Effect of temperature on diauxic growth with glucose and organic acids in Pseudomonas fluorescens

Lynch, William H.; Franklin, M.

doi:10.1007/bf00415721

Cited by 37 publications

(22 citation statements)

References 26 publications

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“…Whatever its mechanism, it is likely that the same catabolite repression control system exists in the fluorescent pseudomonads, since we were able to identify proteins which were antigenically similar to Crc in P. putida and P. fluorescens but not in E. coli. The existence of a common mechanism of catabolite repression control in these closely related species is not surprising, since repression by organic acids has been observed in all three species (16,42).…”

Section: Discussionmentioning

confidence: 93%

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The nucleotide sequence of the Pseudomonas aeruginosa pyrE-crc-rph region and the purification of the crc gene product

et al. 1996

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The gene (crc) responsible for catabolite repression control in Pseudomonas aeruginosa has been cloned and sequenced. Flanking the crc gene are genes encoding orotate phosphoribosyl transferase (pyrE) and RNase PH (rph). New crc mutants were constructed by disruption of the wild-type crc gene. The crc gene encodes an open reading frame of 259 amino acids with homology to the apurinic/apyrimidinic endonuclease family of DNA repair enzymes. However, crc mutants do not have a DNA repair phenotype, nor can the crc gene complement Escherichia coli DNA repair-deficient strains. The crc gene product was overexpressed in both P. aeruginosa and in E. coli, and the Crc protein was purified from both. The purified Crc proteins show neither apurinic/ apyrimidinic endonuclease nor exonuclease activity. Antibody to the purified Crc protein reacted with proteins of similar size in crude extracts from Pseudomonas putida and Pseudomonas fluorescens, suggesting a common mechanism of catabolite repression in these three species.The genus Pseudomonas is noteworthy for its diversity in habitat and physiology. Some Pseudomonas strains are able to use over 100 organic compounds as the sole or principal source of carbon. Pseudomonas aeruginosa can utilize at least 80 different organic compounds (41). A mechanism of catabolite repression control exists in these organisms (18) which prevents them from wasting energy maintaining the enzymes for all these catabolic pathways and ensures the preferential utilization of the most efficient source of carbon and energy. Such a regulatory mechanism has also been identified in the enteric bacteria (20) and in Bacillus spp. (6).In the enteric bacteria, the molecular mechanism of catabolite repression control involves a catabolite activator protein (Cap) which, when bound to cyclic AMP (cAMP), interacts with promoter regions of regulated genes to facilitate the binding of RNA polymerase, thereby initiating transcription (20). In the presence of glucose, the cAMP pool is lowered, Cap is not bound, and the regulated genes are not transcribed (i.e., they are repressed). The effect of glucose on cAMP pools is mediated by components of the phosphoenolpyruvate phosphotransferase system which also serve to activate adenylate cyclase (33). Since the initial identification of these regulatory components, catabolite repression has proven to be a global mechanism in the enteric bacteria, affecting at least 28 separate promoters which regulate biosynthetic as well as catabolic operons (7). It also has been found to act as a negative regulator as well as a positive regulator (1,7,23,24).In Bacillus spp. catabolite repression is not mediated by glucose, nor is cAMP involved (6,37,40). Genetic evidence has implicated the catabolite control protein (CcpA), a member of the GalR family of repressor proteins (6), Hpr, a component of the phosphoenolpyruvate phosphotransferase system (9), and a cis-acting DNA sequence (CRE) (15,48). Recent work has shown that HPr specifically phosphorylated at Ser-46 forms a complex wit...

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Section: Discussionmentioning

confidence: 93%

“…Bacterial strains and plasmids used in this study are listed in Table 1. P. aeruginosa was grown at 37ЊC in basal salts medium (BSM) (16) or in Luria-Bertani (LB) medium (22). Pseudomonas fluorescens and P. putida were grown at 30ЊC in LB medium.…”

Section: Methodsmentioning

confidence: 99%

The nucleotide sequence of the Pseudomonas aeruginosa pyrE-crc-rph region and the purification of the crc gene product

et al. 1996

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“…Previous studies with different bacterial species showed that occurrence of diauxic growth behavior critically depends on the history of the inoculum (5,9,11). Therefore, batch cultures were first inoculated with cells pregrown with acetate.…”

Section: Resultsmentioning

confidence: 99%

“…In such cases, enzymes involved in the metabolism of the less preferred substrate occur after exhaustion of the preferred substrate (1,4,5,7,9,11). However, if the inoculum is grown on the less favored substrate, the lag between the two phases of diauxic growth becomes obscured.…”

Section: Discussionmentioning

confidence: 99%

Control of diauxic growth of Azotobacter vinelandii on acetate and glucose

1990

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Batch cultures of Azotobacter vinelandii were inoculated with cells pregrown on either acetate or glucose. When they were subsequently grown on a mixture of acetate and glucose, typical diauxic growth was observed, with preferential uptake of acetate in the first and glucose in the second phase of growth. Extracts from acetate-pregrown cells exhibited high acetate kinase activity in the first phase of growth. This activity decreased and activities of the two glucose enzymes glucose 6-phosphate dehydrogenase and glyceraldehyde 3-phosphate dehydrogenase increased in the second phase. Extracts from glucose-pregrown cells exhibited high initial activities of the two glucose enzymes, which decreased while acetate kinase activity increased in the first phase of growth. Again, in the second phase, activities of the two glucose enzymes increased and acetate kinase activity decreased. In any case, isocitrate dehydrogenase activity varied only slightly and unspecifically. The differences in enzyme activity and the constancy of isocitrate dehydrogenase were confirmed by experiments with either acetate-or glucose-limited chemostats. In chemostats in which both of the substrates were limiting, all of the enzymes displayed significant activities. Glucose 6-phosphate dehydrogenase activity was inhibited by acetyl coenzyme A and acetyl phosphate but not by acetate. It is proposed that diauxic growth is based on the control of enzymes involved in acetate or glucose dissimilation by which acetate or its metabolites control the expression and activity of glucose enzymes.Azotobacter vinelandii is an aerobic dinitrogen (N2)-fixing organism. Oxidation of a variety of organic substrates provides energy and reducing power for biomass production, including reduction of N2 to ammonia and its assimilation into cell material. Expression of the N2-fixing enzyme system, the nitrogenase complex, is controlled by the availability of fixed N and an organic substrate serving as the energy and carbon source. Previously, we demonstrated that upon increase of the C/N ratio in the culture medium, cells of A. vinelandii increased the rates of substrate consumption and, consequently, respiration before the threshold C/N ratio for nitrogenase expression was reached (2, 3). As a result, A. vinelandii exhibits unusually high rates of substrate consumption and oxygen uptake when fixing N2. However, despite the high demand for organic substrates, the organisms perform diauxic growth when supplied with acetate and glucose (6).Diauxic growth can be observed when heterotrophic bacteria grow in batch culture on sufficiently high concentrations of more than one carbon and energy source (7). In most cases, the substrate which permits the highest growth rate is used up first. Thus, the more preferred and less preferred substrates are used up sequentially. As a result, diauxic growth is characterized by two exponential phases of growth separated by a phase of no net increase in biomass. Harder and Dijkhuizen (7) distinguished at least three levels at which utiliza...

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“…The duration of the lag is significantly reduced with an inoculum prepared on mannopine. In some (10,17,20), although not all (26) figure. lag phase was similarly decreased with an inoculum prepared on the less preferred substrate.…”

Section: Materuils and Methodsmentioning

confidence: 97%

Diauxic growth of Agrobacterium tumefaciens 15955 on succinate and mannopine

1992

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Agrobacterium tumefaciens induces crown gall tumors in a wide range of plants by transfer of DNA from its tumorinducing (Ti) plasmid into the chromosome of the host plant (1, 16). The transferred DNA contains genes for production of phytohormones which are responsible for rapid growth of the transformed plant cells as a tumor. The transferred DNA also contains genes whose expression in the plant brings about reductive conjugation of amino acids with carbonyl compounds of primary metabolism (27). The conjugated amino acids, called opines, are secondary metabolites no longer available for utilization by the plant. The nontransferred portion of the Ti plasmid contains genes permitting bacterial utilization of opines as sources of both carbon and nitrogen for growth. Ti plasmid genes required for opine utilization by A. tumefaciens are inducible by the appropriate opine. Ti plasmids are biochemically distinguishable by the family of opines whose synthesis they encode. Opine families differ in the nature of amino acid conjugated (basic amino acids, leucine, or asparagine), the identity of the carbonyl partner (glucose, pyruvic acid, or a-ketoglutaric acid), and the chirality of the new asymmetric center produced in the reductive conjugation (2, 4-6, 25). The mannityl family of opines is synthesized in the plant tumor by condensation of glucose and glutamine followed by the Amadori rearrangement to NT-l'-(l'-deoxy-D-fructosyl)-L-glutamine and subsequent reduction to N'-1'-(1'-deoxy-D-mannityl)-Lglutamine (mannopine) (11). The transferred DNA of some Ti plasmids also codes for a lactonizing enzyme which converts mannopine into agropine (7). Both mannopine and agropine are slowly converted nonenzymatically to the lactam agropinic acid. Mannopinic acid also occurs in tumors together with the three other mannityl opines. Catabolism of the four mannityl opines is encoded on the Ti plasmids (9). Some of the Ti plasmids which determine biosynthesis and catabolism of the mannityl opines also determine the metabolism of octopine, which is an imino diacid type of opine * Corresponding author.composed of arginine and pyruvate moieties (27). Yet other Ti plasmids code for the production and degradation of mannityl opines and S,S-leucinopine (2, 4). Various types of the related root-inducing (Ri) plasmid of Agrobactenum rhizogenes also determine mannopine biosynthesis by transformed plant cells and utilization by bacteria (23).Catabolism of mannopinic acid by A. tumefaciens 15955 involves cleavage of the molecule at the C-N bond between the amino acid and the sugar moieties, generating mannose and also probably glutamate (8, 9). Likewise, studies of mannityl analogs comprising a nonmetabolizable sugar moiety suggest cleavage at the bond between the amino acid and the sugar components of the molecule (3, 22). However, nothing is known about degradative pathways for the three other mannityl opines, including mannopine.In studies dealing with bacterial catabolism of mannopine and other opines, the test substrate is generally present ...

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Effect of temperature on diauxic growth with glucose and organic acids in Pseudomonas fluorescens

Cited by 37 publications

References 26 publications

The nucleotide sequence of the Pseudomonas aeruginosa pyrE-crc-rph region and the purification of the crc gene product

The nucleotide sequence of the Pseudomonas aeruginosa pyrE-crc-rph region and the purification of the crc gene product

Control of diauxic growth of Azotobacter vinelandii on acetate and glucose

Diauxic growth of Agrobacterium tumefaciens 15955 on succinate and mannopine

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