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Factor affecting enzyme characteristics of bilirubin oxidase suspensions in organic solvents
Author(s) -
SkrikaAlexopoulos Eleftheria,
Freedman Robert B.
Publication year - 1993
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260410908
Subject(s) - chemistry , aqueous solution , organic solvent , solvent , chromatography , bilirubin oxidase , enzyme , aqueous two phase system , microemulsion , kinetics , enzyme assay , organic chemistry , biochemistry , chemical engineering , pulmonary surfactant , physics , quantum mechanics , engineering
The activity of bilirubin oxidase toward bilirubin was studied in a liquid/solid two‐phase low‐water organic system using a simple spectrophotometric assay to follow the reaction. The enzyme was lyophilized from aqueous solution before being suspended in the organic solvent reaction medium. The activity was significantly influenced by the properties of the aqueous medium from which the enzyme was lyophilized, specifically its pH, and the quantity and nature of the buffering species. Analyses of these effect showed that the role of buffering species in such systems went beyond their effect in fixing the protonation state of the enzyme. The activity was also influenced by the quantity of water added to the organic solvent reaction medium. The reaction was shown to follow Michaelis‐Menten Kinetics, and K m and k cat were determined. The liquid/solid two‐phase system studied was extensively compared to a previously studied water‐in‐oil microemulsion system © 1993 Wiley & Sons, Inc.