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Title (Primary) Constitutively solvent-tolerant Pseudomonas taiwanensis VLB120ΔCΔttgV supports particularly high styrene epoxidation activities when grown under glucose excess conditions
Author Volmer, J.; Lindmeyer, M.; Seipp, J.; Schmid, A.; Bühler, B.;
Journal Biotechnology and Bioengineering
Year 2019
Department SOMA;
Volume 116
Issue 5
Language englisch;
POF III (all) T15;
Keywords Biocatalytic oxyfunctionalization; biochemical engineering; energy metabolism; Pseudomonas; solvent tolerance; styrene epoxidation
UFZ wide themes MIKAT;
Abstract Solvent‐tolerant bacteria represent an interesting option to deal with substrate and product toxicity in bioprocesses. Recently, constitutive solvent tolerance was achieved for Pseudomonas taiwanensis VLB120 via knockout of the regulator TtgV, making tedious adaptation unnecessary. Remarkably, ttgV knockout increased styrene epoxidation activities of P. taiwanensis VLB120ΔC. With the aim to characterize and exploit the biocatalytic potential of P. taiwanensis VLB120ΔC and VLB120ΔCΔttgV, we investigated and correlated growth physiology, native styrene monooxygenase (StyAB) gene expression, whole‐cell bioconversion kinetics, and epoxidation performance. Substrate inhibition kinetics was identified, but was attenuated in two‐liquid phase bioreactor setups. StyA‐eGFP fusion enabled precise enzyme level monitoring without affecting epoxidation activity. Glucose limitation compromised styAB expression and specific activities (30‐40 U gCDW ‐1 for both strains), whereas unlimited batch cultivation enabled specific activities up to 180 U gCDW ‐1 for VLB120ΔCΔttgV strains, which is unrivaled for bioreactor‐based whole‐cell oxygenase biocatalysis. These extraordinarily high specific activities of constitutively solvent‐tolerant P. taiwanensis VLB120∆CttgV could be attributed to its high metabolic capacity, which also enabled high expression levels. This together with the high product yields on glucose and biomass obtained qualifies the VLB120∆ttgV strain as a highly attractive tool for the development of eco‐efficient oxyfunctionalization processes and redox biocatalysis in general.
ID 21489
Persistent UFZ Identifier https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=21489
Volmer, J., Lindmeyer, M., Seipp, J., Schmid, A., Bühler, B. (2019):
Constitutively solvent-tolerant Pseudomonas taiwanensis VLB120ΔCΔttgV supports particularly high styrene epoxidation activities when grown under glucose excess conditions
Biotechnol. Bioeng. 116 (5), 1089 - 1101