Acid Extraction and Purification of Recombinant Spider Silk Proteins

详细信息    查看全文

• 作者：Charlene M. Mello ; Jason W. Soares ; Steven Arcidiacono ; and Michelle M. Butler
• 刊名：Biomacromolecules
• 出版年：2004
• 出版时间：September 2004
• 年：2004
• 卷：5
• 期：5
• 页码：1849 - 1852
• 全文大小：64K
• 年卷期：v.5,no.5(September 2004)
• ISSN：1526-4602

文摘

A procedure has been developed for the isolation of recombinant spider silk proteins based upon their uniquestability and solubilization characteristics. Three recombinant silk proteins, (SpI)₇, NcDS, and [(SpI)₄/(SpII)₁]₄,were purified by extraction with organic acids followed by affinity or ion exchange chromatography resultingin 90-95% pure silk solutions. The protein yield of NcDS (15 mg/L culture) and (SpI)₇ (35 mg/L) increased4- and 5-fold, respectively, from previously reported values presumably due to a more complete solubilizationof the expressed recombinant protein. [(SpI)₄/(SpII)₁]₄, a hybrid protein based on the repeat sequences ofspidroin I and spidroin II, had a yield of 12.4 mg/L. This method is an effective, reproducible techniquethat has broad applicability for a variety of silk proteins as well as other acid stable biopolymers.