Prediction of the adaptability of Pseudomonas putida DOT-T1E to a second phase of a solvent for economically sound two-phase biotransformations.
The strain Pseudomonas putida DOT-T1E was tested for its ability to tolerate second phases of different alkanols for their use as solvents in two-liquid-phase biotransformations. Although 1-decanol showed an about 10-fold higher toxicity to the cells than 1-octanol, the cells were able to adapt completely to 1-decanol only and could not be adapted in order to grow stably in the presence of a second phase of 1-octanol. The main explanation for this observation can be seen in the higher water and membrane solubility of 1-octanol. The hydrophobicity (log P) of a substance correlates with a certain partitioning of that compound into the membrane. Combining the log P value with the water solubility, the maximum membrane concentration of a compound can be calculated. With this simple calculation, it is possible to predict the property of an organic chemical for its potential applicability as a solvent for two-liquid-phase biotransformations with solvent-tolerant P. putida strains. Only compounds that show a maximum membrane concentration of less than 400 mM, such as 1-decanol, seem to be tolerated by these bacterial strains when applied in supersaturating concentrations to the medium. Taking into consideration that a solvent for a two-liquid-phase system should possess partitioning properties for potential substrates and products of a fine chemical synthesis, it can be seen that 1-decanol is a suitable solvent for such biotransformation processes. This was also demonstrated in shake cultures, where increasing amounts of a second phase of 1-decanol led to bacteria tolerating higher concentrations of the model substrate 3-nitrotoluene. Transferring this example to a 5-liter-scale bioreactor with 10% (vol/vol) 1-decanol, the amount of 3-nitrotoluene tolerated by the cells is up to 200-fold higher than in pure aqueous medium. The system demonstrates the usefulness of two-phase biotransformations utilizing solvent-tolerant bacteria.
Neumann
Grit
G
Department of Bioremediation, Centre for Environmental Research Leipzig-Halle (UFZ), Permoserstr. 15, 04318 Leipzig, Germany.
Kabelitz
Nadja
N
Zehnsdorf
Andreas
A
Miltner
Anja
A
Lippold
Holger
H
Meyer
Daniel
D
Schmid
Andreas
A
Heipieper
Hermann J
HJ
D020003:1-Octanol / Q000378:metabolism*; D000222:Adaptation, Physiological; D019149:Bioreactors; D001709:Biotechnology / Q000379:methods; D003470:Culture Media; D005233:Fatty Alcohols / Q000378:metabolism*; D005285:Fermentation; D011237:Predictive Value of Tests; D016958:Pseudomonas putida / Q000254:growth & development* / Q000378:metabolism / Q000502:physiology; D012997:Solvents / Q000378:metabolism*; D014050:Toluene / Q000031:analogs & derivatives* / Q000378:metabolism; D014867:Water
D003470:Culture Media; D005233:Fatty Alcohols; D012997:Solvents; D014867:Water; C029956:3-nitrotoluene; D014050:Toluene; C029383:n-decyl alcohol; D020003:1-Octanol