Journal
FUNGAL BIOLOGY
Volume 120, Issue 5, Pages 679-689Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.funbio.2016.03.002
Keywords
Antarctic fungi; Biosynthesis; Cold-active SOD; Cu/Zn-SOD gene sequence; Enzyme characterization
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Funding
- National Scientific Fund of the Ministry of Education and Science, Bulgaria [DO02-172/08]
- Fonds de la Recherche Fondamentale et Collective
- Belgian Program of Interuniversity Attraction Poles [iPros P7/44]
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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 degrees C caused 1.4-fold increase of specific SOD activity compared to unstressed culture. Maximum enzyme productivity was 64 x 10(3) 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. (C) 2016 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.
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