequency power, 10SLM as the gas flow, and 10~1800s as the irradiation time Plasma mutagenesis was performed on the strain, and after the mutagenesis, the bacterial membrane on the carrier was eluted and the survival rate was calculated. The experimental results are attached figure 1 Shown by figure...
Embodiment 2
[0035] This example illustrates the method of screening excellent Clostridium beijerinckii.
[0036] Among them, the medium formula used (% is the mass percentage):
[0037] (1) Solid plate medium: yeast powder 0.3%, peptone 0.5%, soluble starch 1%, ammonium acetate 0.2%, sodium chloride 0.2%, magnesium sulfate heptahydrate 0.3%, potassium dihydrogen phosphate 0.1%, dihydrogen phosphate Potassium 0.1%, ferrous sulfate heptahydrate 0.01%, agar 1.5%, pH 6.
[0038] (2) Resazurin plate medium: yeast powder 0.3%, peptone 0.5%, soluble starch 1%, ammonium acetate 0.2%, sodium chloride 0.2%, magnesium sulfate heptahydrate 0.3%, potassium dihydrogen phosphate 0.1%, phosphoric acid Dipotassium hydrogen 0.1%, ferrous sulfate heptahydrate 0.01%, agar 1.5%, resazurin 0.002%, ferulic acid 0.5%, pH 6.
[0039] (3) Seed culture medium: yeast powder 0.3%, peptone 0.5%, soluble starch 1%, ammonium acetate 0.2%, sodium chloride 0.2%, magnesium sulfate heptahydrate 0.3%, potassium dihydrogen phosphate...
Embodiment 3
[0050] This example illustrates the passage stability of the mutant M11.
[0051] In the fermentation medium using glucose as the carbon source, the passage stability of the mutant strain M11 was tested. The results of the passage fermentation test of the strain M11 are shown in Table 2:
[0052] Table 2
[0053]
[0054] From the experimental results, it can be seen that after 7 consecutive passages, the total solvent yield and butanol yield of the two mutant strains are relatively stable, with good passage stability, and can be used as production strains for further research and development.
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