STUDIES ON DENITRIFICATION:III. ENZYMATIC GAS PRODUCTION BY THE REACTION OF NITRITE WITH HYDROXYLAMINE

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1. When hydroxylamine is added to nitrite in the presence of lactate by using resting cell suspension, both production rate and volume of nitrogen are increased remarkably.<br> 2. Hydroxylamine alone reacts with nitrite and forms a gaseous product, probably nitrous oxide, two nitrogen atoms of which are supposed to originate, one from hydroxylamine and the other from nitrite. This production of gas is inhibited by 10-3M KCN or by a high concentration of hydroxylamine, and the enzyme is destroyed by heating at 100° for 10 minutes. Accordingly there is no other way than to consider that the reaction is enzymatic.<br> 3. Hyponitrite has no effect on production of nitrogen from nitrite; it behaves as an indifferent substance.<br> 4. For NH2OH-NaNO2, system, the Michaelis constants were estimated using cell-free extract. The values are 10-1.2 mole per liter for NH2OH and 10-3.8 mole per liter for NaNO2. Experiments with DMPPD-NaNO2 system indicate that the Michaelis constants are 10-1.5 mole per liter for DMPPD and 10-3.4 mole per liter for NaNO2.<br> 5. It was found that in the DMPPD-NaNO2 system, there is participation by two enzyme components which are separated by a fractionation of the cell-free supernatant obtained by centrifuging at 20, 000×g. using ammonium sulfate.<br> 6. The essential reaction stages in the denitrifying process can be presented in the simple formula<br> The expences for the work were aided partly by the Grant in Aid for the Scientific Research from the Ministry of Education.

Journal

  • J Biochem (Tokyo)

    J Biochem (Tokyo) 45 (2), 133-140, 1958

    The Japanese Biochemical Society

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