Purification and partial characterization of three forms of α-glucosidase from the fruit fly Drosophila melanogaster

Teiichi Tanimura, Keisuke Kitamlira, Tomoko Fukuda, Toshihide Kikuchi

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Abstract

Three forms of β-glucosidase, I, II, and Ill, have been purified from the whole body extract of adult flies of Drosophila melanogaster in yields of 2·1, 5·3, and 6·7%, respectively. The purification procedures involved ammonium sulfate fractionation, Con A-Sepharose 4B affinity chromatography, DEAE-Sepharose CL-6B ion exchange chromatography, Sephacryl S-200 gel filtration, and preparative gel electrophoresis. Each purified enzyme showed a single band on polyacrylamide gel on both protein and enzyme activity staining. The molecular weights of α-glucosidases I, II, and liii were estimated to be 200,000, 56,000, and 76,000, respectively, by gel filtration. SDS gels indicated that α-glucosidases II and III were each composed of a single polypeptide chain, whereas α-glucosidase I was composed of two identical subunits. Both α-glucosidases II and III hydrolyzed sucrose and p-nitrophenyl-α-D-glucoside (PNPG), but α-glucosidase I hydrolyzed PNPG to a much lesser extent than sucrose. For sucrose the pH optima of α-glucosidases I, II, and III were pH 6·0, 5·0, and 6·0 and the Km values were 13·1, 8·9, and 10 nM, respectively. For PNPG the pH optima of a-glucosidases II and III were pH 5·5 and 6·5 and the Km values were 0·77 and 0·21 mM respectively.

Original languageEnglish
Pages (from-to)123-130
Number of pages8
JournalJournal of biochemistry
Volume85
Issue number1
DOIs
Publication statusPublished - Jan 1 1979

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Glucosidases
Fruits
Drosophila melanogaster
Diptera
Purification
Fruit
Glucosides
Gels
Sucrose
Gel Chromatography
Affinity chromatography
Ammonium Sulfate
Enzyme activity
Fractionation
Chromatography
Electrophoresis
Sepharose
Ion Exchange Chromatography
Enzymes
Ion exchange

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Purification and partial characterization of three forms of α-glucosidase from the fruit fly Drosophila melanogaster. / Tanimura, Teiichi; Kitamlira, Keisuke; Fukuda, Tomoko; Kikuchi, Toshihide.

In: Journal of biochemistry, Vol. 85, No. 1, 01.01.1979, p. 123-130.

Research output: Contribution to journalArticle

Tanimura, Teiichi ; Kitamlira, Keisuke ; Fukuda, Tomoko ; Kikuchi, Toshihide. / Purification and partial characterization of three forms of α-glucosidase from the fruit fly Drosophila melanogaster. In: Journal of biochemistry. 1979 ; Vol. 85, No. 1. pp. 123-130.
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