High yield recombinant production of a self-assembling polycationic peptide for silica biomineralization
| Autor: | Stephan Hobe, Sandor Nietzsche, Christian Zerfaß, Sandra Braukmann, Harald Paulsen |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
chemistry.chemical_classification
Diatoms Oligopeptide Ion chromatography Peptide Isomerase medicine.disease_cause Silicon Dioxide Recombinant Proteins law.invention chemistry.chemical_compound chemistry Biochemistry law medicine Recombinant DNA Escherichia coli Cyanogen bromide Oligopeptides Biotechnology Biomineralization |
| Zdroj: | Protein expression and purification. 108 |
| ISSN: | 1096-0279 |
| Popis: | We report the recombinant bacterial expression and purification at high yields of a polycationic oligopeptide, P5S3. The sequence of P5S3 was inspired by a diatom silaffin, a silica precipitating peptide. Like its native model, P5S3 exhibits silica biomineralizing activity, but furthermore has unusual self-assembling properties. P5S3 is efficiently expressed in Escherichia coli as fusion with ketosteroid isomerase (KSI), which causes deposition in inclusion bodies. After breaking the fusion by cyanogen bromide reaction, P5S3 was purified by cation exchange chromatography, taking advantage of the exceptionally high content of basic amino acids. The numerous cationic charges do not prevent, but may even promote counterion-independent self-assembly which in turn leads to silica precipitation. Enzymatic phosphorylation, a common modification in native silica biomineralizing peptides, can be used to modify the precipitation activity. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect