Method for saccharifying cellulose raw material
A technology for cellulose and cellulose degrading bacteria, applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of harsh requirements, polluted equipment, high energy consumption, etc., to avoid pollution and improve the degradation speed. , The effect of saving the cost of saccharification
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Embodiment 1
[0039] 1) Dry the corn stalks, crush them into 40-mesh particles, take 2g of crushed raw materials, add 100ml GS-2 liquid medium (1.5g potassium dihydrogen phosphate per liter, 2.9g dipotassium hydrogen phosphate, 10g MOPS, 3g dihydrogen phosphate Sodium citrate water, 2.1g urea, 1.0g magnesium chloride hexahydrate, 150mg calcium chloride dihydrate, 1.25mg ferrous sulfate hexahydrate, 6g yeast extract and 1g cysteine salt, and add 0.15ml 99% glacial acetic acid , pH=7.4), 0.2MPa nitrogen / hydrogen gas mixture (5% hydrogen) was passed through for 5 minutes to remove oxygen, and then sterilized at 121°C for 15 minutes;
[0040] 2) Prepare Clostridium thermocellum seed liquid, pick a single clone of Clostridium thermocellum, inoculate into 5ml GS-2 liquid medium (0.5% microcrystalline cellulose), and culture anaerobically at 60°C to OD 600 is 1.5;
[0041] 3) Inoculate 1ml of seed liquid into the medium containing corn stalks in step 1, and ferment in anaerobic sealed manner at...
Embodiment 2
[0044] 1) Prepare 100ml of GS-2 liquid medium containing 0.5% crystalline cellulose (1.5g potassium dihydrogen phosphate per liter, 2.9g dipotassium hydrogen phosphate, 10g MOPS, 3g sodium citrate dihydrate, 2.1g urea, 1.0g Magnesium chloride hexahydrate, 150mg calcium chloride dihydrate, 1.25mg ferrous sulfate hexahydrate, 6g yeast extract and 1g cysteine salt, pH=7.4), feed 0.2MPa nitrogen / hydrogen gas mixture (in which the hydrogen content is 5 %) deoxygenation for 5 minutes, 121 ° C sterilization for 15 minutes;
[0045] 2) Preparation of Clostridium thermocellum seed liquid: Pick a single clone of Clostridium thermocellum, inoculate it into 5ml GS-2 liquid medium (0.5% microcrystalline cellulose), and culture it anaerobically at 60°C for 36 hours to OD 600 is 1.5;
[0046] 3) Inoculate 1ml of seed solution into the culture medium of step 1, and ferment in anaerobic seal at 60°C for 72 hours, and then measure the filter paper enzyme activity of cellulosomes in the ferme...
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