A kind of preparation method of bacterial cellulose
A bacterial cellulose and seed technology, applied in the field of industrial biology, can solve problems such as immaturity, and achieve the effects of reducing fermentation costs, simple process, and low cost
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Embodiment 1
[0030] Example 1: Fermentation in 7L NBS tank
[0031] 1) Shake flask strain preparation: scrape a ring of inclined seeds and connect them to sterilized seed culture medium (glucose 20, yeast extract 2, peptone 2, CaCO3 0.5, unit g / L, natural pH value), 28°C, Cultivation at 100r / min for 16 hours followed by expansion;
[0032] 2) Preparation of molasses: pre-dilute the original sugar cane molasses by 2 times, adjust the pH to 2.0 with sulfuric acid, hydrolyze at 90°C for 30 minutes, leave it for one night, centrifuge or filter the supernatant and dilute it 10 times, pH 6.0;
[0033] 3) Fermentation: inoculate the seeds at 5% on a medium containing a certain amount of sugar cane molasses (molasses 25, glycerol 0.5, corn steep liquor 10g / L, ammonium sulfate 0.5, citric acid 1.0, disodium hydrogen phosphate 2.0, magnesium sulfate 0.2, Unit g / L) fermenter is stirred and fermented for 8 days; among them, the stirring tank is a bioreactor equipped with gauze, and the gauze is hung on the ...
Embodiment 2
[0035] Example 2: 50-liter stirred tank fermentation
[0036] 1) Strain preparation: scrape a ring of slant seeds and connect them to the sterilized seed medium (glucose 40, yeast extract 5, peptone 5, CaCO3 1, unit g / L, natural pH), 32℃, 150r / After 24 hours of culture in min, move to the NBS tank expansion in Example 1. The liquid volume is 3L, the culture medium is similar to the shake flask, the rotation speed is 100r / min, the aeration volume is 1.0vvm, and the temperature is 32°C for 20 hours.
[0037] 2) Preparation of molasses: pre-dilute the beet molasses by 5 times, adjust the pH to 2.5 with sulfuric acid, hydrolyze at 100°C for 20 minutes, let it stand overnight, centrifuge or filter the supernatant and dilute it by 2 times, the pH is 5.5;
[0038] 3) Fermentation: inoculate the seeds on a medium containing 30L molasses (molasses 50, glycerol 0.2, corn steep liquor 20g / L, ammonium sulfate 1.5, citric acid 1.5, disodium hydrogen phosphate 3.0, magnesium sulfate 0.6, unit g / L...
Embodiment 3
[0040] Example 3: Fermentation in a 2-ton stirred tank
[0041] 1) Strain preparation: scrape a ring of slant seeds and connect them to sterilized seed culture medium (glucose 25, yeast extract 2.5, peptone 2.5, CaCO 3 3. Unit g / L, natural pH value) 500mL Erlenmeyer flask, cultivated at 30°C and 150r / min for 20 hours, then use a 5L Erlenmeyer flask with 1L culture medium to cultivate for 20 hours under the same conditions. Move to a 200L seed tank containing 100L of culture medium. The culture medium is similar to a shaker bottle, with a rotation speed of 90r / min, aeration volume of 1.5vvm, and a temperature of 30°C for 24 hours with stirring;
[0042] 2) Preparation of molasses: Pre-dilute the sugar cane molasses by 4 times, adjust the pH to 3.0 with sulfuric acid, hydrolyze at 120°C for 15 minutes, leave it for one night, centrifuge or filter to take the supernatant and dilute it 2.5 times, the pH is 4.8;
[0043] 3) Fermentation: Transfer the seeds to a medium containing 1200L mo...
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