Method for vapor-exploding stalk enzymolysis coupling ferment for hydrogen production by using immobilized cell
A technology for immobilizing cells and steam-exploding straws is applied in the field of fermentation technology, which can solve the problems of difficulty in improving fermentation efficiency, difficulty in normal cellulase activity, etc., and achieves the effects of high resistance, easy separation, and release of product inhibition.
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Embodiment 1
[0031] Embodiment 1. Using the method provided by the present invention to use steam-exploded corn stalks as raw materials to carry out enzymatic hydrolysis coupled fermentation hydrogen production, the specific steps are as follows:
[0032] 1. Put the raw material of steam-exploded corn stalks in an enzymolysis tank, add 25 IU / g-substrate of cellulase and tap water with pH 4.8 adjusted with sulfuric acid, the water contains urea 2g / l, KH 2 PO 4 0.51g / l, MgSO 4 0.31g / l, the solid-to-liquid ratio of steam-exploded straw and aqueous solution is 1:8 (w / v), enzymatically hydrolyzed at 50°C;
[0033] 2. Preparation of Clostridium butyricum (C.butylicum AS1.209) seed culture:
[0034] Each liter of seed medium includes the following components: glucose 20g, yeast extract 3g, KH 2 PO 4 0.2g, K 2 HPO 4 1.6g, MgSO 4 ·7H 2 O0.2g, NaCl0.1g, CaCl 2 0.01g, Na 2 S·9H 2 O0.25g, NaMoO 4 2H 2 O0.01g, NaHCO 3 0.2g and (NH 4 ) 2 SO 4 3.0 g; Clostridium butyricum was inoculated ...
Embodiment 2
[0038] Embodiment 2. Using the method provided by the present invention to use steam-exploded wheat straw as raw material to carry out enzymatic hydrolysis coupling fermentation hydrogen production, the specific steps are as follows:
[0039] 1. Put the raw material of steam-exploded wheat straw in an enzymolysis tank, add 25IU / g-substrate of cellulase and tap water with pH 4.8 adjusted with sulfuric acid, the water contains 2g / l of yeast extract, KH 2 PO 4 0.51g / l, MgSO 4 0.31g / l, the solid-to-liquid ratio of steam-exploded straw and aqueous solution is 1:10 (w / v), enzymatically hydrolyzed at 50°C;
[0040] Step 2 is the same as embodiment 1;
[0041] 3. Mix the seed solution of Clostridium butyricum (C.butyricum AS1.209) in the logarithmic growth phase with a sterile 3% sodium alginate solution, and the volume ratio of the seed solution to the sodium alginate solution is 1:4. Use a sterile syringe to draw the mixture, slowly drop the mixture into sterile 0.05mol / l CaCl 2...
Embodiment 3
[0044] Example 3. Using the method provided by the present invention to use steam-exploded rice straw as raw material to carry out enzymatic hydrolysis coupled fermentation hydrogen production, the specific steps are as follows:
[0045] 1. Put the raw material of steam-exploded rice straw in an enzymolysis tank, add 25 IU / g-substrate of cellulase and tap water with pH 4.8 adjusted with sulfuric acid, the water contains urea 2g / l, KH 2 PO 4 0.51g / l, MgSO 4 0.31g / l, the solid-to-liquid ratio of steam-exploded straw and aqueous solution is 1:12 (w / v), enzymatically hydrolyzed at 50°C.
[0046] Step 2 is the same as in Example 1.
[0047] 3. Mix the seed solution of Clostridium butyricum (C.butyricum AS1.209) in the logarithmic growth phase with a sterile 4% sodium alginate solution, and the volume ratio of the seed solution to the sodium alginate solution is 1:5. Use a sterile syringe to draw the mixture, slowly drop the mixture into sterile 0.05mol / l CaCl 2 solution, the mi...
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