Preparation and use of α-maltosyl fluoride as a substrate by beta amylase

Dorothy S. Genghof, Curtis F. Brewer, Edward J. Hehre

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The preparation of pure (amorphous) α-maltosyl fluoride is described. A modification of the procedure of Brauns was used to obtain analytically pure, crystalline hepta-O-acetyl-α-maltosyl fluoride, the structure of which was assigned by 19F- and 1H-n.m.r. spectroscopy. α-Maltosyl fluoride was obtained by deacetylating the heptaacetate. It behaved as a single compound on thin-layer and paper chromatography, and was essentially completely hydrolyzed to maltose and hydrogen fluoride by 0.01M sulfuric acid in 10 min at 100°. Crystalline beta amylase, likewise, catalyzed essentially complete hydrolysis of α-maltosyl fluoride to give maltose and hydrogen fluoride. The rates of hydrolysis catalyzed by beta amylase preparations from sweet potatoes and soybeans acting on a range of concentrations of the substrate produced linear curves for the relationship, 1/v vs 1/S; reaction constants for crystalline, sweet-potato enzyme were Km 3.6 mM and Vmax ∼2 μmol/min/mg. The finding that α-maltosyl fluoride is hydrolyzed 30-60 times faster than maltotriose demonstrates for the first time that beta amylase is capable of effecting hydrolysis at an appreciable rate of a substrate having only two D-glucose residues.

Original languageEnglish (US)
Pages (from-to)291-299
Number of pages9
JournalCarbohydrate Research
Volume60
Issue number2
StatePublished - Mar 1978

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beta-Amylase
Fluorides
Hydrofluoric Acid
Ipomoea batatas
Substrates
Hydrolysis
Maltose
Crystalline materials
Paper Chromatography
Thin Layer Chromatography
Chromatography
Soybeans
Rate constants
Spectrum Analysis
Spectroscopy
Glucose
Enzymes

ASJC Scopus subject areas

  • Organic Chemistry
  • Molecular Biology
  • Biochemistry

Cite this

Preparation and use of α-maltosyl fluoride as a substrate by beta amylase. / Genghof, Dorothy S.; Brewer, Curtis F.; Hehre, Edward J.

In: Carbohydrate Research, Vol. 60, No. 2, 03.1978, p. 291-299.

Research output: Contribution to journalArticle

Genghof, Dorothy S. ; Brewer, Curtis F. ; Hehre, Edward J. / Preparation and use of α-maltosyl fluoride as a substrate by beta amylase. In: Carbohydrate Research. 1978 ; Vol. 60, No. 2. pp. 291-299.
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