Print Email Facebook Twitter C1 compounds as auxiliary substrate for engineered Pseudomonas putida S12 Title C1 compounds as auxiliary substrate for engineered Pseudomonas putida S12 Author Koopman, F.W. de Winde, J.H. Ruijssenaars, H.J. TNO Kwaliteit van Leven Publication year 2009 Abstract The solvent-tolerant bacterium Pseudomonas putida S12 was engineered to efficiently utilize the C1 compounds methanol and formaldehyde as auxiliary substrate. The hps and phi genes of Bacillus brevis, encoding two key steps of the ribulose monophosphate (RuMP) pathway, were introduced to construct a pathway for the metabolism of the toxic methanol oxidation intermediate formaldehyde. This approach resulted in a remarkably increased biomass yield on the primary substrate glucose when cultured in C-limited chemostats fed with a mixture of glucose and formaldehyde. With increasing relative formaldehyde feed concentrations, the biomass yield increased from 35% (C-mol biomass/C-mol glucose) without formaldehyde to 91% at 60% relative formaldehyde concentration. The RuMP-pathway expressing strain was also capable of growing to higher relative formaldehyde concentrations than the control strain. The presence of an endogenous methanol oxidizing enzyme activity in P. putida S12 allowed the replacement of formaldehyde with the less toxic methanol, resulting in an 84% (C-mol/C-mol) biomass yield. Thus, by introducing two enzymes of the RuMP pathway, co-utilization of the cheap and renewable substrate methanol was achieved, making an important contribution to the efficient use of P. putida S12 as a bioconversion platform host. Subject Biology ResearchFood technologyAuxiliary substrateC1 compoundsPseudomonas putidaAuxiliary substrateBacillus brevisBiomass yieldC compoundsCo-utilizationControl strainFeed concentrationFormaldehyde concentrationsMethanol OxidationMonophosphatePseudomonas putidaRibuloseBacteriologyBiomassEnzyme activityEnzymesGlucoseMethanolSubstratesToxic materialsFormaldehydeformaldehydeglucosemethanolbacteriumbioengineeringbiomasschemostatenzyme activityformaldehydegene expressionmethanoloxidationsubstratearticlebacterial metabolismbiomass productionbiotransformationBrevibacillus breviscontrolled studyenzyme activityenzyme specificitygenetic engineeringmicrobial biomassnonhumanoxidationPseudomonas putidaBacillusBacterial ProteinsBiomassFormaldehydeGlucoseMetabolic Networks and PathwaysMethanolPseudomonas putidaBacteria (microorganisms)Brevibacillus brevisPseudomonas putida To reference this document use: http://resolver.tudelft.nl/uuid:a22ee989-ac84-4d15-baad-053d7500f80b DOI https://doi.org/10.1007/s00253-009-1922-y TNO identifier 93668 Source Applied microbiology and biotechnology, 83 (4), 705-713 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.