• Journal of Microbiology and Biotechnology
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• v.24 no.8
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• pp.1096-1104
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• 2014

$\small{L}$-Asparaginase from gram-positive bacteria has been poorly explored. We conducted recombinant overexpression and purification of $\small{L}$-asparaginase from Staphylococcus sp. OJ82 (SoAsn) isolated from Korean fermented seafood to evaluate its biotechnological potential as an antileukemic agent. SoAsn was expressed in Escherichia coli BL21 (DE3) with an estimated molecular mass of 37.5 kDa, determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Consistent with asparaginases in gram-negative bacteria, size-exclusion chromatography determined SoAsn as a homodimer. Interestingly, the optimal temperature of SoAsn was $37^{\circ}C$ and over 90% of activity was retained between $37^{\circ}C$ and $50^{\circ}C$, and its thermal stability range was narrower than that of commercial E. coli $\small{L}$-asparaginase (EcAsn). Both SoAsn and EcAsn were active between pH 9 and 10, although their overall pH-dependent enzyme activities were slightly different. The $K_m$ value of SoAsn was 2.2 mM, which is higher than that of EcAsn. Among eight metals tested for enzyme activity, cobalt and magnesium greatly enhanced the SoAsn and EcAsn activity, respectively. Interestingly, SoAsn retained more than 60% of its activity under 2 M NaCl condition, but the activity of EcAsn was reduced to 48%. Overall, the biochemical characteristics of SoAsn were similar to those of EcAsn, but its kinetics, cofactor requirements, and NaCl tolerance differed from those of EcAsn.

• Journal of Microbiology and Biotechnology
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• v.18 no.11
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• pp.1809-1818
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• 2008

Biotechnological treatment of sulfate- and metal-ions-containing acidic wastewaters from mining and metallurgical activities utilizes sulfate-reducing bacteria to produce sulfide that can subsequently precipitate metal ions. Reducing sulfate at a low pH has several advantages above neutrophilic sulfate reduction. This study describes the effect of sulfide removal on the reactor performance and microbial community in a high-rate sulfidogenic gas-lift bioreactor fed with hydrogen at a controlled internal pH of 5. Under sulfide removal conditions, 99% of the sulfate was converted at a hydraulic retention time of 24 h, reaching a volumetric activity as high as 51 mmol sulfate/l/d. Under nonsulfide removal conditions, <25% of the sulfate was converted at a hydraulic retention time of 24 h reaching volumetric activities of <13 mmol sulfate/l/d. The absence of sulfide removal at a hydraulic retention time of 24 h resulted in an average $H_2S$ concentration of 18.2 mM (584 mg S/I). The incomplete sulfate removal was probably due to sulfide inhibition. Molecular phylogenetic analysis identified 11 separate 16S rRNA bands under sulfide stripping conditions, whereas under nonsulfide removal conditions only 4 separate 16S rRNA bands were found. This shows that a less diverse population was found in the presence of a high sulfide concentration.

• Korean Journal of Microbiology
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• v.53 no.4
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• pp.309-315
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• 2017

Most of acidophiles are found in the various low pH environments and affect to metal cycle through oxidation and reduction reactions. The present study was carried out above 50 strains as acidophiles isolated from acidic soils of exhausted mine and Jeju Gotjawal. Finally, total 19 strains obtained and were tentatively identified based on comparative similarity analysis for 16S rRNA gene sequence and physiological characterizations. These isolates belonged to Gammaproteobacteria (6 strains), Actinobacteria (5 strains), Betaproteobacteria (4 strains), Alphaproteobacteria (2 strains), and Bacilli (2 strains). We observed that these isolates can grow under low pH culture condition. This case study for analysis physiological characterizations of indigenous microorganisms in acidic soil might provide basic information on useful application.