6-Phosphogluconate dehydratase deficiency in pleiotropic carbohydrate-negative mutant strains of Pseudomonas aeruginosa

Blevins, W. T.; Feary, Thomas W.; Phibbs, Paul V.

doi:10.1128/jb.121.3.942-949.1975

Cited by 32 publications

(26 citation statements)

References 28 publications

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“…It was early suggested that glycerol catabolism relies on a functional and active ED pathway in P. aeruginosa (Blevins et al, 1975;Heath and Gaudy, 1978) and in P. putida (Aparicio et al, 1971;Vicente and Cánovas, 1973). Heath and Gaudy (1978) proposed that glycerol metabolism depends on the activation of GAPDH and, therefore, on an active metabolism of the hexosephosphate derivatives.…”

Section: Discussionmentioning

confidence: 99%

The role of GlpR repressor in Pseudomonas putida KT2440 growth and PHA production from glycerol

Escapa

Cerro

Garcı́a

et al. 2012

Environmental Microbiology

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Pseudomonas putida KT2440 has evolved a tightly regulated system for metabolizing glycerol implying a prolonged growth lag-phase. We have learnt that this fact can be avoided by the addition of small amounts of some growth precursors. The addition of 1 mM octanoic acid as co-feeder completely eliminated the lag-phase, resulting in an improvement, in terms of invested time, of both growth and polyhydroxyalkanoates (PHA) accumulation. To investigate this phenomenon, we have followed co-metabolic approaches combined with mutations of the specific and global regulatory networks that connect glycerol catabolism and PHA synthesis. By using mutant strains in metabolic genes from the PHA and tricarboxylic acid (TCA) cycles, we have demonstrated that the co-feeding effect is independent of PHA accumulation, but driven on active glyoxylate shunt and Entner-Doudoroff (ED) routes. These findings suggested that the effect of octanoate on glycerol metabolism could rely, either on a global activation of the cell energy state, or on the generation of specific metabolites or cofactors needed for the activation of glycerol metabolism. Our results addressed GlpR as the key factor controlling the efficient utilization of glycerol as growth precursor in P. putida KT2440. Accordingly, a glpR knockout mutant of P. putida KT2440 showed an elimination of the lag-phase when cultured on glycerol in the absence of co-feeder. Besides, the production of PHA in this strain was increased near twofold, resulting in a higher final yield in terms of PHA accumulation. The repressor activity of the GlpR protein over the glp genes in the absence of glycerol was finally demonstrated by qRT-PCR. This work contributed to unravel the physiological causes of the long lag-phase produced by glycerol in the model strain P. putida KT2440 that hinders its use as carbon source in biotechnological applications for generating valuable products.

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Section: Discussionmentioning

confidence: 99%

The role of GlpR repressor in Pseudomonas putida KT2440 growth and PHA production from glycerol

Escapa

Cerro

Garcı́a

et al. 2012

Environmental Microbiology

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“…Since the assay depends upon the conversion of NADPf into NADPH, absolute values of enzyme activity should be regarded with caution. In a study of carbohydrate catabolism in mutant strains of P. aeruginosa (Blevins et al, 1975), increased 6-phosphogluconate dehydrogenase activity was detected only in those strains containing elevated levels of Entner-Doudoroff enzymes. The generation of glyceraldehyde 3-phosphate by this pathway may provide substrate for endogenous glyceraldehyde-3-phosphate dehydrogenase, thus creating errors in the assay.…”

Section: R E S U L T Smentioning

confidence: 99%

Glucose Metabolism in Xanthomonas campestris

Whitfield¹,

Sutherland²

1982

Microbiology

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~ ~~ ~ ~~The pathways of glucose metabolism in Xanthomonas campestris closely resemble those described for Pseudomonas aeruginosa, in which two discrete systems exist for the uptake of glucose, one periplasmic and oxidative requiring glucose dehydrogenase activity, the second intracellular and phosphorylative. These systems lead to the production of 6phosphogluconate, which may be metabolized via the Entner-Doudoroff pathway and the pentose phosphate pathway. Terminal oxidation is mediated by the tricarboxylic acid cycle.A mucoid strain of X . campestris lacked glucose dehydrogenase activity and thus the oxidative pathway was not functional; the pentose phosphate pathway was also inoperative in this strain, as evidenced by the absence of 6-phosphogluconate dehydrogenase activity. Nonmucoid mutants appeared identical to the parent strain; however, a second group of mutants (crenated mutants) were derepressed for both glucose dehydrogenase and 6phosphogluconate dehydrogenase. Gluconate kinase was not detected in any of the Present address:

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“…Longer incubation led to a faint growth on fructose plates. Mutants lacking the dehydrase and the aldolase of the Entner-Doudoroff pathway were impaired iti growth on most substrates, the dehydrase mutant being glycerol positive (2).…”

Section: Synthesismentioning

confidence: 99%

“…Glycerol metabolism also culminates in the production of glyceraldehyde 3-phosphate as depicted in Fig. 5 through glycerol kinase, aglycerophosphate dehydrogenase, and triose phosphate isomerase (2). It would be interesting to examine the behavior of mutants lacking fructose diphosphatase and fructose diphosphate aldolase.…”

Section: Downloaded Frommentioning

confidence: 99%

Alginic acid synthesis in Pseudomonas aeruginosa mutants defective in carbohydrate metabolism

Banerjee¹,

Vanags²,

Chakrabarty³

et al. 1983

J Bacteriol

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Mutant cells of mucoid Pseudomonas aeruginosa isolated from cystic fibrosis patients were examined for their ability to synthesize alginic acid in resting cell suspensions. Unlike the wild-type strain which synthesizes alginic acid from glycerol, fructose, mannitol, glucose, gluconate, glutamate, or succinate, mutants lacking specific enzymes of carbohydrate metabolism are uniquely impaired. A phosphoglucose isomerase mutant did not synthesize the polysaccharide from mannitol, nor did a glucose 6-phosphate dehydrogenase mutant synthesize the polysaccharide from mannitol or glucose. Mutants lacking the Entner-Doudoroff pathway dehydrase or aldolase failed to produce alginate from mannitol, glucose, or gluconate, as a 3-phosphoglycerate kinase or glyceraldehyde 3-phosphate dehydrogenase mutant failed to produce from glutamate or succinate. These results demonstrate the primary role of the Entner-Doudoroff pathway enzymes in the synthesis of alginate from glucose, mannitol, or gluconate and the role of glyceraldehyde 3-phosphate dehydrogenase reaction for the synthesis from gluconeogenic precursors such as glutamate. The virtual absence of any activity of phosphomannose isomerase in cell extracts of several independent mucoid bacteria and the impairment of alginate synthesis from mannitol in mutants lacking phosphoglucose isomerase or glucose 6-phosphate dehydrogenase rule out free mannose 6-phosphate as an intermediate in alginate biosynthesis. grown overnight in the synthetic medium described by 238 on August 4, 2020 by guest

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6-Phosphogluconate dehydratase deficiency in pleiotropic carbohydrate-negative mutant strains of Pseudomonas aeruginosa

Cited by 32 publications

References 28 publications

The role of GlpR repressor in Pseudomonas putida KT2440 growth and PHA production from glycerol

The role of GlpR repressor in Pseudomonas putida KT2440 growth and PHA production from glycerol

Glucose Metabolism in Xanthomonas campestris

Alginic acid synthesis in Pseudomonas aeruginosa mutants defective in carbohydrate metabolism

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