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The comparative role of molecular and nitrate-oxygen in the dissimilation of glucose

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Title The comparative role of molecular and nitrate-oxygen in the dissimilation of glucose
Names Bhatt, Rajanikant Premshanker (creator)
Gilmour, C. M. (advisor)
Date Issued 1964-04-03 (iso8601)
Note Graduation date: 1964
Abstract When nitrate is used in a biological electron transport
system as the final acceptor of hydrogen to form water,
the system is characterized as "Nitrate Respiration." However,
the quantitative expression of the over-all participation
of NO₃⁻(0) in the oxidation of glucose by denitrifiers
such as Pseudomonas stutzeri deserves clarification.
In addition, the identity of the pathways of glucose breakdown
that operate with NO₃⁻ are not understood nor is the
dual role of molecular and nitrate oxygen. It is the purpose
of the present study to provide clarification of the
role of nitrate in the utilization of glucose by P. stutzeri.
A strain of P. stutzeri was used throughout the entire
study. The culture was maintained on nitrate broth or
agar. However, a semi-synthetic basal medium was used for
the experimental studies. The individual experiments were carried out using both an electrolytic respirometer and
a microscale radiorespirometric apparatus. Each assembly
provided a closed system thereby allowing for the determination of any evolved gases. Gases were detected using
a Beckman G C-2 chromatograph. Analysis for nitrites,
nitrates and the ammonium ions were determined by established
chemical procedures. Radioactivity of evolved C¹⁴O₂ was determined by counting of barium carbonate planchets
and also by using the Tri-carb Packard Scintillation Spectrometer. Appropriate calculations for specific activity,
specific yield and relative specific activity were
used.
The results of the present study clarify several points
relating to the role of nitrate and molecular oxygen as
terminal hydrogen acceptors in P. stutzeri. First the use
of high nitrate levels (500-1000 ppm NO₃⁻) markedly inhibits
the uptake of molecular oxygen and the evolution of
carbon dioxide. The nitrate inhibition effect progressively
increases with increasing concentrations of nitrate.
However, the observed inhibition decrease with time is at
1000 ppm NO₃⁻ no inhibition is observed after 48 hours
incubation. At relatively low concentrations of nitrate
(250-500 ppm NO₃⁻) both NO₃⁻(O) and molecular (O) act as
dual electron acceptors. In the latter instances, nitrate
contributes one-third and molecular oxygen two-thirds of the total oxygen required for the production of one millimole
of carbon dioxide. A final observation relates to the
pathway of glucose catabolism. High nitrate levels effect
a decrease in the normal phosphogluconate or direct oxidative
route characteristic of pure aerobic respiration in
P. stutzeri. The shift in the glucose breakdown pathway
appears to go over to classical glycolysis. Thus, "nitrate
respiration" in P. stutzeri appears to make use of the
EMP pathway rather than the route to pentose formation.
This aspect of the study is presently being given additional
emphasis.
Genre Thesis/Dissertation
Topic Pseudomonas
Identifier http://hdl.handle.net/1957/48175

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