Method for producing L-lactic acid through continuous fermentation
A technology of lactic acid and lactic acid bacteria, applied in the field of fermentation, can solve the problem of high cost of lactic acid fermentation, achieve the effects of reducing strain preparation, reducing production cost, and improving fermentation intensity
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Embodiment 1
[0028] Example 1 Preparation of crop waste saccharification solution
[0029] Crush the corncob, add it to the reactor, then add 2% (w / v) NaOH at a solid-to-liquid ratio of 1:5 (w / w), seal the mouth, and heat it at 125°C for 50 Minutes, the obtained slurry is centrifuged in a 2000rpm centrifuge, and the centrifuged black liquor is treated with a membrane to recover the alkali therein; the obtained solid phase is washed with water and added to the container, and 2 % (v / v) dilute HCl, adjust the temperature to 50°C for 1 hour, then add cellulase according to 20 units / g of cellulose, stir at 100 rpm, and control the enzymolysis time at 65 hours. After the end of enzymolysis, the material The liquid is centrifugally filtered, and the liquid phase is enzymatic sugar solution. The total sugar concentration of the enzymatic solution is measured by DNS method, and the concentration of glucose and xylose in the enzymatic solution is measured by high performance liquid chromatography. ...
Embodiment 2
[0030] Example 2 The first generation of L-lactic acid continuous fermentation
[0031] Taking Lactobacillus 1.2131 (purchased from the Institute of Microbiology, Chinese Academy of Sciences) as an example, the fermentation process of the present invention will be described.
[0032] For the preserved lactic acid bacteria, first prepare the seed liquid through the following process:
[0033] Plate culture: Inoculate the lactic acid bacteria into the activated medium and culture at 30°C for 40 hours;
[0034] Shake flask seed culture: Inoculate the lactic acid bacteria cultured on the plate into the seed liquid medium, culture anaerobically at 38°C for 20 hours, shake the table at a speed of 200rpm, and detect the OD 600 is 4.0.
[0035] Wherein, the saccharification solution prepared in Example 1 is used as the carbon source, and the mass percentage of the activated medium used for plate culture is prepared as follows: 1% total sugar, 0.5% yeast extract, 0.5% peptone, and 2%...
Embodiment 3
[0040] Example 3 The second generation of L-lactic acid continuous fermentation
[0041]Use the lactic acid bacteria after 10 batches of fermentation in Example 2 to inoculate into the fermentation medium, wherein, using the saccharification solution prepared in Example 1 as the carbon source, the mass percentage of the second-generation continuous fermentation medium is configured as follows: total sugar 15 %, Yeast Extract 0.2%~0.4%, Beef Extract 0.3%~0.5%, Potassium Dihydrogen Phosphate 0.03%~0.05%, Dipotassium Hydrogen Phosphate 0.03%~0.05%, Sodium Chloride 0.02%~0.03%, Magnesium Sulfate 0.04% %~0.06%, calcium carbonate 3%~5%, carry out anaerobic stirring at 40°C and 200rpm, and continue to ferment; ferment until the lactic acid content no longer increases, stop the fermentation, stop stirring, release 5 / 6 volume of fermentation broth, and replenish Add new fermentation medium to continue fermentation; 10 batches of continuous cycle fermentation, the fermentation period is...
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