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 Research
Food technology
Auxiliary substrate
C1 compounds
Pseudomonas putida
Auxiliary substrate
Bacillus brevis
Biomass yield
C compounds
Co-utilization
Control strain
Feed concentration
Formaldehyde concentrations
Methanol Oxidation
Monophosphate
Pseudomonas putida
Ribulose
Bacteriology
Biomass
Enzyme activity
Enzymes
Glucose
Methanol
Substrates
Toxic materials
Formaldehyde
formaldehyde
glucose
methanol
bacterium
bioengineering
biomass
chemostat
enzyme activity
formaldehyde
gene expression
methanol
oxidation
substrate
article
bacterial metabolism
biomass production
biotransformation
Brevibacillus brevis
controlled study
enzyme activity
enzyme specificity
genetic engineering
microbial biomass
nonhuman
oxidation
Pseudomonas putida
Bacillus
Bacterial Proteins
Biomass
Formaldehyde
Glucose
Metabolic Networks and Pathways
Methanol
Pseudomonas putida
Bacteria (microorganisms)
Brevibacillus brevis
Pseudomonas 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