Abstract
Nitrate reduction was studied in the dinoflagellatePeridinium cinctum collected from extensive algal blooms in Lake Kinneret (Israel).
Among several methods tested for the preparation of cell free extracts, only the use of a ground-glass tissue culture homogenizer was found to be efficient. The assimilatory nitrate reductase ofP. cinctum was located in a particulate fraction. In this respect,P. cinctum did not behave like other eukaryotes, such as green algae, but as a prokaryote. Nitrite reductase activity was found in the soluble fraction.
Nitrate reductase used NADH as a preferable electron donor; it reacted also with NADPH but only to give 16.5% of the NADH dependent rate. Methyl viologen and benzyl viologen could also serve as electron donors, with rates higher than the NADH dependent activity (3–6 times and 1.5–3 times, respectively). The Km of nitrate reductase for NADH was 2.8×10−4 M and for NO3-1.9×10−4 M. Flavins did not stimulate the activity, nor was ferricyanide able to activate it. Carboxylic anions stimulated nitrate reductase activity 3–4 fold, an effect which was not mimicked by other anions.
Chlorate, azide and cyanide were competitive inhibitors ofP. cinctum, nitrate reductase withK i values of 1.79×10−3 M, 2.1×10−5 M and 8.9×10−6 M respectively.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Berman T, Pollinger U (1974) Annual and seasonal variations of phytoplankton chlorophyll and photosynthesis in Lake Kinneret. Limnol Oceanogr 19:31–55
Eppley RW, Coatsworth JK, Solorzano L (1969a) Studies of nitrate reductase in marine phytoplankton. Limnol Oceanogr 14:194–201
Eppley RW, Rogers JN, McCarthy JJ (1969b) Half saturation constants for uptake of nitrate and ammonium by marine phytoplankton. Limnol Oceanogr 14:912–920
Gressel J, Berman T, Cohen N (1975) Dinoflagellate ribosomal RNA; an evolutionary relic? J Mol Evol 5:307–313
Guerrero MG, Manzano C, Losada M (1974) Nitrate photoreduction by a cell-free preparation of Anacystis Nidulans. Plant Sci Lett 3:273–278
Harrison WG (1973) Nitrate reductase activity during a dinoflagellate bloom. Limnol Oceanogr 18:456–465
Hartree EF (1972) Determination of protein: A modification of Lowry method that gives a linear photometric response. Anal Biochem 48:422–427
Jetschmann K, Solomonson LP, Vennesland B (1972) Activation of nitrate reductase by oxidation. Biochim Biophys Acta 275:276–278
Loeblich II AR, Klotz LC, Roberts TM, Tuttle RC, Allen JR (1974) The dinoflagellate nucleus. J Phycol (Suppl) 10:14
Losada M, Guerrero MG (1979) The photosynthetic reduction of nitrate and its regulation. In: Barber J (ed) Photosynthesis in relation to model systems. Elsevier, North Holland, pp 365–408
Losada M, Paneque A (1971) In: Colowick SP, Kaplan NO (eds) Methods in enzymology, vol 23. Academic Press, New York, pp 487–494
McCarthy JJ, Wynn D, Bermna T (1982) The uptake of dissolved nitrogenous nutrients in Lake Kinnerel microplankton. Limnol Oceanogr in press
Nicholas DJD, Nason A (1957) Nitrite determination by diazotization and coupling reactions. In: SP Colowick, NO Kaplan (eds) Methods in enzymology, vol III. Academic Press, New York, pp 983–984
Rizzo PJ, Nooden LD (1974) Isolation and partial characterization of dinoflagellate chromatin. Biochim Biophys Acta 349:402–414
Serruya C (1975) Nitrogen and phosphorus balances in Lake Kinneret. Verh Intern Verein Limnol 19:1357–1369
Vennesland G, Guerrero MG (1979) Reduction of nitrate and nitrite. In: M Gibbs, E. Latzko (eds) Encyclopedia of plant physiology. New series, vol 6. Springer, Berlin, pp 425–444
Wynn D, Berman T (1980) Hot water extractable phosphorus. An indicator of nutritional status of peridinium cinctum (Dinophyceae) from Lake Kinneret (Israel). J Ohycol 16:40–46
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hochman, A. Studies of nitrate reductase in the fresh water dinoflagellatePeridinium cinctum . Arch. Microbiol. 133, 62–65 (1982). https://doi.org/10.1007/BF00943771
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00943771