A newly identified glutaminase-free ʟ -asparaginase (ʟ -ASPG86) from the marine bacterium Mesoflavibacter zeaxanthinifaciens
- Title
- A newly identified glutaminase-free ʟ -asparaginase (ʟ -ASPG86) from the marine bacterium Mesoflavibacter zeaxanthinifaciens
- KIOST Author(s)
- Lee, Sujin(이수진); Lee, Young Deuk(이영득); Jo, Eunyoung(조은영); Oh, Chul Hong(오철홍)
- Alternative Author(s)
- 이수진; 이영득; 조은영; Amarin; 오철홍
- Publication Year
- 2017-09-26
- Abstract
- ʟ -Asparaginase (EC 3.5.1.1) is an enzyme involved in asparagine hydrolysis and has the potential to effect leukemic cells and various other cancer cells. We identified the ʟ -asparaginase gene (ʟ -ASPG86) in the genus Mesoflavibacter, which consists of a 1035-bp open reading frame (ORF) encoding 344 amino acids (aa). Following phylogenetic analysis, the deduced amino acid sequence of ʟ -ASPG86 (ʟ -ASPG86) grouped as a type I asparaginase with respective homologs in Escherichia coli and Yersinia pseudotuberculosis. The ʟ -ASPG86 gene was cloned into the pET-16b vector to express the respective protein in E. coli BL21 (DE3) cells. Recombinant ʟ -asparaginase (r-ʟ -ASPG86) showed optimum conditions at 37-40oC, pH 9. Moreover, r-ʟ -ASPG86 did not exhibit glutaminase activity. In the metal ions test, its enzymatic activity was highly improved upon addition of 5 mM manganese (3.97-fold) and magnesium (3.35-fold) compared with the untreated control. The specific activity of r-ʟ -ASPG86 was 687.1 units/mg under optimum conditions (37oC, pH 9 and 5 mM MnSO4).bacter, which consists of a 1035-bp open reading frame (ORF) encoding 344 amino acids (aa). Following phylogenetic analysis, the deduced amino acid sequence of ʟ -ASPG86 (ʟ -ASPG86) grouped as a type I asparaginase with respective homologs in Escherichia coli and Yersinia pseudotuberculosis. The ʟ -ASPG86 gene was cloned into the pET-16b vector to express the respective protein in E. coli BL21 (DE3) cells. Recombinant ʟ -asparaginase (r-ʟ -ASPG86) showed optimum conditions at 37-40oC, pH 9. Moreover, r-ʟ -ASPG86 did not exhibit glutaminase activity. In the metal ions test, its enzymatic activity was highly improved upon addition of 5 mM manganese (3.97-fold) and magnesium (3.35-fold) compared with the untreated control. The specific activity of r-ʟ -ASPG86 was 687.1 units/mg under optimum conditions (37oC, pH 9 and 5 mM MnSO4).
- Bibliographic Citation
- Enzyme Engineering XXIX, pp.97, 2017
- Publisher
- Engineering Conferences International
- Type
- Conference
- Language
- English
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