Production, purification, and characterization of a novel cold-active superoxide dismutase from the Antarctic strain Aspergillus glaucus 363
Autor: | Radoslav Abrashev, Maria Gerginova, Ekaterina Krumova, Jeni Miteva-Staleva, Nedelina Kostadinova, Boryana Spasova, Georges Feller, Spassen V. Vassilev, Zlatka Alexieva, Maria Angelova |
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Rok vydání: | 2016 |
Předmět: |
0106 biological sciences
0301 basic medicine Aspergillus glaucus Antarctic Regions Biology Mass spectrometry 01 natural sciences Mass Spectrometry Superoxide dismutase 03 medical and health sciences 010608 biotechnology Genetics Peptide sequence Ecology Evolution Behavior and Systematics Conserved Sequence chemistry.chemical_classification Molecular mass Strain (chemistry) Sequence Homology Amino Acid Superoxide Dismutase Sequence Analysis DNA Cold Temperature Molecular Weight Zinc 030104 developmental biology Infectious Diseases Enzyme Aspergillus Biochemistry chemistry biology.protein Dismutase Copper |
Zdroj: | Fungal biology. 120(5) |
ISSN: | 1878-6146 |
Popis: | The Antarctic fungal strain Aspergillus glaucus 363 produces cold-active (CA) Cu/Zn-superoxide dismutase (SOD). The strain contains at least one gene encoding Cu/Zn-SOD that exhibited high homology with the corresponding gene of other Aspergillus species. To our knowledge, this is the first nucleotide sequence of a CA Cu/Zn-SOD gene in fungi. An effective laboratory technology for A. glaucus SOD production in 3 L bioreactors was developed on the basis of transient cold-shock treatment. The temperature downshift to 10 °C caused 1.4-fold increase of specific SOD activity compared to unstressed culture. Maximum enzyme productivity was 64 × 103 U kg−1 h−1. Two SOD isoenzymes (Cu/Zn-SODI and Cu/Zn-SODII) were purified to electrophoretic homogeneity. The specific activity of the major isoenzyme, Cu/Zn-SODII, after Q-Sepharose chromatography was 4000 U mg−1. The molecular mass of SODI (38 159 Da) and of SODII (15 835 Da) was determined by electrospray quadropole time-of-flight (ESI-Q-TOF) mass spectrometry and dynamic light scattering (DLS). The presence of Cu and Zn were confirmed by inductively coupled plasma mass spectrometry (ICP-MS). The N-terminal amino acid sequence of Cu/Zn-SODII revealed a high degree of structural homology with Cu/Zn-SOD from other fungi, including Aspergillus species. |
Databáze: | OpenAIRE |
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