PMID: 6988213Jan 1, 1980Paper

A simple procedure for covalent immobilization of NADH in a soluble and enzymically active form

European Journal of Biochemistry
C W FullerH J Bright

Abstract

Using a new and simplified technique for covalent immobilization of adenine nucleotides [Fuller, C. W. and Bright, H. J., J. Biol. Chem. 252, 6631 (1977)], we have prepared immobilized NADH using two water-soluble polymers and tested these preparations for activity with several dehydrogenases. The first polymer used, having a weight-average molecular weight of about 350000, is a copolymer of methacrylyl choline and the epoxide-containing monomer 3-[4-(2,3-epoxypropoxy)butoxy]-2-hydroxypropyl acrylate. Immobilization of NADH onto this copolymer was accomplished in three steps, namely, alkylation of NAD at N-1, reduction of the nicotinamide moiety with dithionite and Dimroth rearrangement of the alkyl linkage from the N-1 to the C-6 amino position. The second copolymer tested was a copolymer of the same epoxide-containing monomer and N-methacrylyl-2-glucosamine. Using this copolymer, immobilization of NADH through the adenine C-6 amino position was accomplished in a single step. Measurements of the steady-state kinetics of five dehydrogenases at pH 7 and pH 9 showed that, on the average, V and Km values obtained with the immobilized NADH were, respectively, about one third and twice those found for the free coenzyme. In general, ...Continue Reading

References

Jul 1, 1976·Analytical Chemistry·W J Blaedel, R A Jenkins
Jun 1, 1975·Archives of Biochemistry and Biophysics· Lee C-Y, N O Kaplan
Jun 1, 1975·European Journal of Biochemistry·P ZappelliL Re
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Dec 29, 1974·Biochimica Et Biophysica Acta·P Davies, K Mosbach
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Oct 1, 1968·Biochemistry·J B Macon, R Wolfenden
May 1, 1967·Journal of Chromatography·K A Granath, B E Kvist

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Citations

Mar 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·M O MånssonK Mosbach
Apr 27, 2007·Biotechnology Advances·Wenfang Liu, Ping Wang

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