Effects of iron and manganese availability on growth and production of superoxide dismutase byStreptococcus suis

1999 ◽  
Vol 45 (12) ◽  
pp. 1027-1032 ◽  
Author(s):  
Donald F Niven ◽  
Andrew Ekins ◽  
Aws A-W Al-Samaurai
Keyword(s):  
Superoxide Dismutase ◽  
Manganese Superoxide Dismutase ◽  
Polyacrylamide Gel Electrophoresis ◽  
Streptococcus Suis ◽  
Complex Medium ◽  
Good Growth ◽  
High Concentration ◽  
Activity Staining ◽  
Cell Extracts ◽  
Manganese Availability

A complex medium supported good growth of the type strain of Streptococcus suis irrespective of the presence or absence of a high concentration (1 mM) of the iron chelating agent, ethylenediamine di-o-hydroxyphenylacetic acid. Good growth was also obtained using a complex medium that had been treated with Chelex-100 to reduce the iron content, but only if this medium was supplemented with manganese; supplementation with iron had little effect. Collectively, these results indicate that S. suis requires manganese, but not iron, for growth. Polyacrylamide gel electrophoresis of cell extracts followed by activity staining revealed the presence of a single manganese-cofactored superoxide dismutase; activity staining and enzyme assays revealed that manganese availability during growth affected the activity of the superoxide dismutase in cell extracts. The results are discussed with respect to the capacity of S. suis to avoid damage by reactive oxygen species.Key words: Streptococcus suis, iron, manganese, superoxide dismutase.

scholarly journals Molecular Characterization of a Recombinant Manganese Superoxide Dismutase fromLactococcus lactisM4

2014 ◽  
Vol 2014 ◽  
pp. 1-9
Author(s):  
Boon Hooi Tan ◽  
Thean Chor Leow ◽  
Hooi Ling Foo ◽  
Raha Abdul Rahim
Keyword(s):  
Superoxide Dismutase ◽  
Active Sites ◽  
Manganese Superoxide Dismutase ◽  
Specific Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Maximal Activity ◽  
Gel Filtration Chromatography ◽  
T7 Promoter

A superoxide dismutase (SOD) gene ofLactococcus lactisM4 was cloned and expressed in a prokaryotic system. Sequence analysis revealed an open reading frame of 621 bp which codes for 206 amino acid residues. Expression ofsodAunder T7 promoter exhibited a specific activity of 4967 U/mg when induced with 1 mM of isopropyl-β-D-thiogalactopyranoside. The recombinant SOD was purified to homogeneity by immobilised metal affinity chromatography and Superose 12 gel filtration chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot analyses of the recombinant SOD detected a molecular mass of approximately 27 kDa. However, the SOD was in dimer form as revealed by gel filtration chromatography. The purified recombinant enzyme had a pI of 4.5 and exhibited maximal activity at 25°C and pH 7.2. It was stable up to 45°C. The insensitivity of this lactococcal SOD to cyanide and hydrogen peroxide established that it was a MnSOD. Although it has 98% homology to SOD ofL. lactisIL1403, this is the first elucidated structure of lactococcal SOD revealing active sites containing the catalytic manganese coordinated by four ligands (H-27, H-82, D-168, and H-172).

Using Protein Electrophoresis to Investigate the Phylogeny of Velvetleaf (Abutilon theophrasti)

Weed Science ◽  
1988 ◽  
Vol 36 (2) ◽  
pp. 172-175 ◽  
Author(s):  
Steven J. Stegink ◽  
Neal R. Spencer
Keyword(s):  
Superoxide Dismutase ◽  
Enzyme Activity ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Electrophoresis ◽  
Abutilon Theophrasti ◽  
The Other ◽  
Multiple Forms ◽  
Activity Staining ◽  
Native Polyacrylamide Gel Electrophoresis

Native Polyacrylamide gel electrophoresis (PAGE) and enzyme activity staining were used to identify possible progenitor species of velvetleaf (Abutilon theophrastiMedic. # ABUTH). Multiple forms of superoxide dismutase activity were observed in each of the plants surveyed. Three enzyme forms were common to all the species and bio types while one form was different in the velvetleaf biotypes compared to the other species. Multiple forms of peroxidase activity were also detected. The three velvetleaf biotypes possessed identical enzyme forms with minimal similarity to peroxidase forms found in the other species. Multiple forms of esterase activity separated as two nonoverlapping groups. A slowly migrating group was observed in all the velvetleaf biotypes and a more rapidly migrating group characterized the remainingAbutilonspecies. The results of this study indicated that the progenitors of velvetleaf were not among the species surveyed and suggested that the progenitors may no longer be extant.

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