Abstract
In this work the biotransformation of α-pinene oxide to isonovalal using resting cells of Pseudomonas fluorescens NCIMB 11671 was evaluated in a membrane bioreactor for biotransformations (MBB). Since the membrane area required to obtain optimum productivities was calculated to be very large (1,000 m2 m−3), and not possible to fit into the laboratory reactor used, we initially evaluated performance with lower membrane areas (71 m2 m−3) in a batch system with both the substrate and product in the organic phase. This resulted in low productivities due to mass transfer limitations, so an optimum feeding rate of 0.1 g α-pinene oxide h−1 gcells −1 added directly to the reactor contents was determined in batch culture to minimise inhibition. The MBB was then operated continuously for the production of isonovalal, and a final concentration of 108 g l−1 was obtained in the organic reservoir after nearly 400 h of operation (0.32 g-isonovalal l−1 h−1), and the reaction was found not to be mass transfer limited. Finally, the relative viability of the cells was measured using fluorescent probes, and their half-life was found to be almost 2 months, confirming the ability of the MBB to facilitate biotransformations with inhibitory substrates and products.
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Amanullah A, Hewitt CJ, Nienow AW, Lee C, Chartrain BC, Buckland SW, Woodley JM (2003) Measurement of strain-dependent toxicity in the Indene bioconversion using multiparameter flow cytometry. Biotechnol Bioeng 81:405–420
Berger RG (1995) Aroma biotechnology. Springer, UK
Best DJ, Floyd NC, Magalhaes A, Burfield A, Rhodes PM (1987) Initial enzymatic steps in the degradation of α-pinene by Pseudomonas fluorescens NCIMB 11671. Biotechnol Bioeng 63:249–252
Boam A (1996) Novel bioreactor design for the biotransformation of hydrophobic compounds. Ph.D., University of London
Colocousi A (1994) Purification and sequencing of a-pinene monooxygenase from Pseudomonas fluorescens NCIMB 11671. Ph.D. University of London
Doig SD, Boam AT, Leak DJ, Livingston AG, Stuckey DC (1998) A membrane bioreactor for biotransformation of hydrophobic molecules. Biotechnol Bioeng 58:587–594
Doig SD, Boam AT, Livingston AG, Stuckey DC (1999) Mass transfer of hydrophobic solutes in solvent swollen silicone rubber membranes. J Membr Sci 154:127–140
Fichan I, Larroche C, Gros JB (1999) Water solubility, vapor pressure and activity coefficient of terpenes and terpenoids. J Chem Eng Data 44:56–62
Fontanille P, Le Fleche A, Larroche C (2002) Pseudomonas rhodisiae PF1: a new and efficient biocatalyst for production of isonovalal from α-pinene oxide. Biocatal Biotransform 20:413–421
Krings U, Berger RG (1998) Biotechnological production of flavours and fragrances. Appl Microbiol Biotechnol 49:1–8
Leon R, Fernandes P, Pinheiro HM, Cabral JMS (1998) Whole-cell biocatalysis in organic media. Enzyme Microb Technol 23:483–500
Lye GJ, Woodley JM (1999) Application of in situ product-removal techniques to biocatalytic process. Trends Biotechnol 17:395–402
Mattiasson B, Holst O (1991) Extractive bioconversions. Marcel Dekker, New York
Van der Werf MJ, de Bont JAM, Leak DJ (1997) Opportunities in microbial biotransformation of monoterpenes. Adv Biochem Eng/Biotechnol 55:147–177
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Boontawan, A., Stuckey, D.C. A membrane bioreactor for the biotransformation of α-pinene oxide to isonovalal by Pseudomonas fluorescens NCIMB 11671. Appl Microbiol Biotechnol 69, 643–649 (2006). https://doi.org/10.1007/s00253-005-0025-7
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DOI: https://doi.org/10.1007/s00253-005-0025-7