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Enzymatic activity and stability of d ‐fructose dehydrogenase and sarcosine dehydrogenase immobilizd onto giant vesicles
Author(s) -
Kato Keiichi,
Walde Peter,
Mitsui Hirokazu,
Higashi Nobuyasu
Publication year - 2003
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.10784
Subject(s) - dehydrogenase , vesicle , chemistry , biochemistry , enzyme , sorbitol dehydrogenase , phosphatidylcholine , sarcosine , chromatography , immobilized enzyme , fructose , membrane , phospholipid , glycine , amino acid
Stable vesicles with diameters between about 1 and 10 μm 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°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. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 84: 415–423, 2003.