Statistical Optimisation of Rhamnolipid Production using a Pseudomonas putida Strain Cultivated on Renewable Carbon Sources of Waste Vegetable Oils

2020 ◽  
Vol 57 (1) ◽  
pp. 13-21 ◽  
Author(s):  
Zulfiqar Ali Raza ◽  
Zafar M. Khalid ◽  
Naseer Ahmad ◽  
Bushra Tehseen
Keyword(s):  
Pseudomonas Putida ◽  
Vegetable Oils ◽  
Carbon Sources ◽  
Rhamnolipid Production

scholarly journals Burkholderia thailandensis E264 as a promising safe rhamnolipids’ producer towards a sustainable valorization of grape marcs and olive mill pomace

2021 ◽  
Author(s):  
Alif Chebbi ◽  
Massimiliano Tazzari ◽  
Cristiana Rizzi ◽  
Franco Hernan Gomez Tovar ◽  
Sara Villa ◽  
...  
Keyword(s):  
Energy Source ◽  
Olive Oil ◽  
Low Cost ◽  
Carbon Sources ◽  
Burkholderia Thailandensis ◽  
Rhamnolipid Production ◽  
Key Points ◽  
Wide Range ◽  
Long Chain ◽  
Olive Mill

Abstract Within the circular economy framework, our study aims to assess the rhamnolipid production from winery and olive oil residues as low-cost carbon sources by nonpathogenic strains. After evaluating various agricultural residues from those two sectors, Burkholderia thailandensis E264 was found to use the raw soluble fraction of nonfermented (white) grape marcs (NF), as the sole carbon and energy source, and simultaneously, reducing the surface tension to around 35 mN/m. Interestingly, this strain showed a rhamnolipid production up to 1070 mg/L (13.37 mg/g of NF), with a higher purity, on those grape marcs, predominately Rha-Rha C14-C14, in MSM medium. On olive oil residues, the rhamnolipid yield of using olive mill pomace (OMP) at 2% (w/v) was around 300 mg/L (15 mg/g of OMP) with a similar CMC of 500 mg/L. To the best of our knowledge, our study indicated for the first time that a nonpathogenic bacterium is able to produce long-chain rhamnolipids in MSM medium supplemented with winery residues, as sole carbon and energy source. Key points • Winery and olive oil residues are used for producing long-chain rhamnolipids (RLs). • Both higher RL yields and purity were obtained on nonfermented grape marcs as substrates. • Long-chain RLs revealed stabilities over a wide range of pH, temperatures, and salinities

scholarly journals Simultaneous Catabolite Repression between Glucose and Toluene Metabolism in Pseudomonas putida Is Channeled through Different Signaling Pathways

2007 ◽  
Vol 189 (18) ◽  
pp. 6602-6610 ◽  
Author(s):  
Teresa del Castillo ◽  
Juan L. Ramos
Keyword(s):  
Glucose Metabolism ◽  
Pseudomonas Putida ◽  
Catabolite Repression ◽  
Carbon Sources ◽  
Flux Analysis ◽  
Glucose Determination ◽  
Net Flux ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Metabolism Gene

ABSTRACT Pseudomonas putida KT2440(pWW0) can use toluene via the TOL plasmid-encoded catabolic pathways and can use glucose via a series of three peripheral chromosome-encoded routes that convert glucose into 6-phosphogluconate (6PG), namely, the glucokinase pathway, in which glucose is transformed to 6PG through the action of glucokinase and glucose-6-phosphate dehydrogenase. Alternatively, glucose can be oxidized to gluconate, which can be phosphorylated by gluconokinase to 6PG or oxidized to 2-ketogluconate, which, in turn, is converted into 6PG. Our results show that KT2440 metabolizes glucose and toluene simultaneously, as revealed by net flux analysis of [13C]glucose. Determination of glucokinase and gluconokinase activities in glucose metabolism, gene expression assays using a fusion of the promoter of the Pu TOL upper pathway to ′lacZ, and global transcriptomic assays revealed simultaneous catabolite repression in the use of these two carbon sources. The effect of toluene on glucose metabolism was directed to the glucokinase branch and did not affect gluconate metabolism. Catabolite repression of the glucokinase pathway and the TOL pathway was triggered by two different catabolite repression systems. Expression from Pu was repressed mainly via PtsN in response to high levels of 2-dehydro-3-deoxygluconate-6-phosphate, whereas repression of the glucokinase pathway was channeled through Crc.

scholarly journals Hierarchical management of carbon sources is regulated similarly by the CbrA/B systems in Pseudomonas aeruginosa and Pseudomonas putida

Microbiology ◽  
2014 ◽  
Vol 160 (10) ◽  
pp. 2243-2252 ◽  
Author(s):  
Martina Valentini ◽  
Sofía M. García-Mauriño ◽  
Isabel Pérez-Martínez ◽  
Eduardo Santero ◽  
Inés Canosa ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Pseudomonas Putida ◽  
Nitrogen Availability ◽  
Carbon Sources ◽  
Signal Transduction Pathway ◽  
Exact Nature ◽  
Energy Status ◽  
Regulatory Systems ◽  
Close Relationship ◽  
Carbon Nitrogen Ratio

The CbrA/B system in pseudomonads is involved in the utilization of carbon sources and carbon catabolite repression (CCR) through the activation of the small RNAs crcZ in Pseudomonas aeruginosa, and crcZ and crcY in Pseudomonas putida. Interestingly, previous works reported that the CbrA/B system activity in P. aeruginosa PAO1 and P. putida KT2442 responded differently to the presence of different carbon sources, thus raising the question of the exact nature of the signal(s) detected by CbrA. Here, we demonstrated that the CbrA/B/CrcZ(Y) signal transduction pathway is similarly activated in the two Pseudomonas species. We show that the CbrA sensor kinase is fully interchangeable between the two species and, moreover, responds similarly to the presence of different carbon sources. In addition, a metabolomics analysis supported the hypothesis that CCR responds to the internal energy status of the cell, as the internal carbon/nitrogen ratio seems to determine CCR and non-CCR conditions. The strong difference found in the 2-oxoglutarate/glutamine ratio between CCR and non-CCR conditions points to the close relationship between carbon and nitrogen availability, or the relationship between the CbrA/B and NtrB/C systems, suggesting that both regulatory systems sense the same sort or interrelated signal.

scholarly journals Levels and Activity of the Pseudomonas putida Global Regulatory Protein Crc Vary According to Growth Conditions

2005 ◽  
Vol 187 (11) ◽  
pp. 3678-3686 ◽  
Author(s):  
Ana Ruiz-Manzano ◽  
Luis Yuste ◽  
Fernando Rojo
Keyword(s):  
Pseudomonas Putida ◽  
Regulatory Protein ◽  
Carbon Sources ◽  
Signal Transduction Pathway ◽  
Growth Conditions ◽  
Degradation Pathway ◽  
Complete Medium ◽  
Alkane Degradation ◽  
Mutant Proteins ◽  
Catalytic Sites

ABSTRACT The global regulatory protein Crc is involved in the repression of several catabolic pathways for sugars, hydrocarbons, and nitrogenated and aromatic compounds in Pseudomonas putida and Pseudomonas aeruginosa when other preferred carbon sources are present in the culture medium (catabolite repression), therefore modulating carbon metabolism. We have analyzed whether the levels or the activity of Crc is regulated. Crc activity was followed by its ability to inhibit the induction by alkanes of the P. putida OCT plasmid alkane degradation pathway when cells grow in a complete medium, where the effect of Crc is very strong. The abundance of crc transcripts and the amounts of Crc protein were higher under repressing conditions than under nonrepressing conditions. The presence of crc on a high-copy-number plasmid considerably increased Crc levels, but this impaired its ability to inhibit the alkane degradation pathway. Crc shows similarity to a family of nucleases that have highly conserved residues at their catalytic sites. Mutation of the corresponding residues in Crc (Asp220 and His246) led to proteins that can inhibit induction of the alkane degradation pathway when present at normal or elevated levels in the cell. Repression by these mutant proteins occurred only under repressing conditions. These results suggest that both the amounts and the activity of Crc are modulated and support previous proposals that Crc may form part of a signal transduction pathway. Furthermore, the activity of the mutant proteins suggests that Crc is not a nuclease.

scholarly journals The Target for the Pseudomonas putida Crc Global Regulator in the Benzoate Degradation Pathway Is the BenR Transcriptional Regulator

2007 ◽  
Vol 190 (5) ◽  
pp. 1539-1545 ◽  
Author(s):  
Renata Moreno ◽  
Fernando Rojo
Keyword(s):  
Pseudomonas Putida ◽  
Target Genes ◽  
Carbon Sources ◽  
Degradation Pathway ◽  
Complete Medium ◽  
Mrna Levels ◽  
Global Regulator ◽  
Benzoate Degradation ◽  
Transcriptional Units ◽  
Degradation Intermediates

ABSTRACT Crc protein is a global regulator involved in catabolite repression control of several pathways for the assimilation of carbon sources in pseudomonads when other preferred substrates are present. In Pseudomonas putida cells growing exponentially in a complete medium containing benzoate, Crc strongly inhibits the expression of the benzoate degradation genes. These genes are organized into several transcriptional units. We show that Crc directly inhibits the expression of the peripheral genes that transform benzoate into catechol (the ben genes) but that its effect on genes corresponding to further steps of the pathway (the cat and pca genes of the central catechol and β-ketoadipate pathways) is indirect, since these genes are not induced because the degradation intermediates, which act as inducers, are not produced. Crc inhibits the translation of target genes by binding to mRNA. The expression of the ben, cat, and pca genes requires the BenR, CatR, and PcaR transcriptional activators, respectively. Crc significantly reduced benABCD mRNA levels but did not affect those of benR. Crc bound to the 5′ end of benR mRNA but not to equivalent regions of catR and pcaR mRNAs. A translational fusion of the benR and lacZ genes was sensitive to Crc, but a transcriptional fusion was not. We propose that Crc acts by reducing the translation of benR mRNA, decreasing BenR levels below those required for the full expression of the benABCD genes. This strategy provides great metabolic flexibility, allowing the hierarchical assimilation of different structurally related compounds that share a common central pathway by selectively regulating the entry of each substrate into the central pathway.

scholarly journals The Pseudomonas putida Crc Global Regulator Controls the Expression of Genes from Several Chromosomal Catabolic Pathways for Aromatic Compounds

2004 ◽  
Vol 186 (5) ◽  
pp. 1337-1344 ◽  
Author(s):  
Gracia Morales ◽  
Juan Francisco Linares ◽  
Ana Beloso ◽  
Juan Pablo Albar ◽  
José Luis Martínez ◽  
...  
Keyword(s):  
Pseudomonas Putida ◽  
Carbon Metabolism ◽  
Aromatic Compounds ◽  
Carbon Sources ◽  
Signal Transduction Pathway ◽  
Catabolic Repression ◽  
Catabolic Pathways ◽  
Expression Of Genes ◽  
Proteome Profile

ABSTRACT The Crc protein is involved in the repression of several catabolic pathways for the assimilation of some sugars, nitrogenated compounds, and hydrocarbons in Pseudomonas putida and Pseudomonas aeruginosa when other preferred carbon sources are present in the culture medium (catabolic repression). Crc appears to be a component of a signal transduction pathway modulating carbon metabolism in pseudomonads, although its mode of action is unknown. To better understand the role of Crc, the proteome profile of two otherwise isogenic P. putida strains containing either a wild-type or an inactivated crc allele was compared. The results showed that Crc is involved in the catabolic repression of the hpd and hmgA genes from the homogentisate pathway, one of the central catabolic pathways for aromatic compounds that is used to assimilate intermediates derived from the oxidation of phenylalanine, tyrosine, and several aromatic hydrocarbons. This led us to analyze whether Crc also regulates the expression of the other central catabolic pathways for aromatic compounds present in P. putida. It was found that genes required to assimilate benzoate through the catechol pathway (benA and catBCA) and 4-OH-benzoate through the protocatechuate pathway (pobA and pcaHG) are also negatively modulated by Crc. However, the pathway for phenylacetate appeared to be unaffected by Crc. These results expand the influence of Crc to pathways used to assimilate several aromatic compounds, which highlights its importance as a master regulator of carbon metabolism in P. putida.

scholarly journals Nitrobenzoates and Aminobenzoates Are Chemoattractants for Pseudomonas Strains

2004 ◽  
Vol 70 (1) ◽  
pp. 285-292 ◽  
Author(s):  
Rebecca E. Parales
Keyword(s):  
Pseudomonas Putida ◽  
Aromatic Compounds ◽  
Carbon Sources ◽  
Chemotactic Response ◽  
Aromatic Acids ◽  
Wide Range ◽  
Catechol 1,2 Dioxygenase ◽  
Sense And Respond ◽  
Chemotaxis System ◽  
Broad Specificity

ABSTRACT Three Pseudomonas strains were tested for the ability to sense and respond to nitrobenzoate and aminobenzoate isomers in chemotaxis assays. Pseudomonas putida PRS2000, a strain that grows on benzoate and 4-hydroxybenzoate by using the β-ketoadipate pathway, has a well-characterized β-ketoadipate-inducible chemotactic response to aromatic acids. PRS2000 was chemotactic to 3- and 4-nitrobenzoate and all three isomers of aminobenzoate when grown under conditions that induce the benzoate chemotactic response. P. putida TW3 and Pseudomonas sp. strain 4NT grow on 4-nitrotoluene and 4-nitrobenzoate by using the ortho (β-ketoadipate) and meta pathways, respectively, to complete the degradation of protocatechuate derived from 4-nitrotoluene and 4-nitrobenzoate. However, based on results of catechol 1,2-dioxygenase and catechol 2,3-dioxygenase assays, both strains were found to use the β-ketoadipate pathway for the degradation of benzoate. Both strains were chemotactic to benzoate, 3- and 4-nitrobenzoate, and all three aminobenzoate isomers after growth with benzoate but not succinate. Strain TW3 was chemotactic to the same set of aromatic compounds after growth with 4-nitrotoluene or 4-nitrobenzoate. In contrast, strain 4NT did not respond to any aromatic acids when grown with 4-nitrotoluene or 4-nitrobenzoate, apparently because these substrates are not metabolized to the inducer (β-ketoadipate) of the chemotaxis system. The results suggest that strains TW3 and 4NT have a β-ketoadipate-inducible chemotaxis system that responds to a wide range of aromatic acids and is quite similar to that present in PRS2000. The broad specificity of this chemotaxis system works as an advantage in strains TW3 and 4NT because it functions to detect diverse carbon sources, including 4-nitrobenzoate.

scholarly journals Growth independent rhamnolipid production from glucose using the non-pathogenic Pseudomonas putida KT2440

2011 ◽  
Vol 10 (1) ◽  
pp. 80 ◽  
Author(s):  
Andreas Wittgens ◽  
Till Tiso ◽  
Torsten T Arndt ◽  
Pamela Wenk ◽  
Johannes Hemmerich ◽  
...  
Keyword(s):  
Pseudomonas Putida ◽  
Pseudomonas Putida Kt2440 ◽  
Rhamnolipid Production

scholarly journals Crc Is Involved in Catabolite Repression Control of the bkd Operons of Pseudomonas putida andPseudomonas aeruginosa

2000 ◽  
Vol 182 (4) ◽  
pp. 1144-1149 ◽  
Author(s):  
Kathryn L. Hester ◽  
Jodi Lehman ◽  
Fares Najar ◽  
Lin Song ◽  
Bruce A. Roe ◽  
...  
Keyword(s):  
Pseudomonas Putida ◽  
Catabolite Repression ◽  
Carbon Sources ◽  
Keto Acid ◽  
Carbon Regulation ◽  
Acid Dehydrogenase ◽  
Transposon Mutants ◽  
Branched Chain ◽  
Glucose 6 Phosphate Dehydrogenase

ABSTRACT Crc (catabolite repression control) protein of Pseudomonas aeruginosa has shown to be involved in carbon regulation of several pathways. In this study, the role of Crc in catabolite repression control has been studied in Pseudomonas putida. The bkd operons of P. putida and P. aeruginosa encode the inducible multienzyme complex branched-chain keto acid dehydrogenase, which is regulated in both species by catabolite repression. We report here that this effect is mediated in both species by Crc. A 13-kb cloned DNA fragment containing the P. putida crc gene region was sequenced. Crc regulates the expression of branched-chain keto acid dehydrogenase, glucose-6-phosphate dehydrogenase, and amidase in both species but not urocanase, although the carbon sources responsible for catabolite repression in the two species differ. Transposon mutants affected in their expression of BkdR, the transcriptional activator of thebkd operon, were isolated and identified as crcand vacB (rnr) mutants. These mutants suggested that catabolite repression in pseudomonads might, in part, involve control of BkdR levels.

Responses of Pseudomonas putida to toxic aromatic carbon sources

2012 ◽  
Vol 160 (1-2) ◽  
pp. 25-32 ◽  
Author(s):  
Tino Krell ◽  
Jesús Lacal ◽  
M. Eugenia Guazzaroni ◽  
Andreas Busch ◽  
Hortencia Silva-Jiménez ◽  
...  
Keyword(s):  
Pseudomonas Putida ◽  
Carbon Sources ◽  
Aromatic Carbon
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