Expression, purification and preliminary crystallographic studies of alpha-amylase isozyme 1 from barley seeds
| Autor: | Xavier Robert, Richard Haser, Tine E. Gottschalk, Birte Svensson, Nushin Aghajari |
|---|---|
| Přispěvatelé: | Deleage, Gilbert, Institut de biologie et chimie des protéines [Lyon] (IBCP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2002 |
| Předmět: |
Polyethylene glycol
Biology Crystallography X-Ray Isozyme Pichia Pichia pastoris chemistry.chemical_compound Structural Biology [SDV.BBM] Life Sciences [q-bio]/Biochemistry Molecular Biology [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Amylase chemistry.chemical_classification food and beverages Embryo Hordeum General Medicine biology.organism_classification Recombinant Proteins Isoenzymes Enzyme chemistry Biochemistry Germination Seeds biology.protein Orthorhombic crystal system alpha-Amylases |
| Zdroj: | Acta Crystallogr D Biol Crystallogr Acta Crystallogr D Biol Crystallogr, 2002, 58, pp.683-686 Technical University of Denmark Orbit HAL |
| Popis: | International audience; The germinating barley seed contains two major alpha-amylase isozyme families, AMY1 and AMY2, involved in starch degradation to provide energy used by the plant embryo for growth. Many years of difficulty in growing three-dimensional crystals of natural AMY1 have now been overcome by a nonapeptide truncation of the enzyme C-terminus. The truncated enzyme was overexpressed in Pichia pastoris, purified and crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol 8000 as precipitant and 2-propanol as an additive. Crystals belong to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 88.36, b = 72.82, c = 61.74 A and one molecule per asymmetric unit.The germinating barley seed contains two major alpha-amylase isozyme families, AMY1 and AMY2, involved in starch degradation to provide energy used by the plant embryo for growth. Many years of difficulty in growing three-dimensional crystals of natural AMY1 have now been overcome by a nonapeptide truncation of the enzyme C-terminus. The truncated enzyme was overexpressed in Pichia pastoris, purified and crystallized by the hanging-drop vapour-diffusion method using polyethylene glycol 8000 as precipitant and 2-propanol as an additive. Crystals belong to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 88.36, b = 72.82, c = 61.74 A and one molecule per asymmetric unit. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|