Abstract
A putative recombinant β-galactosidase from Deinococcus geothermalis was purified as a single 79 kDa band of 42 U activity/mg using His-Trap affinity chromatography. The molecular mass of the native enzyme was a 158 kDa dimer. The catalytic residues E151 and E325 of β-galactosidase from D. geothermalis were conserved in all aligned GH family 42 β-galactosidases, indicating that this enzyme is also a GH family 42 β-galactosidase. Maximal activity of the enzyme was at pH 6.5 and 60°C. It has a unique hydrolytic activity for p-nitrophenyl(pNP)-β-d-galactopyranoside (k cat/K m = 69 s−1 mM−1), pNP-β-d-fucopyranoside (13), oNP-β-d-galactopyranoside (9.5), oNP-β-d-fucopyranoside (2.6), lactose (0.97), and pNP-α-l-arabinopyranoside (0.78), whereas no activity, or less than 2% of the pNP-β-d-galactopyranoside activity, for other pNP- and oNP- glycosides.
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This study was carried out with the support of the 21C Frontier Project for Microbial Genomics, Ministry of Education, Science and Technology.
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Lee, JH., Kim, YS., Yeom, SJ. et al. Characterization of a glycoside hydrolase family 42 β-galactosidase from Deinococcus geothermalis . Biotechnol Lett 33, 577–583 (2011). https://doi.org/10.1007/s10529-010-0459-6
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DOI: https://doi.org/10.1007/s10529-010-0459-6