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Optimized nitrate reductase assay predicts the rate of nitrate utilization in the halotolerant microalga Dunaliella viridis

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Abstract

An in situ method for measuring nitrate reductase (NR) activity in Dunaliella viridis was optimized in terms of incubation time, concentration of KNO3, permeabilisers (1-propanol and toluene), pH, salinity, and reducing power (glucose and NADH). NR activity was measured by following nitrite production and was best assayed with 50 mM KNO3, 1.2 mM NADH, 5% 1-propanol (v/v), at pH 8.5. The estimated half-saturation constant (Ks) for KNO3 was 5 mM. Glucose had no effect as external reducing power source, and NADH concentrations >1.2 mM inhibited NR activity. Nitrite production was linear up to 20 min; longer incubation did not lead to higher nitrate reduction. The use of the optimized assay predicted the rate of NO −3 removal from the external medium by D. viridis with high degree of precision.

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Authors and Affiliations

  1. Departamento de Ecología, Facultad of Ciencias, Universidad de Málaga, 29071, Málaga, Spain

    F. Javier, L. Gordillo, Carlos Jiménez & F. Xavier Niell

  2. Departamento de Ecología, Facultad de Ciencias del Mar, Universidad de Cádiz, Cádiz, Spain

    Alfonso Corzo

Authors
  1. F. Javier
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  2. L. Gordillo
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  3. Carlos Jiménez
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  4. Alfonso Corzo
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  5. F. Xavier Niell
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Correspondence to Carlos Jiménez.

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Javier, F., Gordillo, L., Jiménez, C. et al. Optimized nitrate reductase assay predicts the rate of nitrate utilization in the halotolerant microalga Dunaliella viridis. Journal of Applied Phycology 9, 99–106 (1997). https://doi.org/10.1023/A:1007978512458

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