Detection of an insensitive ornithine‐carbamoyltransferase in strains of <i>Pseudomonas syringae</i> pv. <i>phaseolicola</i> with different phytotoxin‐generating capacities

Jahn, O.; Sauerstein, J.; Laplace, Frank; Reuter, G.

doi:10.1002/jobm.3620290517

Cited by 3 publications

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“…The OCT from E. roli N 100 and Phaseoius vulgaris L., however, showed only sensitive reaction ( JAHN 1984, JAHN et al 1989. The fact that the OCT in cell-free extracts from P. syringae pv.…”

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“…phaseolicola (YOUNG et al 1978), the producer of phaseolotoxin and PNSOrn ( MOORE et al 1984), it would seem that an insensitive target enzyme is a part of the mechanism for self-protection : We have detected two particular ornithine carbamoyltransferases which differ in their sensitivity to phaseolotoxin and PNSOrn, in cells of P. syringae pv. phaseolicola (JAHN et al 1985(JAHN et al , 1987(JAHN et al , 1989. We isolated and.described a sensitive (OCT I ) and an insensitive enzyme (OCT 2).…”

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Inhibition of ornithine carbamoyltransferase from Pseudomonas syringae pv. syringae W50 by phaseolotoxin

Jahn

Sauerstein

Reuter

1989

J. Basic Microbiol.

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In contrast to the producer of phaseolotoxin and Orn-P(O)(NH2)-NH-SO3H (PNSOrn), Pseudomonas syringae pv. phaseolicola, which possesses a sensitive and an insensitive type of ornithine carbamoyltransferase (OCT, E.C. 2.1.3.3.), in Pseudomonas syringae pv. syringae W50, an organism which produce neither phaseolotoxin nor PNSOrn, only one type of OCT could be detected. This enzyme is highly sensitive to phaseolotoxin. This result supports our hypothesis that the existence of an insensitive ornithine carbamoyltransferase is an important prerequisite for the synthesis of phaseolotoxin in P. syringae pv. phaseolicola and that this enzyme does not occur generally in P. syringae spec.

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Inhibition of ornithine carbamoyltransferase from Pseudomonas syringae pv. syringae W50 by phaseolotoxin

Jahn

Sauerstein

Reuter

1989

J. Basic Microbiol.

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Biosynthesis of homoarginine and ornithine as precursors of the phytoeffector phaseolotoxin by the amidinotransfer from arginine to lysine catalyzed by an amidinotransferase in Pseudomonas syringae pv. phaseolicola

Märkisch

Reuter

1990

J. Basic Microbiol.

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(Friedrich-Schiller-Universitiit Jena, Sektion Biologie --M ikrobielle Biochemie)Biosynthesis of homoarginine and ornithine as precursors of the phytoeffector phaseolotoxin by the amidinotransfer from arginine to lysine catalyzed by an amidinotransferase in Pseudomonas syringae pv. phaseolicola U. MARKISCH and G. REUTER f Receiuetl 16 March lY8YIAccepfed 1 DecemherlY89)In the phytopathogenic Pseudomonos syrinycre pv. phaseolicola an amidinotransferase was detected and characterized which catalyzes the amidinotransfer from arginine to lysine producing the phaseolotoxin precursors homoarginine and ornithine. The enzyme was purified 419-fold by affinity chromatography on homoarginine-Sepharose. It had an apparent mol. wt. of 200000 and exhibited MICHAELIS-MENTEN kinetics with K,,, 7.7 mM for arginine and K,,, 4.2 mM for lysine. Ornithine competitively inhibited the enzyme,with a Ki of 0.8 mM. During cultivation of the pseudomonad the highest enzyme activity corresponds with the highest production rate of phaseolotoxin. At low toxin production, mutants exhibit a low enzyme activity and vice versa. Only those strains of different pathovarieties which are able to produce phaseolotoxin contain the enzyme. At 30 "C cultivation temperature, the enzyme activity was 50 7; of that at 18 "C, and toxin production decreased with increasing temperature. When microbes were grown on ribose, their enzyme activity was 60',', lower than grown on glucose or glycerol. Toxin production was decreased to about 10 "/d when growing on ribose. Amidinotransferase is considered to be one of the key enzymes of phaseolotoxin biosynthesis.Pseudonzonas syringae pv. phaseolicola ( P . phaseolicola) causes the halo blight disease in bean, Pltaseouls vulgaris (RUDOLPH 1976, DURBIN 1985. It produces phaseolotoxin, a tripeptide consisting of homoarginine, alanine and ornithine which is substituted in its b-aminogroup, om-P(0) (NHJ-NH-S0,H. Furthermore it produces this substituted ornithine (PNSOrn) 1977, PATIL et al. 1970, TAM and PATIL 1972. Searching for the molecular basis of the resistance of P. phaseolicola against the selfproduced toxin we found a second OCT, which is insensitive to the toxin (JAHN el at. 1985(JAHN el at. , 1987(JAHN el at. , 1989a(JAHN el at. , 1989b. It is suggested that PNSOrn is the decisive ingredient of phaseolotoxin and that alanyl-homoarginine functions as a toxin vehicle in its transport to the plant cell (MITCHELL 1976).Until now nothing was known about the in vitro biosynthesis of phaseolotoxin and its precursor homoarginine. Arginine is wellknown as a common amidine donor for guanidines. So we did expect that homoarginine and ornithine, two of the phaseolotoxin precursors, were prepared by a transamidination reation from lysine and arginine. The aim of this study was to find out and to characterize an amidinotfansferase in cell free extracts of P. phaseolicola, to rnofify methods for the purification of this enzyme opening possibilities for its additional characterization, and to test the conncection between enz...

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Current Ideas on the Genetics and Regulation of the Synthesis of Phaseolotoxin in Pseudomonas Syringae PV. Phaseolicola

Alvarez‐Morales¹,

López‐López²,

Hernández-Flores³

2004

Pseudomonas

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Detection of an insensitive ornithine‐carbamoyltransferase in strains of Pseudomonas syringae pv. phaseolicola with different phytotoxin‐generating capacities

Cited by 3 publications

References 7 publications

Inhibition of ornithine carbamoyltransferase from Pseudomonas syringae pv. syringae W50 by phaseolotoxin

Inhibition of ornithine carbamoyltransferase from Pseudomonas syringae pv. syringae W50 by phaseolotoxin

Biosynthesis of homoarginine and ornithine as precursors of the phytoeffector phaseolotoxin by the amidinotransfer from arginine to lysine catalyzed by an amidinotransferase in Pseudomonas syringae pv. phaseolicola

Current Ideas on the Genetics and Regulation of the Synthesis of Phaseolotoxin in Pseudomonas Syringae PV. Phaseolicola

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