Strain and method for producing 2,3-butanediol and organic acid
An organic acid and butanediol technology, applied in the biological field, can solve the problems of sugar loss, increased process cost, and low tolerance of wild bacteria inhibitors
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Embodiment 1
[0064] Embodiment 1 The preparation of corncob hydrolyzate and screening medium
[0065] The corn cob was pulverized with a pulverizer, the solid particle size was 40-60 mesh, and the contents of the three main components of the corn cob were shown in Table 1. The corn cob was hydrolyzed with a mixed acid containing sulfuric acid and phosphoric acid, the mass fractions of sulfuric acid and phosphoric acid were 1.5% and 0.5% respectively. Weigh 20g of corncobs into a 500ml Erlenmeyer flask, add 60ml of mixed acid at a solid-to-liquid ratio of 1:3, seal it with tinfoil to prevent water loss, and treat it in a high-pressure steam sterilizer at 130°C for 60min. The filtrate was obtained by suction filtration, during which it was washed with 40 ml of hot distilled water (65° C.) in order to obtain more sugar. The pH of the filtrate obtained above was adjusted to 5.0 with sodium hydroxide, and the pH was further adjusted to 6.5 with ammonia water to obtain a corncob hydrolyzate. T...
Embodiment 2
[0071] Embodiment 2 plasma mutagenesis and screening
[0072] Use an inoculation loop to dip a small amount of bacterial liquid from the glycerol tube and streak it on the LB solid medium, culture it in a 30°C incubator for 14-16 hours, pick a single colony and transfer it to the LB liquid medium (50ml of liquid in a 250ml Erlenmeyer flask), Cultivate in a shaker at 30°C at 150rpm for 6-8 hours, take a sample every 1 hour to measure OD, take the bacterial solution in the logarithmic growth phase (OD≈1), use a pipette gun to take 10 μl of the bacterial solution and drop it on the center of a round stainless steel iron plate, and put the stainless steel The iron sheet is placed in the studio, with helium as the working gas, and the power supply is 120W. The plasma is treated for 0s, 60s, 120s, 180s, 240s, 300s, and 360s respectively. In a 1.5ml EP tube filled with 1ml LB medium, vibrate with a vortex shaker to fully elute the bacteria, and resuscitate on a shaker at 30°C at 150r...
Embodiment 3
[0075] Example 3 Production of 2,3-BDO and organic acids by fermentation of corncob hydrolyzate
[0076]Table 3 is the fermentation situation of the initial bacterial strain, which cannot grow in 100% corncob hydrolyzate, after the hydrolyzate is diluted to a certain number of times, the bacterial strain can grow, but the sugar concentration in the medium is diluted to some extent, resulting in a lower final yield.
[0077] Inoculate the 23 bacterial strains obtained by re-screening into the seed medium (250ml Erlenmeyer flask liquid volume 50ml), 30 ℃ shaking table 150rpm culture 14~16h, inoculate in the fermentation medium according to 10% inoculum amount (250ml Erlenmeyer flask liquid volume 50ml), fermented at 35°C in a shaker at 150rpm for 48h, and detected the content of each component by liquid chromatography. The 23 strains were stored in glycerol tubes as the first generation strains.
[0078] As shown in Table 4, when the 23 strains were fermented in 100% corn cob ...
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