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Continuous assay of neuraminidase activity by a bienzyme system immobilized in an artificial membrane
Author(s) -
Kolisis Fragiskos N.,
Thomas Daniel
Publication year - 1987
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.260300204
Subject(s) - neuraminidase , chromatography , chemistry , lactate dehydrogenase , substrate (aquarium) , membrane , enzyme , biochemistry , dehydrogenase , biology , ecology
Lactate dehydrogenase and NANA‐lyase were immobilized in an artificial gelantine membrane. This bienzyme system was used for continuous assay of neuraminidase activity. The K ′ m of the active membrane for lactate dehydrogenase and NANA‐lyase using NADH, pyruvic acid, and N ‐acetylneuraminic acid as substrates were found to be 0.25m M , 0.75m M , and 2.1m M , respectively. The K m of soluble neuraminidase using sialyllactose as substrate was found to be 0.13 m M . The pH optimum for neuraminidase activity was 6.0. At 45°C the reaction rate was higher, and no denaturation phenomena of the immobolized enzymes have been observed. This bienzyme membrane was stable for several weeks stored in the reaction buffer at 4°C.