Deactivation of formate dehydrogenase (FDH) in solution and at gas-liquid interfaces.
Enzymes, increasingly important in the synthesis of fine chemicals and pharmaceutical intermediates, are often insufficiently stable under reacting conditions. We have investigated the stability, in homogeneous aqueous solution and at gas-liquid interfaces, of formate dehydrogenase (FDH), important for cofactor regeneration, from Candida boidinii and overexpressed in E. coli. When exposed to mechanical stress, residual activity, [E](t)/[E](0), and residual protein were found to scale proportionally with gas-liquid surface area in the bubble column, verifying a surface-driven process, and with time and total throughput in a gear pump, but did not seem to be influenced much by shear in a Couette viscometer. All FDH variants are deactivated by chaotropes but not kosmotropes: the first-order deactivation constant k(d) correlates well with the Jones-Dole coefficient B but not well with the surface tension increment deltasigma of various concentrated ammonium salt solutions. This finding might provide guidance for focusing the search for quantitative theories of Hofmeister effects.
Bommarius
Andreas S
AS
Research, Development and Appl. Technology Fine Chemicals, Degussa AG, P.O. Box 1345, D-63403 Hanau, Germany. andreas.bommarius@chbe.gatech.edu
Karau
Andreas
A
D019149:Bioreactors; D001709:Biotechnology; D002175:Candida / Q000201:enzymology / Q000235:genetics; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology; D005427:Flocculation; D005560:Formate Dehydrogenases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D005740:Gases; D014644:Genetic Variation; D007700:Kinetics; D009243:NAD / Q000378:metabolism; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D012492:Salts; D012996:Solutions; D013314:Stress, Mechanical; D014783:Viscosity
D005740:Gases; D011994:Recombinant Proteins; D012492:Salts; D012996:Solutions; D009243:NAD; D005560:Formate Dehydrogenases