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Generation of a catR deficient mutant of P. putida KT2440 that produces cis, cis-muconate from benzoate at high rate and yield

Paper ID Volume ID Publish Year Pages File Format Full-Text
23732 43468 2011 10 PDF Available
Title
Generation of a catR deficient mutant of P. putida KT2440 that produces cis, cis-muconate from benzoate at high rate and yield
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 ± 0.03 g cis, cis-muconate/(gDCW h) in continuous cultures, and increased to 1.4 g cis, cis-muconate/(gDCW h) during wash-out cultivation. Transcriptome analysis showed that the cat operon was not induced in P. putida KT2440-JD1 in the presence of 5 mM 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.

► Pseudomonas putida KT2440-JD1 produces cis, cis-muconate with high specific productivity. ► P. putida KT2440-JD1 is the first genetically characterized muconate producer. ► The gene PP_3166 (catA2) of the ben operon encodes a second catechol 1,2-dioxygenase.

Keywords
Benzoate; Catechol 1,2-dioxygenase; cis, cis-muconate; 3-Fluorobenzoate; NTG mutagenesis; P. putida KT2440
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Generation of a catR deficient mutant of P. putida KT2440 that produces cis, cis-muconate from benzoate at high rate and yield
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 156, Issue 3, 10 December 2011, Pages 163–172
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us