Hydrolysis of α- and β-d-glucosyl fluoride by individual glucosidases

new evidence for separately controlled "plastic" and "conserved" phases in glycosylase catalysis

Hirokazu Matsui, Yoshimasa Tanaka, Curtis F. Brewer, John S. Blanchard, Edward J. Hehre

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

α-Glucosidases from sugar beet seed and ungerminated rice catalyzed the hydrolysis of β-d-glucopyranosyl fluoride to form α-d-glucose. The reactions were slow, with V/K = 11-15 × 10-3 or ∼ 1-2% of that for hydrolysis of p-nitrophenyl α-d-glucopyranoside, but were not due to any impurity in the substrate or to contaminating β-glucosidase or glucoamylase. Furthermore, almond β-glucosidase promoted hydrolysis of α-d-glucosyl fluoride to form β-d-glucose at an exceedingly low rate, V/K = 4 × 10-4. This weak reaction did not stem from any impurity in the substrate or to contamination with α-glucosidase or glucoamylase, but it was partly (∼ 20%) attributable to a trace of accompanying trehalase. That all three glucosidases acted upon both α- and β-d-glucosyl fluoride, albeit at low efficiency with the disfavored anomer, reflects the previously demonstrated ability of each enzyme's catalytic groups to respond flexibly to substrates of different types. That the disfavored d-glucosyl fluoride in each case was converted into a product of the same configuration as from enitols or favored d-glucosyl substrates provides additional evidence for the two-step nature of the chemical mechanisms of glucosidases, in which the stereochemistry of water attack on the enzyme-stabilized oxocarbonium ion is strictly maintained, regardless of the initial anomeric configuration of the substrate.

Original languageEnglish (US)
Pages (from-to)45-56
Number of pages12
JournalCarbohydrate Research
Volume250
Issue number1
DOIs
StatePublished - Dec 16 1993

Fingerprint

Glucosidases
Catalysis
Plastics
Hydrolysis
Glucan 1,4-alpha-Glucosidase
Substrates
Trehalase
Impurities
Sugar beets
Glucose
Stereochemistry
Beta vulgaris
Enzymes
Fluorides
Seed
glucosyl fluoride
Seeds
Contamination
Ions
Water

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Organic Chemistry

Cite this

@article{78b8105920384f74ada82856f30cbdc5,
title = "Hydrolysis of α- and β-d-glucosyl fluoride by individual glucosidases: new evidence for separately controlled {"}plastic{"} and {"}conserved{"} phases in glycosylase catalysis",
abstract = "α-Glucosidases from sugar beet seed and ungerminated rice catalyzed the hydrolysis of β-d-glucopyranosyl fluoride to form α-d-glucose. The reactions were slow, with V/K = 11-15 × 10-3 or ∼ 1-2{\%} of that for hydrolysis of p-nitrophenyl α-d-glucopyranoside, but were not due to any impurity in the substrate or to contaminating β-glucosidase or glucoamylase. Furthermore, almond β-glucosidase promoted hydrolysis of α-d-glucosyl fluoride to form β-d-glucose at an exceedingly low rate, V/K = 4 × 10-4. This weak reaction did not stem from any impurity in the substrate or to contamination with α-glucosidase or glucoamylase, but it was partly (∼ 20{\%}) attributable to a trace of accompanying trehalase. That all three glucosidases acted upon both α- and β-d-glucosyl fluoride, albeit at low efficiency with the disfavored anomer, reflects the previously demonstrated ability of each enzyme's catalytic groups to respond flexibly to substrates of different types. That the disfavored d-glucosyl fluoride in each case was converted into a product of the same configuration as from enitols or favored d-glucosyl substrates provides additional evidence for the two-step nature of the chemical mechanisms of glucosidases, in which the stereochemistry of water attack on the enzyme-stabilized oxocarbonium ion is strictly maintained, regardless of the initial anomeric configuration of the substrate.",
author = "Hirokazu Matsui and Yoshimasa Tanaka and Brewer, {Curtis F.} and Blanchard, {John S.} and Hehre, {Edward J.}",
year = "1993",
month = "12",
day = "16",
doi = "10.1016/0008-6215(93)84153-W",
language = "English (US)",
volume = "250",
pages = "45--56",
journal = "Carbohydrate Research",
issn = "0008-6215",
publisher = "Elsevier BV",
number = "1",

}

TY - JOUR

T1 - Hydrolysis of α- and β-d-glucosyl fluoride by individual glucosidases

T2 - new evidence for separately controlled "plastic" and "conserved" phases in glycosylase catalysis

AU - Matsui, Hirokazu

AU - Tanaka, Yoshimasa

AU - Brewer, Curtis F.

AU - Blanchard, John S.

AU - Hehre, Edward J.

PY - 1993/12/16

Y1 - 1993/12/16

N2 - α-Glucosidases from sugar beet seed and ungerminated rice catalyzed the hydrolysis of β-d-glucopyranosyl fluoride to form α-d-glucose. The reactions were slow, with V/K = 11-15 × 10-3 or ∼ 1-2% of that for hydrolysis of p-nitrophenyl α-d-glucopyranoside, but were not due to any impurity in the substrate or to contaminating β-glucosidase or glucoamylase. Furthermore, almond β-glucosidase promoted hydrolysis of α-d-glucosyl fluoride to form β-d-glucose at an exceedingly low rate, V/K = 4 × 10-4. This weak reaction did not stem from any impurity in the substrate or to contamination with α-glucosidase or glucoamylase, but it was partly (∼ 20%) attributable to a trace of accompanying trehalase. That all three glucosidases acted upon both α- and β-d-glucosyl fluoride, albeit at low efficiency with the disfavored anomer, reflects the previously demonstrated ability of each enzyme's catalytic groups to respond flexibly to substrates of different types. That the disfavored d-glucosyl fluoride in each case was converted into a product of the same configuration as from enitols or favored d-glucosyl substrates provides additional evidence for the two-step nature of the chemical mechanisms of glucosidases, in which the stereochemistry of water attack on the enzyme-stabilized oxocarbonium ion is strictly maintained, regardless of the initial anomeric configuration of the substrate.

AB - α-Glucosidases from sugar beet seed and ungerminated rice catalyzed the hydrolysis of β-d-glucopyranosyl fluoride to form α-d-glucose. The reactions were slow, with V/K = 11-15 × 10-3 or ∼ 1-2% of that for hydrolysis of p-nitrophenyl α-d-glucopyranoside, but were not due to any impurity in the substrate or to contaminating β-glucosidase or glucoamylase. Furthermore, almond β-glucosidase promoted hydrolysis of α-d-glucosyl fluoride to form β-d-glucose at an exceedingly low rate, V/K = 4 × 10-4. This weak reaction did not stem from any impurity in the substrate or to contamination with α-glucosidase or glucoamylase, but it was partly (∼ 20%) attributable to a trace of accompanying trehalase. That all three glucosidases acted upon both α- and β-d-glucosyl fluoride, albeit at low efficiency with the disfavored anomer, reflects the previously demonstrated ability of each enzyme's catalytic groups to respond flexibly to substrates of different types. That the disfavored d-glucosyl fluoride in each case was converted into a product of the same configuration as from enitols or favored d-glucosyl substrates provides additional evidence for the two-step nature of the chemical mechanisms of glucosidases, in which the stereochemistry of water attack on the enzyme-stabilized oxocarbonium ion is strictly maintained, regardless of the initial anomeric configuration of the substrate.

UR - http://www.scopus.com/inward/record.url?scp=0027759648&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027759648&partnerID=8YFLogxK

U2 - 10.1016/0008-6215(93)84153-W

DO - 10.1016/0008-6215(93)84153-W

M3 - Article

VL - 250

SP - 45

EP - 56

JO - Carbohydrate Research

JF - Carbohydrate Research

SN - 0008-6215

IS - 1

ER -