Biochemical characterization of L-Asparaginase in NaCl-tolerant Staphylococcus sp. OJ82 isolated from fermented seafood

Sangwon Han, Jaejoon Jung, Woojun Park

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

L-Asparaginase from gram-positive bacteria has been poorly explored. We conducted recombinant overexpression and purification of 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°C and over 90% of activity was retained between 37°C and 50°C, and its thermal stability range was narrower than that of commercial E. coli 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 Km 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.

Original languageEnglish
Pages (from-to)1096-1104
Number of pages9
JournalJournal of Microbiology and Biotechnology
Volume24
Issue number8
DOIs
Publication statusPublished - 2014 May 27

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Asparaginase
Seafood
Staphylococcus
Escherichia coli
Gram-Positive Bacteria
Enzymes
Cobalt
Gram-Negative Bacteria
Sodium Dodecyl Sulfate
Magnesium
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Hot Temperature
Metals

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Biochemical characterization of L-Asparaginase in NaCl-tolerant Staphylococcus sp. OJ82 isolated from fermented seafood. / Han, Sangwon; Jung, Jaejoon; Park, Woojun.

In: Journal of Microbiology and Biotechnology, Vol. 24, No. 8, 27.05.2014, p. 1096-1104.

Research output: Contribution to journalArticle

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abstract = "L-Asparaginase from gram-positive bacteria has been poorly explored. We conducted recombinant overexpression and purification of 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°C and over 90{\%} of activity was retained between 37°C and 50°C, and its thermal stability range was narrower than that of commercial E. coli 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 Km 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.",
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