Nitrogenase activity in cultured Rhizobium sp. strain 32H1

Abstract

Nutritional and physical conditions affecting nitrogenase activity in the strain of “cowpea” rhizobia, 32H1, were examined using cultures grown on agar medium. Arabinose in the basic medium (CS7) could be replaced by ribose, xylose, or glycerol, but mannitol, glucose, sucrose, or galactose only supported low nitrogenase (C₂H₂ reduction) activity. Succinate could be replaced by pyruvate, fumarate, malate, or 2-oxoglutarate, but without any carboxylic acid, nitrogenase activity was low or undetectable unless a high level of arabinose was provided. Inositol was not essential. Several nitrogen sources could replace glutamine including glutamate, urea, (NH₄)₂SO₄ and asparagine. The maximum nitrogenase activity of cultures grown in air at 30°C was observed under assay conditions of pO₂=0.20–0.25 atm and 30°C incubation. Greatest activity occurred after a period of rapid bacterial growth, when viable cell count was relatively constant. Compared with results obtained on the CS7 medium, nitrogenase activity could be substantially increased and/or sustained for longer periods of time by using 12.5 mM succinate and 100 mM arabinose, by increasing phosphate concentration from 2 to 30–50 mM, or by culturing the bacteria at 25°C.