Cloning, overexpression, crystallization and preliminary X-ray analysis of a family 1 β-glucosidase fromStreptomyces
| Autor: | Miquel Vallmitjana, Rosa Perez, Enrique Querol, Alicia Guasch, Miquel Coll, Josep A. Pérez-Pons |
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| Rok vydání: | 1999 |
| Předmět: |
Ammonium sulfate
Protein Conformation Crystallography X-Ray medicine.disease_cause Streptomyces law.invention Hydrolysis chemistry.chemical_compound Structural Biology law Escherichia coli medicine Glycoside hydrolase Cloning Molecular Crystallization DNA Primers Base Sequence biology Beta-glucosidase beta-Glucosidase General Medicine biology.organism_classification Recombinant Proteins Crystallography chemistry Orthorhombic crystal system |
| Zdroj: | Acta Crystallographica Section D Biological Crystallography. 55:679-682 |
| ISSN: | 0907-4449 |
| DOI: | 10.1107/s0907444998013833 |
| Popis: | An intracellular beta-glucosidase (Bgl3) from Streptomyces sp. has been cloned and overexpressed in Escherichia coli. The introduction of a His tag at the N-terminal end of the protein has allowed its purification to homogeneity by a single chromatographic step, with yields of 150-200 mg of pure protein per litre of E. coli culture. The enzyme (52.6 kDa) is a retaining glycosidase able to hydrolyze a wide range of disaccharides and oligosaccharides and to perform transglycosylation. Crystals of recombinant Bgl3 have been grown from an ammonium sulfate solution using the hanging-drop vapour-diffusion method at 293 K. The crystals belong to the orthorhombic space group I222 with unit-cell dimensions a = 101.6, b = 113.4 and c = 187.5 A at room temperature and contain two molecules per asymmetric unit. A full 1.69 A resolution diffraction data set (97.7% completeness) has been collected from frozen crystals in a solution containing 30% sucrose, using synchrotron radiation. |
| Databáze: | OpenAIRE |
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