Immobilising proteins on silica using modified silaffin peptides
نویسندگان
چکیده
1.1 Materials ........................................................................................................................ 2 1.2 Instrumentation .............................................................................................................. 2 1.3 HPLC and mass spectrometry ........................................................................................ 3 1.4 Cloning of Thioredoxin (TRX)-GyrA-H6-CBD fusion protein..................................... 3 1.5 Protein expression and purification ............................................................................... 4 1.6 DTT-mediated hydrolysis of TRX-MESNa thioester .................................................... 4 1.7 Scanning electron microscopy and fluorescence microscopy ........................................ 5 1.8 Quantification of precipitated silica ............................................................................... 5 1.9 Stability of silica immobilised eGFP against denaturation with SDS ............................ 5 1.10 Quantification of loading of silica with TRX and release of TRX from silica .............. 5 1.11 Determination of enzyme activity of TRX and silica immobilised TRX ...................... 6 2 Supplementary data ................................................................................................................ 7
منابع مشابه
Immobilising proteins on silica with site-specifically attached modified silaffin peptides.
Immobilisation of proteins on solid supports such as silica is commonly applied to improve performance of enzymes under detrimental conditions and to allow enzyme recycling. Silica biomineralisation processes occurring in nature have recently inspired approaches towards mild, biomimetic silica formation. In diatoms, complex posttranslationally modified silaffin peptides are directly involved in...
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