三氟乙醇和乙腈诱导纤维素酶形成熔融球蛋白状态和聚集体。
Trifluoroethanol and acetonitrile induced formation of the molten globule states and aggregates of cellulase.
机构信息
Department of Biochemistry, Faculty of Life Science, Aligarh Muslim University, Aligarh, India.
出版信息
Int J Biol Macromol. 2012 May 1;50(4):932-8. doi: 10.1016/j.ijbiomac.2012.02.012.
A systematic investigation on the effects of trifluoroethanol and acetonitrile at various concentrations on cellulase (EC 3.2.1.4) was studied by enzyme assay, intrinsic fluorescence, ANS binding, circular dichroism and ATR-Fourier transform infra red spectroscopy. The results show the presence of molten globule states at 3% (v/v) TFE and 80% (v/v) ACN. Cellulase aggregates at 25% (v/v) TFE and 90% (v/v) ACN, as detected by decrease in intrinsic and ANS fluorescence, loss in tertiary structure and enzyme activity, increase non-native β-sheet structure as evident from far-UV CD and FTIR spectra, increase in Thioflavin T fluorescence and shift in Congo red assay.
通过酶活性测定、内源荧光、ANS 结合、圆二色性和 ATR-FTIR 光谱法，系统研究了不同浓度的三氟乙醇和乙腈对纤维素酶（EC 3.2.1.4）的影响。结果表明，在 3%（v/v）TFE 和 80%（v/v）ACN 存在无定形球蛋白状态。在 25%（v/v）TFE 和 90%（v/v）ACN 时，纤维素酶聚集，如内源和 ANS 荧光强度降低、三级结构和酶活性丧失、远紫外 CD 和 FTIR 光谱显示非天然β-折叠结构增加、硫代黄素 T 荧光增加和刚果红试验中出现位移等所证实。
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