Identification of amino acid residues that determine the substrate preference of 1,3-β-galactosyl-N-acetylhexosamine phosphorylase

Mamoru Nishimoto; Masafumi Hidaka; Masahiro Nakajima; Shinya Fushinobu; Motomitsu Kitaoka

doi:10.1016/j.molcatb.2011.09.004

Identification of amino acid residues that determine the substrate preference of 1,3-β-galactosyl-N-acetylhexosamine phosphorylase

Mamoru Nishimoto, Masafumi Hidaka, Masahiro Nakajima, Shinya Fushinobu, Motomitsu Kitaoka

AGR - Agricultural Chemistry

Research output: Contribution to journal › Article › peer-review

6 Citations (Scopus)

Abstract

Three amino acid residues of 1,3-β-galactosyl-N-acetylhexosamine phosphorylase (GalHexNAcP) were assigned as the determinants of substrate preference for galacto-N-biose (GNB) and lacto-N-biose I (LNB) based on the three-dimensional structure of the protein. Mutants of GalHexNAcP from Bifidobacterium longum, which acts similarly on both GNB and LNB, were constructed and characterized. V162T mutation led to an increase in the selectivity on GNB. P161S and S336A mutations independently enhanced the selectivity on LNB. The alignment of amino acid sequences suggests that the activities of most homologous sequences are predictable by comparing the corresponding three residues. .

Original language	English
Pages (from-to)	97-102
Number of pages	6
Journal	Journal of Molecular Catalysis B: Enzymatic
Volume	74
Issue number	1-2
DOIs	https://doi.org/10.1016/j.molcatb.2011.09.004
Publication status	Published - 2012 Jan

Keywords

1,3-β-Galactosyl-N-acetylhexosamine phosphorylase
Determinant residue of substrate preference
Galacto-N-biose
Glycoside hydrolase family 112
Lacto-N-biose I

ASJC Scopus subject areas

Catalysis
Bioengineering
Biochemistry
Process Chemistry and Technology

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10.1016/j.molcatb.2011.09.004

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title = "Identification of amino acid residues that determine the substrate preference of 1,3-β-galactosyl-N-acetylhexosamine phosphorylase",

abstract = "Three amino acid residues of 1,3-β-galactosyl-N-acetylhexosamine phosphorylase (GalHexNAcP) were assigned as the determinants of substrate preference for galacto-N-biose (GNB) and lacto-N-biose I (LNB) based on the three-dimensional structure of the protein. Mutants of GalHexNAcP from Bifidobacterium longum, which acts similarly on both GNB and LNB, were constructed and characterized. V162T mutation led to an increase in the selectivity on GNB. P161S and S336A mutations independently enhanced the selectivity on LNB. The alignment of amino acid sequences suggests that the activities of most homologous sequences are predictable by comparing the corresponding three residues. .",

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author = "Mamoru Nishimoto and Masafumi Hidaka and Masahiro Nakajima and Shinya Fushinobu and Motomitsu Kitaoka",

year = "2012",

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T1 - Identification of amino acid residues that determine the substrate preference of 1,3-β-galactosyl-N-acetylhexosamine phosphorylase

AU - Nishimoto, Mamoru

AU - Hidaka, Masafumi

AU - Nakajima, Masahiro

AU - Fushinobu, Shinya

AU - Kitaoka, Motomitsu

PY - 2012/1

Y1 - 2012/1

N2 - Three amino acid residues of 1,3-β-galactosyl-N-acetylhexosamine phosphorylase (GalHexNAcP) were assigned as the determinants of substrate preference for galacto-N-biose (GNB) and lacto-N-biose I (LNB) based on the three-dimensional structure of the protein. Mutants of GalHexNAcP from Bifidobacterium longum, which acts similarly on both GNB and LNB, were constructed and characterized. V162T mutation led to an increase in the selectivity on GNB. P161S and S336A mutations independently enhanced the selectivity on LNB. The alignment of amino acid sequences suggests that the activities of most homologous sequences are predictable by comparing the corresponding three residues. .

AB - Three amino acid residues of 1,3-β-galactosyl-N-acetylhexosamine phosphorylase (GalHexNAcP) were assigned as the determinants of substrate preference for galacto-N-biose (GNB) and lacto-N-biose I (LNB) based on the three-dimensional structure of the protein. Mutants of GalHexNAcP from Bifidobacterium longum, which acts similarly on both GNB and LNB, were constructed and characterized. V162T mutation led to an increase in the selectivity on GNB. P161S and S336A mutations independently enhanced the selectivity on LNB. The alignment of amino acid sequences suggests that the activities of most homologous sequences are predictable by comparing the corresponding three residues. .

KW - 1,3-β-Galactosyl-N-acetylhexosamine phosphorylase

KW - Determinant residue of substrate preference

KW - Galacto-N-biose

KW - Glycoside hydrolase family 112

KW - Lacto-N-biose I

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Identification of amino acid residues that determine the substrate preference of 1,3-β-galactosyl-N-acetylhexosamine phosphorylase

Abstract

Keywords

ASJC Scopus subject areas

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