Enzymatic activity and stability of D-fructose dehydrogenase and sarcosine dehydrogenase immobilized onto giant vesicles.
Stable vesicles with diameters between about 1 and 10 mum were prepared by a particular emulsification technology that involved the use of the surfactants Span 80 and Tween 80 and the phospholipid lecithin (phosphatidylcholine from soybeans). Two membrane enzymes, d-fructose dehydrogenase from Gluconobacter sp. (FDH) and sarcosine dehydrogenase from Pseudomonas putida (SDH), were for the first time immobilized onto the bilayer membranes of these type of vesicles; and the catalytic activity and enzymatic stability were measured and compared with the enzymes in a vesicle-free solution. The enzyme activity as well as stability considerably increased upon immobilization. In particular, immobilized FDH at 25 degrees C was stable for at least 20 days, while the activity of the free enzyme dropped to about 20% of its initial value during the same period of time. In contrast to FDH and SDH, immobilization of sorbitol dehydrogenase from Gluconobacter suboxydans (SODH) was not successful, as no improved activity or stability could be obtained.
Kato
Keiichi
K
Department of Applied Chemistry, Faculty of Engineering, Ehime University, Bunkyo chyo 3, Matsuyama, 790 Japan. kato@eng.ehime-u.ac.jp
Walde
Peter
P
Mitsui
Hirokazu
H
Higashi
Nobuyasu
N
D002237:Carbohydrate Dehydrogenases / Q000737:chemistry*; D020099:Coated Materials, Biocompatible; D004789:Enzyme Activation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry; D006601:Hexoses / Q000737:chemistry; D007064:L-Iditol 2-Dehydrogenase / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D008567:Membranes, Artificial; D010089:Oxidoreductases, N-Demethylating / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry; D011136:Polysorbates; D050892:Sarcosine Dehydrogenase; D013379:Substrate Specificity
D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D006601:Hexoses; D008051:Lipid Bilayers; D008081:Liposomes; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D011136:Polysorbates; C018665:sorbitan monooleate; D002237:Carbohydrate Dehydrogenases; D007064:L-Iditol 2-Dehydrogenase; C028761:D-fructose 5-dehydrogenase; D010089:Oxidoreductases, N-Demethylating; D050892:Sarcosine Dehydrogenase