Method for preparing immobilized cellulase and method for hydrolyzing cellulose through immobilized cellulase
A technology of cellulase and cellulose, applied to biochemical equipment and methods, fixed on/in organic carriers, hydrolytic enzymes, etc., can solve problems such as increased steric hindrance, and achieve reduction of Michaelis constant, Improve the immobilization rate of enzyme activity and the effect of increasing the hole density
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[0024] Example 1
[0025] 1) Accurately weigh the cellulase powder and dissolve it in pure water to make the concentration 5mg / ml, and set aside. Cellulase is produced from Trichoderma viride.
[0026] 2) Preparation of immobilized cellulase: accurately weigh 2.625g polyethylene glycol with a molecular weight of 4000 into a 100ml beaker, add 48.25g 0.05mol / L citric acid-sodium citrate buffer solution with pH=4.8, room temperature After stirring and dissolving with a glass rod, 1.75 g of sodium alginate was added so that the mass fraction of sodium alginate was 3.5% and the mass ratio of sodium alginate to polyethylene glycol was 1:1.5. After mixing uniformly at room temperature, a sodium alginate-polyethylene glycol solution is formed, and it is set aside.
[0027] Use a 10ml syringe to suck 5ml of the above sodium alginate-polyethylene glycol solution into a 20ml glass bottle, add 1ml of 5mg / ml cellulase solution and 1.5ml of 1% glutaraldehyde solution in turn, and mix well, Shak...
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[0036] Example 2
[0037] The preparation of sodium alginate-polyethylene glycol immobilized cellulase, the operation is the same as that in Example 1(1)(2), the fixation conditions are respectively the lowest and highest values described in the "Summary of the Invention (1)" for each factor Optimize the conditions of the immobilized enzyme, and the results are shown in Table 3.
[0038] Sodium alginate-polyethylene glycol immobilized cellulase was used to hydrolyze microcrystalline cellulose, and the operation was the same as in Example 1(3)(4). The conditions of enzymatic hydrolysis are respectively taken as the lowest value and the highest value of each factor described in "Summary of the Invention (2)", and the conditions of the enzymatic hydrolysis reaction are optimized and analyzed, as shown in Table 4.
[0039] Table 3 Condition optimization analysis of immobilized cellulase
[0040]
[0041]
[0042] Table 4 Optimization analysis of enzymatic hydrolysis reaction conditions...
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