Highly efficient synthesis of beta(1 -> 4)-oligo- and -polysaccharides using a mutant cellulase

S  Fort; V  Boyer; L  Greffe; G  Davies; O  Moroz; L  Christiansen; M  Schulein; S  Cottaz; H  Driguez

doi:10.1021/ja9936520

Highly efficient synthesis of beta(1 -> 4)-oligo- and -polysaccharides using a mutant cellulase

S Fort, V Boyer, L Greffe, G Davies, O Moroz, L Christiansen, M Schulein, S Cottaz, H Driguez

Chemistry

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

Abstract

This report describes an efficient chemoenzymatic synthesis of a variety of regioselectively modified beta(1-->4)-oligo- and -polysaccharides. This successfull approach was based on: (i) the use of a "glycosynthase" which is a Glu-197-Ala nucleophile mutant of the retaining cellulase endoglucanase I (Cel7B) from Humicola insolens and (ii) the rational design of modified acceptor and donor molecules through a careful examination of information given by the X-ray structures of wild type and mutated enzymes. The mutant was able to catalyze, in high yield, the regio- and stereoselective glycosylation of alpha-glycobiosyl fluorides both unsubstituted and modified with various mono- and disaccharide accepters, as well as the polymerization of these donors through a single-step inverting mechanism.

Original language	English
Pages (from-to)	5429-5437
Number of pages	9
Journal	Journal of the American Chemical Society
Volume	122
Issue number	23
DOIs	https://doi.org/10.1021/ja9936520
Publication status	Published - 14 Jun 2000

Keywords

GLYCOSIDASE-CATALYZED SYNTHESIS
N-ACETYLNEURAMINIC ACID
ENZYMATIC-SYNTHESIS
ENDOGLUCANASE-I
ANGSTROM RESOLUTION
MACROMOLECULAR STRUCTURES
FUSARIUM-OXYSPORUM
CRYSTAL-STRUCTURE
GLYCOSYL TRANSFER
OLIGOSACCHARIDES

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10.1021/ja9936520

Cite this

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title = "Highly efficient synthesis of beta(1 -> 4)-oligo- and -polysaccharides using a mutant cellulase",

abstract = "This report describes an efficient chemoenzymatic synthesis of a variety of regioselectively modified beta(1-->4)-oligo- and -polysaccharides. This successfull approach was based on: (i) the use of a {"}glycosynthase{"} which is a Glu-197-Ala nucleophile mutant of the retaining cellulase endoglucanase I (Cel7B) from Humicola insolens and (ii) the rational design of modified acceptor and donor molecules through a careful examination of information given by the X-ray structures of wild type and mutated enzymes. The mutant was able to catalyze, in high yield, the regio- and stereoselective glycosylation of alpha-glycobiosyl fluorides both unsubstituted and modified with various mono- and disaccharide accepters, as well as the polymerization of these donors through a single-step inverting mechanism.",

keywords = "GLYCOSIDASE-CATALYZED SYNTHESIS, N-ACETYLNEURAMINIC ACID, ENZYMATIC-SYNTHESIS, ENDOGLUCANASE-I, ANGSTROM RESOLUTION, MACROMOLECULAR STRUCTURES, FUSARIUM-OXYSPORUM, CRYSTAL-STRUCTURE, GLYCOSYL TRANSFER, OLIGOSACCHARIDES",

author = "S Fort and V Boyer and L Greffe and G Davies and O Moroz and L Christiansen and M Schulein and S Cottaz and H Driguez",

year = "2000",

month = jun,

day = "14",

doi = "10.1021/ja9936520",

language = "English",

volume = "122",

pages = "5429--5437",

journal = "Journal of the American Chemical Society",

issn = "0002-7863",

publisher = "American Chemical Society",

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T1 - Highly efficient synthesis of beta(1 -> 4)-oligo- and -polysaccharides using a mutant cellulase

AU - Fort, S

AU - Boyer, V

AU - Greffe, L

AU - Davies, G

AU - Moroz, O

AU - Christiansen, L

AU - Schulein, M

AU - Cottaz, S

AU - Driguez, H

PY - 2000/6/14

Y1 - 2000/6/14

N2 - This report describes an efficient chemoenzymatic synthesis of a variety of regioselectively modified beta(1-->4)-oligo- and -polysaccharides. This successfull approach was based on: (i) the use of a "glycosynthase" which is a Glu-197-Ala nucleophile mutant of the retaining cellulase endoglucanase I (Cel7B) from Humicola insolens and (ii) the rational design of modified acceptor and donor molecules through a careful examination of information given by the X-ray structures of wild type and mutated enzymes. The mutant was able to catalyze, in high yield, the regio- and stereoselective glycosylation of alpha-glycobiosyl fluorides both unsubstituted and modified with various mono- and disaccharide accepters, as well as the polymerization of these donors through a single-step inverting mechanism.

AB - This report describes an efficient chemoenzymatic synthesis of a variety of regioselectively modified beta(1-->4)-oligo- and -polysaccharides. This successfull approach was based on: (i) the use of a "glycosynthase" which is a Glu-197-Ala nucleophile mutant of the retaining cellulase endoglucanase I (Cel7B) from Humicola insolens and (ii) the rational design of modified acceptor and donor molecules through a careful examination of information given by the X-ray structures of wild type and mutated enzymes. The mutant was able to catalyze, in high yield, the regio- and stereoselective glycosylation of alpha-glycobiosyl fluorides both unsubstituted and modified with various mono- and disaccharide accepters, as well as the polymerization of these donors through a single-step inverting mechanism.

KW - GLYCOSIDASE-CATALYZED SYNTHESIS

KW - N-ACETYLNEURAMINIC ACID

KW - ENZYMATIC-SYNTHESIS

KW - ENDOGLUCANASE-I

KW - ANGSTROM RESOLUTION

KW - MACROMOLECULAR STRUCTURES

KW - FUSARIUM-OXYSPORUM

KW - CRYSTAL-STRUCTURE

KW - GLYCOSYL TRANSFER

KW - OLIGOSACCHARIDES

U2 - 10.1021/ja9936520

DO - 10.1021/ja9936520

M3 - Article

SN - 0002-7863

VL - 122

SP - 5429

EP - 5437

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

IS - 23

ER -