Modulation of Glucose Transport Causes Preferential Utilization of Aromatic Compounds in
            <i>Pseudomonas putida</i>
            CSV86

Basu, Ashoke Kumar; Shrivastava, Rahul; Basu, Bhakti; Apte, Shree Kumar; Phale, Prashant S.

doi:10.1128/jb.01235-07

Cited by 32 publications

(40 citation statements)

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“…P. putida CSV86 preferentially utilizes aromatics or organic acids over glucose and co-metabolizes aromatics and organic acids . Previously, we have demonstrated that glucose induces glucose-6-phosphate dehydrogenase (Zwf), an intracellular glucose-metabolizing enzyme ) and a 43 kDa glucosebinding periplasmic space protein (Basu et al, 2007). Both Zwf and the 43 kDa protein are repressed by aromatics as well as by organic acids Basu et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

“…Outer-membrane proteins were resolved by SDS-PAGE (10 %) as described by Laemmli (1970). Outer-membrane protein fractions were also subjected to 2D gel electrophoresis (Basu et al, 2007;Joshi et al, 2004;O'Farrell, 1975) followed by LC-MS/MS analysis (ESI-TRAP, trypsin digest; peptide mass tolerance±2 Da; fragment mass tolerance, 0.8 Da) as described by Basu et al (2007). The data were analysed by using the Mascot protein identification system (Matrix Science) with the database from The Institute of Genomic Research (TIGR; http://www.jcvi.org/).…”

Section: Methodsmentioning

confidence: 99%

“…In CSV86, glucose is taken up by active transport and is metabolized via an inducible intracellular phosphorylative pathway and not by an oxidative pathway . A 43 kDa periplasmic glucose-binding protein has earlier been reported to be induced during growth on glucose and suppressed during growth on aromatics and organic acids (Basu et al, 2007). The present study investigates the outermost transport component and its possible involvement in such regulation.…”

Section: Introductionmentioning

confidence: 99%

“…However, toluene did not affect the oxidative route of glucose metabolism (del Castillo & Ramos, 2007). The phosphorylative mode of glucose metabolism requires its transport inside the cell and involves: (i) an inducible outer-membrane protein (Hancock & Carey, 1980;Saravolac et al, 1991;Wylie & Worobec, 1995), (ii) an inducible periplasmic glucose-binding protein (Basu et al, 2007;Sly et al, 1993;Stinson et al, 1977) and (iii) a putative inner-membrane protein(s) (Adewoye & Worobec, 2000). So far, to our knowledge, all three components have not been reported from a single Pseudomonas strain.…”

Section: Introductionmentioning

confidence: 99%

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Repression of the glucose-inducible outer-membrane protein OprB during utilization of aromatic compounds and organic acids in Pseudomonas putida CSV86

et al. 2011

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Pseudomonas putida CSV86 shows preferential utilization of aromatic compounds over glucose. Protein analysis and [ 14 C]glucose-binding studies of the outer membrane fraction of cells grown on different carbon sources revealed a 40 kDa protein that was transcriptionally induced by glucose and repressed by aromatics and succinate. Based on 2D gel electrophoresis and liquid chromatography-tandem mass spectrometry analysis, the 40 kDa protein closely resembled the porin B of P. putida KT2440 and carbohydrate-selective porin OprB of various Pseudomonas strains. The purified native protein (i) was estimated to be a homotrimer of 125 kDa with a subunit molecular mass of 40 kDa, (ii) displayed heat modifiability of electrophoretic mobility, (iii) showed channel conductance of 166 pS in 1 M KCl, (iv) permeated various sugars (mono-, di-and trisaccharides), organic acids, amino acids and aromatic compounds, and (v) harboured a glucosespecific and saturable binding site with a dissociation constant of 1.3 mM. These results identify the glucose-inducible outer-membrane protein of P. putida CSV86 as a carbohydrate-selective protein OprB. Besides modulation of intracellular glucose-metabolizing enzymes and specific glucose-binding periplasmic space protein, the repression of OprB by aromatics and organic acids, even in the presence of glucose, also contributes significantly to the strain's ability to utilize aromatics and organic acids over glucose.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Repression of the glucose-inducible outer-membrane protein OprB during utilization of aromatic compounds and organic acids in Pseudomonas putida CSV86

et al. 2011

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“…In E. coli, the cAMP receptor protein Crp is involved in CCR of PAA degradation (Ferrández et al, 2000;Kimata et al, 1997;Notley-McRobb et al, 1997). However, the level of cAMP in P. putida remains constant regardless of the carbon source and cAMP is unlikely to be involved in CCR in this species (Basu et al, 2007;Siegel et al, 1977). CCR of PAA metabolism appears to be unaffected by organic acids, such as pyruvate or succinate (Kim et al, 2007), suggesting that CCR is generated by glucose intermediates (Fig.…”

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confidence: 99%

Intracellular 2-keto-3-deoxy-6-phosphogluconate is the signal for carbon catabolite repression of phenylacetic acid metabolism in Pseudomonas putida KT2440

et al. 2009

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The growth pattern of Pseudomonas putida KT2440 in the presence of glucose and phenylacetic acid (PAA), where the sugar is used in preference to the aromatic compound, suggests that there is carbon catabolite repression (CCR) of PAA metabolism by glucose or gluconate. Furthermore, CCR is regulated at the transcriptional level. However, this CCR phenomenon does not occur in PAA-amended minimal medium containing fructose, pyruvate or succinate. We previously identified 2-keto-3-deoxy-6-phosphogluconate (KDPG) as an inducer of glucose metabolism, and this has led to this investigation into the role of KDPG as a signal compound for CCR. Two mutant strains, the edd mutant (non-KDPG producer) and the eda mutant (KDPG overproducer), grew in the presence of PAA but not in the presence of glucose. The edd mutant utilized PAA even in the presence of glucose, indicating that CCR had been abolished. This observation has additional support from the finding that there is high phenylacetyl-CoA ligase activity in the edd mutant, even in the presence of glucose+PAA, but not in wild-type cells under the same conditions. Unlike the edd mutant, the eda mutant did not grow in the presence of glucose+PAA. Interestingly, there was no uptake and/or metabolism of PAA in the eda mutant cells under the same conditions. Targeted disruption of PaaX, a repressor of the PAA operon, had no effect on CCR of PAA metabolism in the presence of glucose, suggesting that there is another transcriptional repression system associated with the KDPG signal. This is the first study to demonstrate that KDPG is the true CCR signal of PAA metabolism in P. putida KT2440. INTRODUCTIONIn Pseudomonas putida, the phenylacetic acid (PAA) catabolic gene clusters consist of 15 genes that are organized into five contiguous operons (del Peso-Santos et al., 2008; Di Gennaro et al., 2007; Jiménez et al., 2002;Luengo et al., 2001;Olivera et al., 1998). The PAA pathway ( Fig. 1) is a major route of a complex functional unit that catalyses the transformation of structurally related compounds such as styrene, tropic acid, ethylbenzene and 2-phenylethylalanine into phenylacetyl-CoA (PA-CoA), a common intermediate (Luengo et al., 2001). It has been reported that PA-CoA, and not PAA, is the true inducer of the PAA catabolic pathway and that the PaaX repressor controls PAA-catabolic gene expression (Ferrández et al., 2000;García et al., 2000). Binding of PaaX to its cognate DNA binding sites is abolished when the inducer, PA-CoA, interacts with the PaaX protein (Bartolomé-Martín et al., 2004; del Peso-Santos et al., 2006; Ferrández et al., 2000;García et al., 2000). Interestingly, along with the StyR/StyS system, PaaX also regulates the styrene degradation pathway by binding to the promoter region of the styrene upper pathway in Pseudomonas sp. strain Y2 (del PesoSantos et al., 2006). PaaX is also involved in the transcriptional regulation of the pac gene, which encodes penicillin G acylase in Escherichia coli W (Galán et al., 2004;Kim et al., 2004). PaaX is involved in the co...

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Production of p‐hydroxybenzoic acid from p‐coumaric acid by Burkholderia glumae BGR1

Jung

Kim

Jung

et al. 2015

Biotech & Bioengineering

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p-Coumaric acid (pCA) is abundant in biomass with low lignin content, such as straw and stubble from rye, wheat, and barley. pCA can be isolated from biomass and used for the synthesis of aromatic hydrocarbons. Here, we report engineering of the natural pathway for conversion of pCA into p-hydroxybenzoic acid (pHBA) to increase the amount of pHBA that accumulates more than 100-fold. Burkholderia glumae strain BGR1 (BGR1) grows efficiently on pCA as a sole carbon source via a CoA-dependent non-β-oxidation pathway. This pathway removes two carbons from pCA as acetyl-CoA yielding p-hydroxybenzaldehyde and subsequently oxidizes it to pHBA. To increase the amount of accumulated pHBA in BGR1, we first deleted two genes encoding enzymes that degrade pHBA in the β-ketoadipate pathway. At 10 mM of pCA, the double deletion mutant BGR1_PB4 (Δphb3hΔbcl) accumulated pHBA with 95% conversion, while the control BGR1 accumulated only with 11.2% conversion. When a packed bed reactor containing immobilized BGR1_PB4 cells was operated at a dilution rate 0.2 h(-1) , the productivity of pHBA was achieved at 9.27 mg/L/h for 134 h. However, in a batch reactor at 20 mM pCA, growth of BGR1_PB4 was strongly inhibited, resulting in a low conversion of 19.3%. To further increase the amount of accumulated pCA, we identified the first enzyme in the pathway, p-hydroxcinnmaoyl-CoA synthetase II (phcs II), as the rate-limiting enzyme. Over expression of phcs II using a Palk promoter in a batch reaction at 20 mM of pCA yielded 99.0% conversion to pHBA, which is the highest concentration of pHBA ever reported using a biological process. Biotechnol. Bioeng. 2016;113: 1493-1503. © 2015 Wiley Periodicals, Inc.

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Modulation of Glucose Transport Causes Preferential Utilization of Aromatic Compounds in Pseudomonas putida CSV86

Cited by 32 publications

References 47 publications

Repression of the glucose-inducible outer-membrane protein OprB during utilization of aromatic compounds and organic acids in Pseudomonas putida CSV86

Repression of the glucose-inducible outer-membrane protein OprB during utilization of aromatic compounds and organic acids in Pseudomonas putida CSV86

Intracellular 2-keto-3-deoxy-6-phosphogluconate is the signal for carbon catabolite repression of phenylacetic acid metabolism in Pseudomonas putida KT2440

Production of p‐hydroxybenzoic acid from p‐coumaric acid by Burkholderia glumae BGR1

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