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Preparation and properties of immobilized flavokinase
Author(s) -
Merrill Alfred H.,
McCormick Donald B.
Publication year - 1979
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.260210909
Subject(s) - chemistry , chromatography , biochemical engineering , engineering
Flavokinase (ATP: riboflavin 5'‐phosphotransferase, EC 2.7.1.26) purified from rat liver by affinity chromatography, has been immobilized by amide linkage to aminoalkyl‐agarose beads. The immobilized enzyme differs from the soluble enzyme in having greater stability slightly higher K m for the substrates, riboflavin and ATP, a broader pH optimum, and a lower energy of activation. These results suggest that the immobilized enzyme is influenced by the microenvironment of the bead and is subject to some degree of internal diffusional limitation. A Small (3ml), continuous, plug‐flow reactor prepared with immobilized flavokinase effects 5% conversion of riboflavin to riboflavin 5′‐phosphate (FMN) with a flow rate of 0.16 ml/min, which corresponds to an output of 5 nmol FMN/min. Immobilized flavokinase is effective for phosphorylating riboflavin and numerous riboflavin analogs and provides a facile methods for preparing exclusively, unlike other synthetic methods, the 5′‐phosphates.