TY - JOUR
T1 - Evaluation of Alcaligenes eutrophus cells as an NADH regenerating catalyst in organic-aqueous two-phase system
AU - Andersson, Mats
AU - Holmberg, Hans
AU - Adlercreutz, Patrick
PY - 1998/1/5
Y1 - 1998/1/5
N2 - A soluble NAD-dependent hydrogenase contained in Alcaligenes eutrophus was evaluated as a coenzyme regenerating catalyst in an organic-aqueous two- phase (predominantly organic) system. The horse-liver alcohol-dehydrogenase (HLADH) catalyzed reduction of cyclohexanone to cyclohexanol was used as a model reaction. The impact of different solvents (selected to span a large variety of principal properties) on the stability and activity of the HLADH, using substrate-driven regeneration, was studied. Solvents suitable for the HLADH were then selected for an evaluation of the hydrogenase-driven coenzyme regeneration. Hydrophobic solvents such as heptane, toluene; and 1,1,1- trichloroethane were found to be suitable for the coupled reactions catalyzed by HLADH and hydrogenase. Nonimmobilized cells, permeabilized with cetyl- trimethyl-ammonium bromide, were the most efficient preparation for the regeneration of NADH. The use of this preparation in heptane (10% water) was optimized with respect to the yield obtained in the HLADH-catalyzed reduction of cyclohexanone. Using the optimized conditions, yields of 99% cyclohexanol were obtained.
AB - A soluble NAD-dependent hydrogenase contained in Alcaligenes eutrophus was evaluated as a coenzyme regenerating catalyst in an organic-aqueous two- phase (predominantly organic) system. The horse-liver alcohol-dehydrogenase (HLADH) catalyzed reduction of cyclohexanone to cyclohexanol was used as a model reaction. The impact of different solvents (selected to span a large variety of principal properties) on the stability and activity of the HLADH, using substrate-driven regeneration, was studied. Solvents suitable for the HLADH were then selected for an evaluation of the hydrogenase-driven coenzyme regeneration. Hydrophobic solvents such as heptane, toluene; and 1,1,1- trichloroethane were found to be suitable for the coupled reactions catalyzed by HLADH and hydrogenase. Nonimmobilized cells, permeabilized with cetyl- trimethyl-ammonium bromide, were the most efficient preparation for the regeneration of NADH. The use of this preparation in heptane (10% water) was optimized with respect to the yield obtained in the HLADH-catalyzed reduction of cyclohexanone. Using the optimized conditions, yields of 99% cyclohexanol were obtained.
KW - Alcaligenes eutrophus
KW - HLADH
KW - Hydrogenase
KW - NADH regeneration
KW - Organic solvent
UR - http://www.scopus.com/inward/record.url?scp=0032484704&partnerID=8YFLogxK
U2 - 10.1002/(SICI)1097-0290(19980105)57:1<79::AID-BIT10>3.0.CO;2-3
DO - 10.1002/(SICI)1097-0290(19980105)57:1<79::AID-BIT10>3.0.CO;2-3
M3 - Article
C2 - 10099181
AN - SCOPUS:0032484704
SN - 0006-3592
VL - 57
SP - 79
EP - 86
JO - Biotechnology and Bioengineering
JF - Biotechnology and Bioengineering
IS - 1
ER -