PMID: 1204624Nov 15, 1975Paper

Properties of immobilized flavodoxin from Peptostreptococcus elsdenii. An affinity ligand for the purification of riboflavin 5'-phosphate (FMN) and its analogues

European Journal of Biochemistry
S G Mayhew, M J Strating

Abstract

The small flavoprotein, flavodoxin, isolated from Peptostreptococcus elsdenii, has been covalently coupled to CNBr-activated Sepharose 4B. The immobilized protein replaces ferredoxin as an electron carrier in hydrogen production from dithionite or pyruvate in the presence of ferredoxin-free extracts of P. elsdenii; compared with soluble flavodoxin, its activities in these systems are 13% and 3.5% respectively. Acid treatment reversibly dissociates FMN from the immobilized protein. The dissociation constant of the complex with FMN, determined by fluorimetric titration, is 1.5 (+/- 0.4) nM, and is therefore very little different from that of soluble flavodoxin. Like soluble apoflavodoxin, the immobilized apoprotein is highly specific for flavins with an N-10 side-chain of 5 carbon atoms and a C-5' phosphate group. Approximately half of the flavin impurity in commercial preparations of FMN (12-15% of the total flavin), and similar impurity in synthetic analogues of FMN, is not separated by conventional purification procedures, but it is readily and conveniently removed by affinity chromatography with apoflavodoxin as the immobilized ligand. The immobilized protein is stable for long periods; its capacity for FMN decreases by only ...Continue Reading

References

May 12, 1971·Biochimica Et Biophysica Acta·S G Mayhew
Jun 1, 1948·The Journal of Experimental Medicine·E S G BARRONZ B MILLER

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Citations

Sep 1, 1979·Biotechnology and Bioengineering·A H Merrill, D B McCormick
Sep 19, 2008·Proceedings of the National Academy of Sciences of the United States of America·Joseph A Cotruvo, JoAnne Stubbe
Nov 1, 1969·Proceedings of the National Academy of Sciences of the United States of America·J R BenemannD I Arnon
Nov 1, 1972·Proceedings of the National Academy of Sciences of the United States of America·K D WatenpaughM Dubourdieu
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Dec 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·S GhislaJ M Lhoste
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Jul 27, 2012·The Journal of Biological Chemistry·Pilar Bernal-BayardJosé A Navarro
Jul 11, 2007·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Julie WolfováIvana Kutá Smatanová
Oct 1, 1976·European Journal of Biochemistry·G Scola-NagelschneiderP Hemmerich
Mar 15, 1979·European Journal of Biochemistry·H FennerV Massey
Apr 5, 1983·European Journal of Biochemistry·Y S Choong, V Massey
Jun 10, 1980·Biochemistry·S GhislaT P Singer
Oct 16, 1978·Biochemical and Biophysical Research Communications·K Schneider, H G Schlegel
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Aug 21, 2002·The Journal of Biological Chemistry·Sumita Chakraborty, Vincent Massey
Dec 15, 1981·Biochimica Et Biophysica Acta·P Nielsen, A Bacher
Oct 1, 1978·Archives of Biochemistry and Biophysics·G Bookjans, P Böger

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