Physiological genetics of denitrification: A route to conserving fixed nitrogen: Progress (report), January 1986-January 1987

Technical Report · Thu Jan 01 04:00:00 EST 1987

OSTI ID:7092409

Ingraham, J L

A set of 35 mutant strains of Pseudomonas stutzeri blocked in the final step of the denitrification pathway, have been analyzed. These strains, isolated following treatment with a frame-shift mutagen grow anaerobically at near normal rates when nitrate or nitrite ions are present as the only reductant but fail to grow, or grow poorly, if N/sub 2/O is the only reductant. Analyzing these strains using two dimensional gels to examine the proteins they produce revealed that the strains can be grouped into classes that do not produce protein a (a/sup -/b/sup +/); those that do not produce protein b (a/sup +/b/sup -/) or produce neither protein a nor b (a/sup -/b/sup -/); and those that do not produce protein a but instead produce a new protein, e, that migrates to a position on the gel slightly above and to the right of protein b. Protein b has been identified as the copper-containing N/sub 2/O reductase described by Zumft. Protein a, which our mutant studies have identified and shown also to be essential for the reduction of N/sub 2/O, is membrane-bound. Like N/sub 2/O reductase, protein a also contains copper. As judged by the intensity of spots on two dimensional gels we found that the amount of protein a produced by cells grown anaerobically in complex medium in the presence of N/sub 2/O increases with the amount of copper in the medium. The protein seems to be copper-inducible. 1 fig., 1 tab.

• Research Organization: California Univ., San Diego, La Jolla (USA)
• DOE Contract Number: FG03-85ER13356
• OSTI ID: 7092409
• Report Number(s): DOE/ER/13356-T1; ON: DE87006426
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES
AUTORADIOGRAPHY
BACTERIA
BIOLOGICAL PATHWAYS
CARBON 14 COMPOUNDS
CHEMICAL REACTIONS
CLONING
COPPER
DENITRIFICATION
DNA-CLONING
ELECTROPHORESIS
ELEMENTS
ENZYME INDUCTION
ENZYMES
FRACTIONATION
GENE REGULATION
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MEMBRANE TRANSPORT
METALS
MICROORGANISMS
MUTANTS
NITRO-GROUP DEHYDROGENASES
NITROREDUCTASES
OXIDOREDUCTASES
PSEUDOMONAS
SEPARATION PROCESSES
TRACER TECHNIQUES
TRANSITION ELEMENTS