Dirhamnose-lipid production by recombinant nonpathogenic bacterium Pseudomonas chlororaphis

Solaiman, Daniel K. Y.; Ashby, Richard D.; Gunther, Nereus W.; Zerkowski, Jonathan A.

doi:10.1007/s00253-015-6433-4

Cited by 29 publications

(15 citation statements)

References 50 publications

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“…Finally, we included the rhlA from P. fluorescens LMG 05825 (P. chlororaphis ATCC 17813), which is reported to be the same strain as P. chlororaphis NRRL B-30761 (35), a strain producing mainly C 10 -C 12 and C 10 -C 12:1 congeners (15). Solaiman et al (36) found an operon containing rhlAB and the regulator gene rhlR (Fig. 3) in this strain.…”

Section: Figsupporting

confidence: 54%

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Exploiting the Natural Diversity of RhlA Acyltransferases for the Synthesis of the Rhamnolipid Precursor 3-(3-Hydroxyalkanoyloxy)Alkanoic Acid

Germer

Tiso

Müller

et al. 2020

Appl Environ Microbiol

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While rhamnolipids of the Pseudomonas aeruginosa type are commercially available, the natural diversity of rhamnolipids and their origin have barely been investigated. Here, we collected known and identified new rhlA genes encoding the acyltransferase responsible for the synthesis of the lipophilic rhamnolipid precursor 3-(3-hydroxyalkanoyloxy)alkanoic acid (HAA). Generally, all homologs were found in Betaproteobacteria and Gammaproteobacteria. A likely horizontal gene transfer event into Actinobacteria is the only identified exception. The phylogeny of the RhlA homologs from Pseudomonas and Burkholderia species is consistent with the organism phylogeny, and genes involved in rhamnolipid synthesis are located in operons. In contrast, RhlA homologs from the Enterobacterales do not follow the organisms’ phylogeny but form their own branch. Furthermore, in many Enterobacterales and Halomonas from the Oceanospirillales, an isolated rhlA homolog can be found in the genome. The RhlAs from Pseudomonas aeruginosa PA01, Pseudomonas fluorescens LMG 05825, Pantoea ananatis LMG 20103, Burkholderia plantarii PG1, Burkholderia ambifaria LMG 19182, Halomonas sp. strain R57-5, Dickeya dadantii Ech586, and Serratia plymuthica PRI-2C were expressed in Escherichia coli and tested for HAA production. Indeed, except for the Serratia RhlA, HAAs were produced with the engineered strains. A detailed analysis of the produced HAA congeners by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) highlights the congener specificity of the RhlA proteins. The congener length varies from 4 to 18 carbon atoms, with the main congeners consisting of different combinations of saturated or monounsaturated C10, C12, and C14 fatty acids. The results are discussed in the context of the phylogeny of this unusual enzymatic activity. IMPORTANCE The RhlA specificity explains the observed differences in 3-(3-hydroxyalkanoyloxy)alkanoic acid (HAA) congeners. Whole-cell catalysts can now be designed for the synthesis of different congener mixtures of HAAs and rhamnolipids, thereby contributing to the envisaged synthesis of designer HAAs.

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Section: Figsupporting

confidence: 54%

“…the C 12 chain is unsaturated in one position (15,35,36). In contrast, Burkholderia kururiensis KP23 produces Gammaproteobacteria-like rhamnolipids containing mainly C 10 -C 10 residues (24).…”

mentioning

confidence: 99%

Exploiting the Natural Diversity of RhlA Acyltransferases for the Synthesis of the Rhamnolipid Precursor 3-(3-Hydroxyalkanoyloxy)Alkanoic Acid

Germer

Tiso

Müller

et al. 2020

Appl Environ Microbiol

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“…The authors reported that the modified P. chlororaphis was able to produce 2.4 fold more dirhamnolipids than mono-rhamnolipids [76]. A previous study of the same group reported that a 3-hydroxyacyl-ACP/CoA transacylase (phaG)-knockout does not affect rhamnolipid synthesis [77].…”

Section: P Aeruginosa Genes In Heterologous Hostsmentioning

confidence: 96%

“…In these ways, a modified P. chlororaphis strain was also designed, harboring plasmid pBS29-P2-rhlC, and expressing of the rhamnosyltransferase C (rhlC) gene from P. aeruginosa, under control of a Pseudomonas syringae promoter [76]. The authors reported that the modified P. chlororaphis was able to produce 2.4 fold more dirhamnolipids than mono-rhamnolipids [76].…”

Section: P Aeruginosa Genes In Heterologous Hostsmentioning

confidence: 98%

Rhamnolipids in perspective: gene regulatory pathways, metabolic engineering, production and technological forecasting

et al. 2016

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“…and Burkholderia sp., implying that the metabolic engineering can also provide a means in modifying the composition and types of rhamnolipid congeners produced to alter the physicochemical properties of the rhamnolipids [ 48 ]. Likewise, P. chlororaphis, which was previously reported to produce only mono-rhamnolipids, was shown to be able to produce di-rhamnolipids by overexpressing rhlC under a constitutive promoter [ 49 ]. The rhamnolipid composition was also altered by site-directed mutagenesis on rhlB that resulted in the modification to the volume of the substrate binding pocket [ 50 ].…”

Section: Application Of Various Strategies To Increase Rhamnolipid Yimentioning

confidence: 99%

Microbial production of rhamnolipids: opportunities, challenges and strategies

2017

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Rhamnolipids are a class of biosurfactants which contain rhamnose as the sugar moiety linked to β-hydroxylated fatty acid chains. Rhamnolipids can be widely applied in many industries including petroleum, food, agriculture and bioremediation etc. Pseudomonas aeruginosa is still the most competent producer of rhamnolipids, but its pathogenicity may cause safety and health concerns during large-scale production and applications. Therefore, extensive studies have been carried out to explore safe and economical methods to produce rhamnolipids. Various metabolic engineering efforts have also been applied to either P. aeruginosa for improving its rhamnolipid production and diminishing its pathogenicity, or to other non-pathogenic strains by introducing the key genes for safe production of rhamnolipids. The three key enzymes for rhamnolipid biosynthesis, RhlA, RhlB and RhlC, are found almost exclusively in Pseudomonas sp. and Burkholderia sp., but have been successfully expressed in several non-pathogenic host bacteria to produce rhamnolipids in large scales. The composition of mono- and di-rhamnolipids can also be modified through altering the expression levels of RhlB and RhlC. In addition, cell-free rhamnolipid synthesis by using the key enzymes and precursors from non-pathogenic sources is thought to not only eliminate pathogenic effects and simplify the downstream purification processes, but also to circumvent the complexity of quorum sensing system that regulates rhamnolipid biosynthesis. The pathogenicity of P. aeruginosa can also be reduced or eliminated through in vivo or in vitro enzymatic degradation of the toxins such as pyocyanin during rhamnolipid production. The rhamnolipid production cost can also be significantly reduced if rhamnolipid purification step can be bypassed, such as utilizing the fermentation broth or the rhamnolipid-producing strains directly in the industrial applications of rhamnolipids.

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Dirhamnose-lipid production by recombinant nonpathogenic bacterium Pseudomonas chlororaphis

Cited by 29 publications

References 50 publications

Exploiting the Natural Diversity of RhlA Acyltransferases for the Synthesis of the Rhamnolipid Precursor 3-(3-Hydroxyalkanoyloxy)Alkanoic Acid

Exploiting the Natural Diversity of RhlA Acyltransferases for the Synthesis of the Rhamnolipid Precursor 3-(3-Hydroxyalkanoyloxy)Alkanoic Acid

Rhamnolipids in perspective: gene regulatory pathways, metabolic engineering, production and technological forecasting

Microbial production of rhamnolipids: opportunities, challenges and strategies

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