Rhodospirillum rubrum: utilization of condensed corn solubles for poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) production.

AIMS: This study sought to develop a less expensive medium for growth of the polyhydroxyalkanoate-producing bacterium Rhodospirillum rubrum from the ethanol production coproduct, condensed corn solubles (CCS). METHODS AND RESULTS: Small-scale trials using R. rubrum were performed in aerated or anaerobic stoppered serum bottles filled with media. The CCS (240 g l(-1)) achieved a maximum cell density and growth rate comparable with the defined supplemented malate-ammonium medium (mSMN) or tryptic soy broth. Microaerophilic solubles medium cultures exhibited significantly higher maximum cell densities and growth rates than did strictly anaerobic cultures; while illumination, nickel or biotin addition had no effect. Growth of R. rubrum in a pH controlled bioreactor was significantly better in CCS (240 g l(-1)) than in mSMN medium and supported production of 0.36% (cell dry weight) poly-(3-hydroxybutyrate-Co-3-hydroxyvalerate) after 24 h. CONCLUSIONS: A CCS medium was devised that supported R. rubrum growth for biopolymer production as effective as the defined medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that a more economical medium can be developed for biopolymer production using a low value coproduct from ethanol production. The impact is that this inexpensive solubles medium may make it more economical to produce the biopolymer on a commercial scale.

Control of pyrimidine synthesis in Pseudomonas fragi.

AIMS: To study the regulation of the de novo pyrimidine biosynthetic enzymes in the food-spoilage agent Pseudomonas fragi ATCC 4973. METHODS AND RESULTS: The de novo pyrimidine biosynthetic enzymes were measured in extracts of Ps. fragi ATCC 4973 cells and of cells from auxotrophs deficient for dihydroorotase or OMP decarboxylase activity. Pyrimidine biosynthetic enzyme activities in ATCC 4973 were influenced by pyrimidine supplementation to the culture medium. The pyrimidine limitation of each auxotroph elevated the de novo enzyme activities, indicating that this pathway may be repressible by a pyrimidine-related compound. Aspartate transcarbamoylase activity in ATCC 4973 was inhibited in vitro by pyrophosphate and purine or pyrimidine nucleotides. CONCLUSIONS: Pyrimidine synthesis in Ps. fragi appeared to be controlled at the transcriptional level and at the level of activity for aspartate transcarbamoylase. Its transcriptional regulation seemed to be more highly controlled than what was observed in the closely related species Pseudomonas putida and Pseudomonas fluorescens. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that pyrimidine synthesis is regulated in Ps. fragi. This could prove useful to future studies examining its biological control and its taxonomic assignment.

Pyrimidine biosynthesis in Pseudomonas oleovorans.

AIMS: To investigate the regulation of de novo pyrimidine biosynthesis in the polyhydroxyalkanoate-producing bacterium Pseudomonas oleovorans at the level of enzyme synthesis and at the level of aspartate transcarbamoylase activity. METHODS AND RESULTS: The effect of pyrimidine supplementation on the pyrimidine biosynthetic pathway enzyme activities was analysed relative to carbon source. Two uracil auxotrophs of P. oleovorans were isolated that were deficient for aspartate transcarbamoylase or dihydroorotase activity. Pyrimidine limitation of these auxotrophs increased the de novo pathway activities to varying degrees depending on the pathway mutation and the carbon source utilized. At the level of aspartate transcarbamoylase activity, pyrophosphate and uridine ribonucleotides were found to be strongly inhibitory of the Ps. oleovorans enzyme. CONCLUSIONS: Pyrimidine biosynthesis is regulated in Ps. oleovorans. Taxonomically, the regulation of the pyrimidine biosynthetic pathway appeared dissimilar from previously studied Pseudomonas species. SIGNIFICANCE AND IMPACT OF THE STUDY: New insights regarding the regulation of nucleic acid metabolism are provided that could prove significant during the genetic manipulation of Ps. oleovorans to increase the synthesis of polyhydroxyalkanoates.

Ability of casamino acids to support gellan production by Sphingomonas paucimobilis ATCC 31461.

The ability of casamino acids and vitamin-assay casamino acids to support gellan production by Sphingomonas paucimobilis ATCC 31461 was examined in a medium containing glucose or corn syrup as the carbon source relative to yeast extract supplementation. When glucose or corn syrup served as the carbon source, the presence of yeast extract in the growth medium stimulated gellan production by strain ATCC 31461 on casamino acids. Using vitamin-assay casamino acids as the nitrogen source, the addition of vitamins lowered gellan synthesis by glucose-grown cells regardless of yeast extract supplementation while gellan elaboration by corn syrup-grown strain ATCC 31461 cells could only be increased by supplementing vitamins into medium lacking yeast extract. Independent of carbon source, the absence of yeast extract in the medium reduced biomass production. Biomass production by the strain grown on either carbon source was increased by supplementing vitamins in the medium containing yeast extract.