Solvent-tolerant microorganisms are useful in biotransformations with whole cells in two-phase solvent-water systems. The results presented here describe the effects that organic solvents have on the growth of these organisms. The maximal growth rate of Pseudomonas putida S12, 0.8 h−1, was not affected by toluene in batch cultures, but in chemostat cultures the solvent decreased the maximal growth rate by nearly 50%. Toluene, ethylbenzene, propylbenzene, xylene, hexane, and cyclohexane reduced the biomass yield, and this effect depended on the concentration of the solvent in the bacterial membrane and not on its chemical structure. The dose response to solvents in terms of yield was linear up to an approximately 200 mM concentration of solvent in the bacterial membrane, both in the wild type and in a mutant lacking an active efflux system for toluene. Above this critical concentration the yield of the wild type remained constant at 0.2 g of protein/g of glucose with increasing concentrations of toluene. The reduction of the yield in the presence of solvents is due to a maintenance higher by a factor of three or four as well as to a decrease of the maximum growth yield by 33%. Therefore, energy-consuming adaptation processes as well as the uncoupling effect of the solvents reduce the yield of the tolerant cells.

https://aem.asm.org/content/aem/65/6/2631.full.pdf

Effect of Organic Solvents on the Yield of Solvent-Tolerant Pseudomonas putida S12

Application of biotransformations to the synthesis of industrial chemicals is in part limited by a number of process challenges. We discuss the conversion of toxic, poorly water-soluble organic substrates by whole cells, using as an illustrative example the specific hydroxylation of toluene to toluenecis-glycol byPseudomonas putida UV4. Toxic effects may be eliminated through the introduction of tetradecane, to partition toluene away from the biocatalyst, to give product concentrations of 30–60 gL−1, in a two-liquid-phase reactor. The operational limits of this system have been experimentally determined and are presented in the form of windows of operation.

Determination of reactor operation for the microbial hydroxylation of toluene in a two-liquid phase process

Expression and longevity of toluene dioxygenase in Pseudomonas putida F1 induced at different dissolved oxygen concentrations.

Design of a control system for biotransformation of toxic substrates: toluene hydroxylation by Pseudomonas putida UV4

The use of oxygen uptake rate measurements to control the supply of toxic substrate: toluene hydroxylation by Pseudomonas putida UV4.

A constitutive blocked mutant (UV4) of Pseudomonas putida was grown in a 2.5-l fermentor on a mineral salts medium. Glucose was fed normally over an 18-h period. Gluconate reached about 5 gl−1 in the medium and then fell to zero as it was utilised. The maximum toluene dioxygenase specific activity (2 g g−1 h−1) was obtained over the last 6 h of the fermentation when the pH was fully controlled. In fermentations done at low dissolved O2 tension (DOT) values there was an overall reduction in the cellular enzyme level. When stored at 4°C in phosphate buffer, pH 7.0, harvested bacteria lost half their activity in about 90 h.

The production of Pseudomonas putida for the hydroxylation of toluene to its cis-glycol

Evaluation of process options for the biotransformation of toluene to toluene cis-glycol by Pseudomonas putida

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The production of Pseudomonas putida for the hydroxylation of toluene to its cis-glycol

Evaluation of process options for the biotransformation of toluene to toluene cis-glycol by Pseudomonas putida