Title
Generation of a catR deficient mutant of P. putida KT2440 that produces cis, cis-muconate from benzoate at high rate and yield
Author
van Duuren, J.B.J.H.
Wijte, D.
Leprince, A.
Karge, B.
Puchalka, J.
Wery, J.
Dos Santos, V.A.P.M.
Eggink, G.
Mars, A.E.
Publication year
2011
Abstract
Pseudomonas putida KT2440-JD1 was derived from P. putida KT2440 after N-methyl-N'-nitro-N-nitrosoguanidine (NTG)-mutagenesis and exposure to 3-fluorobenzoate (3-FB). The mutant was no longer able to grow using benzoate as a sole carbon source, but co-metabolized benzoate to cis, cis-muconate during growth on glucose, which accumulated in the growth medium. The specific production rate (qpm) was 0.18±0.03g cis, cis-muconate/(gDCWh) in continuous cultures, and increased to 1.4g cis, cis-muconate/(gDCWh) during wash-out cultivation. Transcriptome analysis showed that the cat operon was not induced in P. putida KT2440-JD1 in the presence of 5mM benzoate, due to a point mutation in the highly conserved DNA binding domain of the transcriptional regulator (catR) of the cat operon. The ben operon was highly expressed in the presence of benzoate in the mutant and its parental strain. This operon contains PP_3166 (catA2), which was shown to be a second catechol 1,2-dioxygenase besides catA. P. putida KT2440-JD1 is the first cis, cis-muconate-accumulating mutant that was characterized at the genetic level. The specific production rate achieved is at least eight times higher than those reported for other cis, cis-muconate-producing strains. © 2011 Elsevier B.V.
Subject
Life
CBRN - CBRN Protection
EELS - Earth, Environmental and Life Sciences
Biology
3-Fluorobenzoate
Benzoate
Catechol 1,2-dioxygenase
Cis, cis-muconate
NTG mutagenesis
P. putida KT2440
3-Fluorobenzoate
Benzoate
Catechol 1 ,2-dioxygenase
Cis, cis-muconate
NTG mutagenesis
P. putida KT2440
Glucose
Mutagenesis
Phenols
Benzoic acid
Transcriptome
Article
DNA binding
Nonhuman
Operon
Priority journal
Pseudomonas putida
Transcription regulation
Pseudomonas putida
Benzoic acid, 532-32-1, 582-25-2, 65-85-0, 766-76-7
Carbon, 7440-44-0
Catechol 1,2 dioxygenase, 9027-16-1
Muconic acid, 505-70-4
To reference this document use:
http://resolver.tudelft.nl/uuid:9936501c-32df-4058-93d6-1fe9b7c663fc
DOI
https://doi.org/10.1016/j.jbiotec.2011.08.030
TNO identifier
442925
ISSN
0168-1656
Source
Journal of Biotechnology, 156 (3), 163-172
Document type
article