Identification of lacto-N-Biose I phosphorylase from Vibrio vulnificus CMCP6.

Applied and Environmental Microbiology
Masahiro Nakajima, Motomitsu Kitaoka

Abstract

A beta-1,3-galactosyl-N-acetylhexosamine phosphorylase (GalGlyNAcP) homolog gene was cloned from Vibrio vulnificus CMCP6. In synthetic reactions, the recombinant enzyme acted only with GlcNAc and GalNAc as acceptors in the presence of alpha-d-galactose-1-phosphate as a donor to form lacto-N-biose I (LNB) (Galbeta1 --> 3GlcNAc) and galacto-N-biose (GNB) (Galbeta1 --> 3GalNAc), respectively. GlcNAc was a much better acceptor than GalNAc. The enzyme also phosphorolysed LNB faster than it phosphorolysed GNB, and the k(cat)/K(m) for LNB was approximately 60 times higher than the k(cat)/K(m) for GNB. This result indicated that the enzyme was remarkably different from GalGlyNAcP from Bifidobacterium longum, which has similar activities with LNB and GNB, and GalGlyNAcP from Clostridium perfringens, which is a GNB-specific enzyme. The enzyme is the first LNB-specific enzyme that has been found and was designated lacto-N-biose I phosphorylase. The discovery of an LNB-specific GalGlyNAcP resulted in recategorization of bifidobacterial GalGlyNAcPs as galacto-N-biose/lacto-N-biose I phosphorylases.

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Citations

Jan 10, 2009·Applied Microbiology and Biotechnology·Masahiro NakajimaMotomitsu Kitaoka
Aug 3, 2011·Applied Microbiology and Biotechnology·Takanori NihiraMotomitsu Kitaoka
Jan 7, 2009·The Journal of Biological Chemistry·Masafumi HidakaShinya Fushinobu
Sep 26, 2013·Protein Engineering, Design & Selection : PEDS·Yoshiyuki KoyamaMotomitsu Kitaoka
Dec 15, 2010·Bioscience, Biotechnology, and Biochemistry·Shinya Fushinobu
May 8, 2009·Bioscience, Biotechnology, and Biochemistry·Masashi KiyoharaKenji Yamamoto
Mar 22, 2014·PloS One·Masahiro NakajimaMotomitsu Kitaoka
Aug 12, 2010·Bioscience, Biotechnology, and Biochemistry·Masahiro NakajimaMotomitsu Kitaoka
Nov 25, 2011·Enzyme and Microbial Technology·Chao ChenDesmet Tom
Aug 22, 2015·Applied Microbiology and Biotechnology·Motomitsu Kitaoka
Feb 25, 2018·Applied Microbiology and Biotechnology·Rachel Chen
Sep 18, 2013·The Journal of Biological Chemistry·Simon LadevèzeGabrielle Potocki-Véronèse

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