Purification and characterization of a hyperthermostable Mn-superoxide dismutase from Thermus thermophilus HB27

Abstract

Thermostable Mn-dependent catalases are promising enzymes in biotechnological applications. In the present study, a Mn-containing superoxide dismutase of the hyperthermophilic Thermus thermophilus HB27 had been purified and characterized by a two-stage ultrafiltration process after being expressed in E. coli. The enzyme was highly stable at 90°C and retained 57% activity after heat treatment at 100°C for 1 h. The native form of the enzyme was determined as a homotetramer by analytical size exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The final purified enzyme had an isoelectric point of 6.2 and a high α-helical content of 70%, consistent with the theoretical values. This showed that the purified SOD folded with a reasonable secondary structure.

Characterization of the interaction between superoxide dismutase and 2-oxoisovalerate dehydrogenase

“…T. thermophilus is found in hot springs of neutral and alkaline pH, as well as in hot water or in natural waters subjected to thermal pollution and grows optimally at temperatures around 70-75°C (Fuciños et al, 2005). The thermophile produces a number of enzymes of considerable biotechnological interests, such as proteases, phosphatases, catalases, several DNA processing enzymes and lipases, which have been extensively documented (Pantazaki et al, 2002;Fuciños et al, 2005;Song and Zhang, 2008;Fuciños et al, 2011;Liu et al, 2011;Serrano-Posada et al, 2011;Sandoval et al, 2012;Song et al, 2012;Torres et al, 2012;Murugapiran et al, 2013). Although these enzymes play important roles in the thermoadaptation of thermophiles, the pathways mediated by them remain unclear.…”

Unique Characteristics of Recombinant Hybrid Manganese Superoxide Dismutase from Staphylococcus equorum and S. saprophyticus

“…The enzyme has indeed been grouped into narrow and wide pH range SOD. Examples from the first group are SOD from T. thermophilus HB27 (pH 5.0-7.0) [21], Thermothrix sp (pH 8.0-10.0) [8], and Bacillus sp. MHS47 (pH 7.0-8.5) [9].…”

“…Interestingly, few SODs can also be cambialistic, which means that the type of metal installed is exchangeable [20]. Further, SODs demonstrate stability against various environmental stress, particularly those originated from thermophilic or hyperthermophilic microorganisms [8][9][10][11][12][13][14]21]. Thermostable SOD are of interest for application such as sun cream and for its stability during production, formulation, shipping, and storage.…”

A novel membrane based process to isolate recombinant human chemokine receptor CCR3 produced in Escherichia coli

“…Circular dichroism (CD) is a valuable method for the analysis of protein secondary structure, and the far-UV CD spectroscopy from 190 nm to 250 nm can be used to estimate contents of secondary structures [29]. We used the UF purified MBP-CCR3 for CD study, and the CD profile displayed a typical alpha-helix curve with $63% content (Fig.…”