Flexibility of enzymes suspended in organic solvents probed by time-resolved fluorescence anisotropy: Evidence that enzyme activity and enantioselectivity are directly related to enzyme flexibility

Jaap Broos, Antonie J.W.G. Visser, Johan F.J. Engbersen, Willem Verboom, Arie van Hoek, David N. Reinhoudt

Research output: Contribution to journalArticleAcademicpeer-review

114 Citations (Scopus)
76 Downloads (Pure)

Abstract

A time-resolved fluorescence anisotropy study on the molecular flexibility of active-site labeled anthraniloyl--chymotrypsin, dansylsubtilisin Carlsberg, and native subtilisin Carlsberg, suspended in organic solvents, is described. The internal rotational mobility of the fluorophore in the nanosecond time range could be separated from rotation of enzyme aggregates and rapid energy transfer processes. The enzymes suspended in dry organic solvents are less flexible than when dissolved in water. The enzyme flexibility increased with increasing hydration level. The results confirm that the increase in enzyme activity observed upon addition of low amounts of extra water is related to an increase in enzyme flexibility. Differences in enantioselectivity of subtilisin Carlsberg in different organic solvents have been correlated with differences in enzyme flexibility. The relationship between the internal rotational mobility of the fluorophore and the enantioselectivity provides the first experimental evidence that enzyme flexibility and enzyme enantioselectivity are correlated.
Original languageEnglish
Pages (from-to)12657-12663
Number of pages7
JournalJournal of the American Chemical Society
Volume117
Issue number51
DOIs
Publication statusPublished - 1995

Fingerprint Dive into the research topics of 'Flexibility of enzymes suspended in organic solvents probed by time-resolved fluorescence anisotropy: Evidence that enzyme activity and enantioselectivity are directly related to enzyme flexibility'. Together they form a unique fingerprint.

  • Cite this