Research Center in Applied Chemistry (CEPESQ), Department of Chemistry, Federal University of Paraná, P.O. Box 19081, Curitiba, Paraná 81531-970, Brazil; VTT Technical Research Centre of Finland, P.O. Box 1000, 02044 VTT Espoo, Finland.
Bioresour Technol. 2014 Jan;151:392-6. doi: 10.1016/j.biortech.2013.09.135. Epub 2013 Oct 7.
The activity profile of a 1:0.30 mixture of Celluclast 1.5L FG and Novozym 188 (Novozymes) was investigated using Whatman #1 filter paper (W1FP) as a single substrate for hydrolysis. The procedure was based on the ability of the enzymes to release total (RS(Tot)), insoluble (RS(Insol)) and soluble (RS(Sol)) reducing sugars from W1FP. RS(Insol) was used to estimate endoglucanase (EnG) activity whereas exoglucanases (ExG) were assessed by measuring RSSol in the presence of δ-gluconolactone. Finally, the β-glucosidase (βG) activity was derived from the difference between RS(Sol) measurements in the presence and absence of δ-gluconolactone. When this analytical procedure was applied to W1FP using 9.64 mg mL(-1) of the enzyme mixture, the relative contributions of EnG, ExG and βG to the total cellulase activity were 63.28%, 12.02% and 24.70%, respectively. Also, this ratio changed with changes in the enzyme loading, giving a new insight into the synergy that exists among the enzymes.
使用 Whatman #1 滤纸 (W1FP) 作为水解的单一底物，研究了 1:0.30 的 Celluclast 1.5L FG 和 Novozym 188（诺维信）混合物的活性谱。该过程基于酶从 W1FP 中释放总还原糖 (RS(Tot))、不溶性还原糖 (RS(Insol)) 和可溶性还原糖 (RS(Sol)) 的能力。RS(Insol) 用于估计内切葡聚糖酶 (EnG) 活性，而外切葡聚糖酶 (ExG) 通过测量 δ-葡萄糖酸内酯存在下的 RSSol 来评估。最后，β-葡萄糖苷酶 (βG) 活性来自于存在和不存在 δ-葡萄糖酸内酯时 RS(Sol) 测量值之间的差异。当将此分析程序应用于 W1FP 时，使用 9.64mg/mL(-1) 的酶混合物，EnG、ExG 和 βG 对总纤维素酶活性的相对贡献分别为 63.28%、12.02%和 24.70%。此外，该比例随酶加载量的变化而变化，这为酶之间存在的协同作用提供了新的见解。
