Abstract
The Fe protein ofAnabaena cylindrica was first separated and purified by chromatography through DEAE-cellulose columns then by gel electrophoresis. The specific activity was up to 142.46 nmol C2H4/mg protein · min. It was homogeneous as shown by 1) a single band in the gel electrophorogram; 2) absence of Mo and tryptophan; 3) content of about 3.4 atoms of Fe per mole protein. The molecular weight of the Fe protein ofA. cylindrica was about 61,000 daltons as estimated by SDS-gel electrophoresis and calculated from the amino acid composition. The residues of aspartate and glutamate were about 2.6 times that of arginine and lysine in the Fe protein. Crossing Fe protein ofA. cylindrica with Mo−Fe protein ofAzotobacter vinelandii gave positive result. The reciprocal crossing also showed activity.
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References
Bradford, M. M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Analytical Biochemistry 72 (1–2): 248–254.
Davis, B. J., 1964. Disc electrophoresis. II. Method and application to human serum proteins.Annals. N.Y. Acad. Sci. 121: 404.
Eady, R.R., B.E. Smith, K.A. Cook et al., 1972. Nitrogenase ofKlebsiella pneumoniae. Purification and properties of the component proteins.Biochem. J. 128: 655–675.
Kelly, M., R. V. Klucas and R.H. Burris, 1967. Fractionation and storage of nitrogenase fromAzotobacter vinelandii.Biochem. J. 105 (1): 3C-4C.
Nakamura, M., 1980. Biological Nitrogen Fixation. Tokyo Academic Centre of Publication, Tokyo, 111 pp. (in Japanese).
Oppenheim, J. and L. Marcus, 1970. Correlation of ultrastructure inAzotobacter vinelandii with nitrogen source for growth.J. Bacteriol. 101 (1): 286–291.
Shah, V.K. and W.J. Brill, 1973. Nitrogenase. IV. Simple method of purification to homogeneity of nitrogenase components fromAzotobacter vinelandii.Biochem. Biophys. Acta 305 (2): 445–454.
Thoruber, J.P. and J.M. Olson, 1968. The chemical composition of a crystalline bacteriochlorophyll protein complex isolated from the green bacterium,Chloropscudomonas ethylicym.Biochemistry 7 (6): 2242–2249.
Vandecasteele, J.P. & R.H. Burris, 1970. Purification and properties of the contituents of the nitrogenase complex from Clostridium pasteurianum.J. Bacteriol. 101: 794–801.
Weber, K. and M. Osborn, 1969. The reliability of molecular weight determinations by dodecyl sulfate polyacrylamide gel electrophoresis.J. Biochem. 244: 4406
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Daixian, D., Huimin, L., Zhenrong, H. et al. Purification and some properties of Fe protein of nitrogenase from. Chin. J. Ocean. Limnol. 8, 378–384 (1990). https://doi.org/10.1007/BF02849684
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DOI: https://doi.org/10.1007/BF02849684