Expression and Purification of Recombinant Superoxide Dismutase (PaSOD) from Psychromonas arctica in Escherichia coli

Na, Ju-Mee; Im, Hana; Lee, Kyung‐Hee

doi:10.5012/bkcs.2011.32.7.2405

Cited by 6 publications

(6 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The low pH optima of both Ye SODs may be of significance under acidic conditions such as those encountered inside the macrophages and eventually help in the dissemination of Y. enterocolitica in the host by sequestering the organism in these cells. At low temperatures, reduction in the cellular metabolic rate causes accumulation of electrons in the respiratory chain that leads to overproduction of reactive oxygen species [56]. Y. enterocolitica being a psychrotrophic food-borne pathogen, the low temperature optima of Ye SODs may help in neutralizing the excess oxygen radicals produced during its growth at lower temperatures in refrigerated foods [57].…”

Section: Discussionmentioning

confidence: 99%

Detection, Distribution and Characterization of Novel Superoxide Dismutases from Yersinia enterocolitica Biovar 1A

2013

View full text Add to dashboard Cite

BackgroundSuperoxide dismutases (SODs) cause dismutation of superoxide radicals to hydrogen peroxide and oxygen. Besides protecting the cells against oxidative damage by endogenously generated oxygen radicals, SODs play an important role in intraphagocytic survival of pathogenic bacteria. The complete genome sequences of Yersinia enterocolitica strains show presence of three different sod genes. However, not much is known about the types of SODs present in Y. enterocolitica, their characteristics and role in virulence and intraphagocytic survival of this organism.Methodology/Principal FindingsThis study reports detection and distribution of the three superoxide dismutase (sodA, sodB and sodC) genes in 59 strains of Y. enterocolitica and related species. The majority (94%) of the strains carried all three genes and constitutive expression of sodA and sodB was detected in 88% of the strains. Expression of sodC was not observed in any of the strains. The sodA, sodB and sodC genes of Y. enterocolitica were cloned in pET28a (+) vector. Recombinant SodA (82 kDa) and SodB (21 kDa) were expressed as homotetramer and monomer respectively, and showed activity over a broad range of pH (3.0–8.0) and temperature (4–70°C). SodA and SodB showed optimal activity at 4°C under acidic pH of 6.0 and 4.0 respectively. The secondary structures of recombinant SodA and SodB were studied using circular dichroism. Production of YeSodC was not observed even after cloning and expression in E. coli BL21(DE3) cells. A SodA− SodB− Escherichia coli strain which was unable to grow in medium supplemented with paraquat showed normal growth after complementation with Y. enterocolitica SodA or SodB.Conclusions/SignificanceThis is the first report on the distribution and characterization of superoxide dismutases from Y. enterocolitica. The low pH optima of both SodA and SodB encoded by Y. enterocolitica seem to implicate their role in acidic environments such as the intraphagocytic vesicles.

show abstract

Section: Discussionmentioning

confidence: 99%

Detection, Distribution and Characterization of Novel Superoxide Dismutases from Yersinia enterocolitica Biovar 1A

2013

View full text Add to dashboard Cite

show abstract

“…SODs exist in a wide variety of organisms, including fish, goat, human, boar, and cold‐active SODs from Pseudoalteromonas haloplanktis , Marinomonas sp. NJ522, and Psychromonas arctica . SODs play an important role in the protection of the cells against the harmful environment of reactive oxygen species (ROS) and other superoxide free radicals, such as the superoxide radical anion (O 2 − ), nitric oxide (NO), and hydroxyl radical (OH) .…”

Section: Introductionmentioning

confidence: 99%

Cloning, expression and biochemical characterization of recombinant superoxide dismutase from Antarctic psychrophilic bacterium Pseudoalteromonas sp. ANT506

Wang

Hou

et al. 2015

J. Basic Microbiol.

View full text Add to dashboard Cite

In this study, a superoxide dismutase gene (PsSOD) from Pseudoalteromonas sp. ANT506 was cloned and over expressed in Escherichia coli. The PsSOD has an open reading frame of 582 bp with a putative product of 193 amino acid residue and an estimated molecular size of 21.4 kDa. His-tagged PsSOD was subsequently purified 12.6-fold by Ni-affinity chromatography and the yield of 22.9%. The characterization of the purified rPsSOD exhibited maximum activity at 30 °C and pH 8.0. The enzyme exhibited 13.9% activity at 0 °C and had high-thermo lability at higher than 50 °C. rPsSOD exhibited well capability to 2.5 M NaCl (62.4%). These results indicated that rPsSOD exhibited special catalytic properties.

show abstract

“…Identification of genes encoding CA SOD synthesis is very seldom published. Until now, such identification has been carried out in bacterial strains Aliivibrio salmonicida (Fe-SOD, sodB) and Psychromonas arctica (Pedersen et al 2009;Na et al 2011).…”

Section: Discussionmentioning

confidence: 99%

Production, purification, and characterization of a novel cold-active superoxide dismutase from the Antarctic strain Aspergillus glaucus 363

et al. 2016

View full text Add to dashboard Cite

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 × 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.

show abstract

Expression and Purification of Recombinant Superoxide Dismutase (PaSOD) from Psychromonas arctica in Escherichia coli

Cited by 6 publications

References 26 publications

Detection, Distribution and Characterization of Novel Superoxide Dismutases from Yersinia enterocolitica Biovar 1A

Detection, Distribution and Characterization of Novel Superoxide Dismutases from Yersinia enterocolitica Biovar 1A

Cloning, expression and biochemical characterization of recombinant superoxide dismutase from Antarctic psychrophilic bacterium Pseudoalteromonas sp. ANT506

Production, purification, and characterization of a novel cold-active superoxide dismutase from the Antarctic strain Aspergillus glaucus 363

Contact Info

Product

Resources

About