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Partial purification and characterization of alpha-glucosidase from Pseudomonas fluorescens W

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Abstract

The α-glucosidase (α-d-glucoside glucohydrolase, EC 3.2.1.20) of Pseudomonas fluorescens W was partially purified by (NH4)2SO4 fractionation, Sephadex G-200 and DEAE-cellulose column chromatography. The enzyme showed great specificity for maltose hydrolysis, with very little action against polymeric forms. Sucrose, isomaltose, α-methylglucoside, and maltobionic acid were not hydrolyzed. Turanose was a strong competitive inhibitor, and glucose a weaker one. Tris (2-amino-2-hydroxymethylpropan-1:3-diol) inhibited enzyme activity significantly only at alkaline pH. Mercuric, cupric, and silver cations strongly inhibited, and EDTA (ethylenediaminetetraacetate) weakly inhibited the enzyme. The isolated enzyme was rather unstable even at 4° C, and was destroyed by freezing and lyophilization. Inositol and albumin had a slightly protective effect. Sulfhydryl-binding reagents strongly inhibited the enzyme.

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Abbreviations

PNPG:

paranitrophenyl-α-d-glucoside

PCMB:

parachloromercuribenzoate

DEAE:

diethylaminoethyl cellulose

NEM:

N-ethylmaleimide

EDTA:

ethylenediaminetetraacetate

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Authors and Affiliations

  1. Department of Biological Sciences, Columbia University, 1305 Altschul Hall, 10027, New York, New York, USA

    Arthur A. Guffanti & W. A. Corpe

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  1. Arthur A. Guffanti
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  2. W. A. Corpe
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Guffanti, A.A., Corpe, W.A. Partial purification and characterization of alpha-glucosidase from Pseudomonas fluorescens W. Arch. Microbiol. 107, 269–276 (1976). https://doi.org/10.1007/BF00425338

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