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Projekt Druckansicht

Anaerobic phosphite oxidation - genetic and enzymatic studies

Fachliche Zuordnung Stoffwechselphysiologie, Biochemie und Genetik der Mikroorganismen
Förderung Förderung von 2011 bis 2015
Projektkennung Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 193135621
 
Erstellungsjahr 2015

Zusammenfassung der Projektergebnisse

We find that the energy conservation mechanism during anaerobic phosphite oxidation process in D. phosphitoxidans, based on proteomic, native protein enrichment, enzymatic and genomic data proceeds via substrate phosphorylation where the Adenylylsulfate reductase catalyses the formation of ADP from phosphite and AMP in the presence of an oxidized electron acceptor. The previously described spectrophotometric assay for measurement of phosphite-oxidizing enzyme activity in cell-free extracts of D. phosphitoxidans cells with NAD+ or NADP+ as electron acceptors was improved. A further coupled enzymatic assay was developed, as well as a method for separation and quantitation of the reaction products. All specific proteins expressed in the strain only when phosphite was used as electron donor were cloned, over expressed and partially purified. A protocol for the enrichment of the native protein complex involved in anaerobic phosphite oxidation process was developed. The genome of the strain has been sequenced, assembled, automatically annotated and manually curated. A complete metabolic map and analysis of the genome was performed. Several new metabolic pathways found in the genome were confirmed on physiological level. Most of the proteomic data and the protein enrichment, over-expression and purification data will be published in the future, when the native protein purification protocol and the confirmatory data from the assays will be finalized.

Projektbezogene Publikationen (Auswahl)

 
 

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