Departamento Engenharia Biológica, Universidade do Minho, Campus de Gualtar 4710-057 Braga, Portugal.
Bioresour Technol. 2012 Apr;110:526-33. doi: 10.1016/j.biortech.2012.01.140. Epub 2012 Feb 6.
The recovery of cellulases from lignin, lignocellulosic hydrolysates and cellulose by alkaline washes at pH 9 and 10 was examined. The effect of the pH on the structural stability of purified Cel7A was analyzed by circular dichroism. Purified Cel7A showed conformational changes at pH 9 and 10 that were reversible at pH 4.8. Temperature influenced the enzymatic hydrolysis of wheat straw and may be critical for the efficiency of cellulase recycling from wheat straw hydrolysates. Operation at moderate temperatures (37°C) resulted in a rate of saccharification 19% higher than that obtained at 50° C, improving cellulase recycling by 49%. Over 60% of the enzyme activity on the synthetic substrate 4-methylumbelliferyl-β-d-cellobioside (MUC) may be recovered by using a simple alkaline wash. This is thus a promising strategy for enzyme recycling that is simple to implement at industrial scale, economical and effective.
考察了在 pH 值为 9 和 10 的碱性条件下从木质素、木质纤维素水解物和纤维素中回收纤维素酶的情况。通过圆二色性分析研究了 pH 值对纯化 Cel7A 结构稳定性的影响。在 pH 值为 9 和 10 时,纯化的 Cel7A 发生构象变化,在 pH 值为 4.8 时可恢复。温度影响小麦秸秆的酶解,可能对从小麦秸秆水解物中回收纤维素酶的效率至关重要。在中等温度(37°C)下操作比在 50°C 下的糖化速率高 19%,提高了 49%的纤维素酶回收效率。在合成底物 4-甲基伞形酮-β-D-纤维二糖苷(MUC)上可回收超过 60%的酶活性,使用简单的碱性洗涤即可实现。因此,这是一种很有前途的酶回收策略,在工业规模上简单易行、经济有效。