Influence of carbohydrates on stability of papain in aqueous tetrahydrofuran mixture

András Szabó, Márta Kotormán, Ilona Laczkó, L. Mária Simon

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Background: The use of enzymes in organic solvents has extended the scale of their practical applications. Papain has been widely used in chemical syntheses because of its broad substrate specificity. The aim of the present study was to improve the stability of papain in aqueous tetrahydrofuran (THF) by using different saccharides. The effects of these carbohydrates on the structure of papain were followed by means of circular dichroism (CD) and fluorescence spectroscopic measurements. Results: In contrast with most organic solvents, 60% (i.e. 600 mL L-1) THF practically inactivated the enzyme within 30 min. Sugars protected papain from THF-induced inactivation in the sequence D-ribose > D-fructose > D-glucose > D-saccharose > D-raffinose. Ribose at 1.6 mol L-1 proved the most effective stabiliser: in 60% THF in the presence of ribose, papain preserved about 55% of its initial activity after 2 h. Fluorescence and near-UV CD spectroscopic measurements revealed local changes in the papain conformation. With decrease in the free amino group content of the enzyme, protein-carbohydrate interactions (Schiff base formation) were detected. Conclusion: These results demonstrate that the catalytic activity and stability of papain may be increased in aqueous THF by using different carbohydrates, when a more compact structure of the enzyme is formed.

Original languageEnglish
Pages (from-to)133-138
Number of pages6
JournalJournal of Chemical Technology and Biotechnology
Volume84
Issue number1
DOIs
Publication statusPublished - Jun 3 2009

Keywords

  • Carbohydrates
  • Schiff base formation
  • Stability of papain
  • Tetrahydrofuran

ASJC Scopus subject areas

  • Biotechnology
  • Chemical Engineering(all)
  • Renewable Energy, Sustainability and the Environment
  • Fuel Technology
  • Waste Management and Disposal
  • Pollution
  • Organic Chemistry
  • Inorganic Chemistry

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