Print Email Facebook Twitter Generation of a catR deficient mutant of P. putida KT2440 that produces cis, cis-muconate from benzoate at high rate and yield 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 LifeCBRN - CBRN ProtectionEELS - Earth, Environmental and Life SciencesBiology3-FluorobenzoateBenzoateCatechol 1,2-dioxygenaseCis, cis-muconateNTG mutagenesisP. putida KT24403-FluorobenzoateBenzoateCatechol 1 ,2-dioxygenaseCis, cis-muconateNTG mutagenesisP. putida KT2440GlucoseMutagenesisPhenolsBenzoic acidTranscriptomeArticleDNA bindingNonhumanOperonPriority journalPseudomonas putidaTranscription regulationPseudomonas putidaBenzoic acid, 532-32-1, 582-25-2, 65-85-0, 766-76-7Carbon, 7440-44-0Catechol 1,2 dioxygenase, 9027-16-1Muconic 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 Files To receive the publication files, please send an e-mail request to TNO Library.