Molecular basis of the amylose-like polymer formation catalyzed by Neisseria polysaccharea amylosucrase

Cécile Albenne; Lars Skov; Osman Asghar Mirza; Michael Gajhede; Georges Feller; Salvino D'Amico; Gwénaëlle André; Gabrielle Potocki-Véronèse; Bart A van der Veen; Pierre Monsan; Magali Remaud-Simeon

doi:10.1074/jbc.M309891200

Molecular basis of the amylose-like polymer formation catalyzed by Neisseria polysaccharea amylosucrase

Cécile Albenne, Lars Skov, Osman Asghar Mirza, Michael Gajhede, Georges Feller, Salvino D'Amico, Gwénaëlle André, Gabrielle Potocki-Véronèse, Bart A van der Veen, Pierre Monsan, Magali Remaud-Simeon

86 Citations (Scopus)

Abstract

Amylosucrase from Neisseria polysaccharea is a remarkable transglucosidase from family 13 of the glycoside-hydrolases that synthesizes an insoluble amylose-like polymer from sucrose in the absence of any primer. Amylosucrase shares strong structural similarities with alpha-amylases. Exactly how this enzyme catalyzes the formation of alpha-1,4-glucan and which structural features are involved in this unique functionality existing in family 13 are important questions still not fully answered. Here, we provide evidence that amylosucrase initializes polymer formation by releasing, through sucrose hydrolysis, a glucose molecule that is subsequently used as the first acceptor molecule. Maltooligosaccharides of increasing size were produced and successively elongated at their nonreducing ends until they reached a critical size and concentration, causing precipitation. The ability of amylosucrase to bind and to elongate maltooligosaccharides is notably due to the presence of key residues at the OB1 acceptor binding site that contribute strongly to the guidance (Arg415, subsite +4) and the correct positioning (Asp394 and Arg446, subsite +1) of acceptor molecules. On the other hand, Arg226 (subsites +2/+3) limits the binding of maltooligosaccharides, resulting in the accumulation of small products (G to G3) in the medium. A remarkable mutant (R226A), activated by the products it forms, was generated. It yields twice as much insoluble glucan as the wild-type enzyme and leads to the production of lower quantities of by-products.

Original language	English
Journal	Journal of Biological Chemistry
Volume	279
Issue number	1
Pages (from-to)	726-34
Number of pages	9
ISSN	0021-9258
DOIs	https://doi.org/10.1074/jbc.M309891200
Publication status	Published - 2 Jan 2004

Keywords

Amino Acid Sequence
Amino Acid Substitution
Binding Sites
Conserved Sequence
Glucosyltransferases
Models, Molecular
Mutagenesis, Site-Directed
Neisseria
Oligosaccharides
Protein Conformation
Recombinant Proteins
Sequence Alignment
Sequence Homology, Amino Acid
Sucrose

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10.1074/jbc.M309891200

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title = "Molecular basis of the amylose-like polymer formation catalyzed by Neisseria polysaccharea amylosucrase",

abstract = "Amylosucrase from Neisseria polysaccharea is a remarkable transglucosidase from family 13 of the glycoside-hydrolases that synthesizes an insoluble amylose-like polymer from sucrose in the absence of any primer. Amylosucrase shares strong structural similarities with alpha-amylases. Exactly how this enzyme catalyzes the formation of alpha-1,4-glucan and which structural features are involved in this unique functionality existing in family 13 are important questions still not fully answered. Here, we provide evidence that amylosucrase initializes polymer formation by releasing, through sucrose hydrolysis, a glucose molecule that is subsequently used as the first acceptor molecule. Maltooligosaccharides of increasing size were produced and successively elongated at their nonreducing ends until they reached a critical size and concentration, causing precipitation. The ability of amylosucrase to bind and to elongate maltooligosaccharides is notably due to the presence of key residues at the OB1 acceptor binding site that contribute strongly to the guidance (Arg415, subsite +4) and the correct positioning (Asp394 and Arg446, subsite +1) of acceptor molecules. On the other hand, Arg226 (subsites +2/+3) limits the binding of maltooligosaccharides, resulting in the accumulation of small products (G to G3) in the medium. A remarkable mutant (R226A), activated by the products it forms, was generated. It yields twice as much insoluble glucan as the wild-type enzyme and leads to the production of lower quantities of by-products.",

keywords = "Amino Acid Sequence, Amino Acid Substitution, Binding Sites, Conserved Sequence, Glucosyltransferases, Models, Molecular, Mutagenesis, Site-Directed, Neisseria, Oligosaccharides, Protein Conformation, Recombinant Proteins, Sequence Alignment, Sequence Homology, Amino Acid, Sucrose",

author = "C{\'e}cile Albenne and Lars Skov and Mirza, {Osman Asghar} and Michael Gajhede and Georges Feller and Salvino D'Amico and Gw{\'e}na{\"e}lle Andr{\'e} and Gabrielle Potocki-V{\'e}ron{\`e}se and {van der Veen}, {Bart A} and Pierre Monsan and Magali Remaud-Simeon",

year = "2004",

month = jan,

day = "2",

doi = "10.1074/jbc.M309891200",

language = "English",

volume = "279",

pages = "726--34",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

publisher = "American Society for Biochemistry and Molecular Biology, Inc.",

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TY - JOUR

T1 - Molecular basis of the amylose-like polymer formation catalyzed by Neisseria polysaccharea amylosucrase

AU - Albenne, Cécile

AU - Skov, Lars

AU - Mirza, Osman Asghar

AU - Gajhede, Michael

AU - Feller, Georges

AU - D'Amico, Salvino

AU - André, Gwénaëlle

AU - Potocki-Véronèse, Gabrielle

AU - van der Veen, Bart A

AU - Monsan, Pierre

AU - Remaud-Simeon, Magali

PY - 2004/1/2

Y1 - 2004/1/2

N2 - Amylosucrase from Neisseria polysaccharea is a remarkable transglucosidase from family 13 of the glycoside-hydrolases that synthesizes an insoluble amylose-like polymer from sucrose in the absence of any primer. Amylosucrase shares strong structural similarities with alpha-amylases. Exactly how this enzyme catalyzes the formation of alpha-1,4-glucan and which structural features are involved in this unique functionality existing in family 13 are important questions still not fully answered. Here, we provide evidence that amylosucrase initializes polymer formation by releasing, through sucrose hydrolysis, a glucose molecule that is subsequently used as the first acceptor molecule. Maltooligosaccharides of increasing size were produced and successively elongated at their nonreducing ends until they reached a critical size and concentration, causing precipitation. The ability of amylosucrase to bind and to elongate maltooligosaccharides is notably due to the presence of key residues at the OB1 acceptor binding site that contribute strongly to the guidance (Arg415, subsite +4) and the correct positioning (Asp394 and Arg446, subsite +1) of acceptor molecules. On the other hand, Arg226 (subsites +2/+3) limits the binding of maltooligosaccharides, resulting in the accumulation of small products (G to G3) in the medium. A remarkable mutant (R226A), activated by the products it forms, was generated. It yields twice as much insoluble glucan as the wild-type enzyme and leads to the production of lower quantities of by-products.

AB - Amylosucrase from Neisseria polysaccharea is a remarkable transglucosidase from family 13 of the glycoside-hydrolases that synthesizes an insoluble amylose-like polymer from sucrose in the absence of any primer. Amylosucrase shares strong structural similarities with alpha-amylases. Exactly how this enzyme catalyzes the formation of alpha-1,4-glucan and which structural features are involved in this unique functionality existing in family 13 are important questions still not fully answered. Here, we provide evidence that amylosucrase initializes polymer formation by releasing, through sucrose hydrolysis, a glucose molecule that is subsequently used as the first acceptor molecule. Maltooligosaccharides of increasing size were produced and successively elongated at their nonreducing ends until they reached a critical size and concentration, causing precipitation. The ability of amylosucrase to bind and to elongate maltooligosaccharides is notably due to the presence of key residues at the OB1 acceptor binding site that contribute strongly to the guidance (Arg415, subsite +4) and the correct positioning (Asp394 and Arg446, subsite +1) of acceptor molecules. On the other hand, Arg226 (subsites +2/+3) limits the binding of maltooligosaccharides, resulting in the accumulation of small products (G to G3) in the medium. A remarkable mutant (R226A), activated by the products it forms, was generated. It yields twice as much insoluble glucan as the wild-type enzyme and leads to the production of lower quantities of by-products.

KW - Amino Acid Sequence

KW - Amino Acid Substitution

KW - Binding Sites

KW - Conserved Sequence

KW - Glucosyltransferases

KW - Models, Molecular

KW - Mutagenesis, Site-Directed

KW - Neisseria

KW - Oligosaccharides

KW - Protein Conformation

KW - Recombinant Proteins

KW - Sequence Alignment

KW - Sequence Homology, Amino Acid

KW - Sucrose

U2 - 10.1074/jbc.M309891200

DO - 10.1074/jbc.M309891200

M3 - Journal article

C2 - 14570882

SN - 0021-9258

VL - 279

SP - 726

EP - 734

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 1

ER -

Molecular basis of the amylose-like polymer formation catalyzed by Neisseria polysaccharea amylosucrase

Abstract

Keywords

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