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Title: The impact of nitrite on aerobic growth of Paracoccus denitrificans PD1222
Author: Hartop, Katherine
ISNI:       0000 0004 5367 1414
Awarding Body: University of East Anglia
Current Institution: University of East Anglia
Date of Award: 2014
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The effect of nitrite stress induced in Paracoccus denitrificans PD1222 was examined using additions of sodium nitrite to an aerobic bacterial culture. Nitrite generates a strong stress response in P. denitrificans, causing growth inhibition. This is dependent on both the concentration of nitrite present and the pH. The pH dependent effect of nitrite growth inhibition is likely a result of nitrite and free nitrous acid (FNA; pKa = 3.16) and subsequent reactive nitrogen oxides, generated from the intracellular passage of FNA into P. denitrificans. A flavohemoglobin (fhb; Pd1689) and its associated NsrR family, transcriptional regulator (Pd1690), were transcribed above a ≥2 fold expression filter (p ≤ 0.05) at 95% significance in qRT-PCR and a type II microarray transcriptional analysis at 12.5 mM nitrite in batch culture. Additionally, >25 fold expression of the flavohemoglobin was confirmed by qRT-PCR in continuous culture with nitrite. A deletion mutant determined fhb to be involved in conveying nitrite resistance at high nitrite concentrations and is linked to a stimulation of biomass generated by the presence of nitrite. The cytochrome ba3 oxidase was found to be associated with nitrite in transcriptional analysis, suggesting the uncoupling of the protonmotive force caused by the transport of FNA across the membrane, and subsequent dissociation in the cytoplasm were reduced by a method of counterbalance. No nitrate accumulation was seen and nitrous oxide levels were above that observed for atmospheric background levels. The microarray analysis was used to confirm that in batch growth at 12.5 mM nitrite addition, P. denitrificans shows an overall stress response associated with protein, DNA and lipid repair, with the addition of fhb detoxification and action of the cytochrome ba3 oxidase. It is therefore suggested that nitrite presents a pHdependent stress response in P. denitrificans, likely due to the production of associated reactive nitrogen species such as NO from the internalisation of FNA and the uncoupling of the protonmotive force.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available