Fernandes Sheryl, Mattiasson Bo, Hatti-Kaul Rajni
Department of Biotechnology, Center for Chemistry & Chemical Engineering, Lund University, P.O. Box 124, S-221 00 Lund, Sweden.
Biotechnol Prog. 2002 Jan-Feb;18(1):116-23. doi: 10.1021/bp010161m.
Foam generated by vigorous stirring of a nonionic detergent, Triton X-114, was used for the recovery of recombinant cutinase expressed by Saccharomyces cerevisiae. The enzyme with a hydrophobic fusion tag, (Trp-Pro)(4), was recovered with a higher yield as compared to the wild-type cutinase, indicating the involvement of hydrophobic interactions in protein isolation with the foam. The influence of various factors including volume, dilution, pH, different additives, and cell concentration in the medium on enzyme recovery was investigated. Interaction of the enzyme with detergent was monitored using fluorescence spectroscopy. No significant changes in protein conformation after the isolation procedure were observed using circular dichroism.
通过剧烈搅拌非离子洗涤剂Triton X-114产生的泡沫用于回收酿酒酵母表达的重组角质酶。与野生型角质酶相比，带有疏水融合标签(Trp-Pro)(4)的酶回收率更高，这表明疏水相互作用参与了用泡沫进行的蛋白质分离过程。研究了包括体积、稀释度、pH值、不同添加剂以及培养基中细胞浓度等各种因素对酶回收率的影响。使用荧光光谱监测酶与洗涤剂的相互作用。通过圆二色性观察到分离过程后蛋白质构象没有显著变化。
