Characterisation of rhamnolipid production in a previously un-investigated, non-pathogenic marine Pseudomonad.

Matthew Twigg, Lakshmi Tripathi, Katarina Zompra, Karina Salek, Victor Irorere, Tony Gutierrez, George Spyroulias, Roger Marchant, Ibrahim Banat

Research output: Contribution to conferencePoster

Abstract

This study identifies, for the first time a significantly high level of synthesis of short chain di-rhamnolipids by a non-pathogenic marine Pseudomonas species. Pseudomonas sp. MCTG214(3b1) was isolated from a phytoplankton bloom of the coast of Florida, phylogenetically identified using 16S rDNA and gyrB sequencing, however was found to be unrelated to P. aeruginosa. Cell-free supernatant samples obtained from this strain showed significant reductions in surface tension signifying potential biosurfactant synthesis. HPLC-MS and NMR analysis of these samples indicated the presence of five different rhamnolipid congeners. Di-rhamnolipid synthesis accounted for 87% relative abundance and all congeners possessed fatty acid moieties consisting of 8-12 carbons. PCR screening of this strain revealed homologues to the P. aeruginosa RL synthesis genes rhlA and rhlB, however no rhlC homologue was identified. Using the Galleria mellonella larvae model, the strain was demonstrated to be far less pathogenic than P. aeruginosa. We postulate that RL synthesis in Pseudomonas sp. MCTG214(3b1) is carried out by enzymes expressed from rhlA/B homologues similar to those of P. aeruginosa, however a lack of rhlC potentially indicates the presence of an alternative yet unknown novel rhamnosyltransferase responsible for the production of the di-rhamnolipid congeners identified by HPLC-MS.
LanguageEnglish
Number of pages1
Publication statusPublished - 24 Sep 2019

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surface tension
nuclear magnetic resonance
relative abundance
algal bloom
fatty acid
phytoplankton
enzyme
larva
gene
coast
carbon
analysis
screening

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Twigg, Matthew ; Tripathi, Lakshmi ; Zompra, Katarina ; Salek, Karina ; Irorere, Victor ; Gutierrez, Tony ; Spyroulias, George ; Marchant, Roger ; Banat, Ibrahim. / Characterisation of rhamnolipid production in a previously un-investigated, non-pathogenic marine Pseudomonad. 1 p.
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title = "Characterisation of rhamnolipid production in a previously un-investigated, non-pathogenic marine Pseudomonad.",
abstract = "This study identifies, for the first time a significantly high level of synthesis of short chain di-rhamnolipids by a non-pathogenic marine Pseudomonas species. Pseudomonas sp. MCTG214(3b1) was isolated from a phytoplankton bloom of the coast of Florida, phylogenetically identified using 16S rDNA and gyrB sequencing, however was found to be unrelated to P. aeruginosa. Cell-free supernatant samples obtained from this strain showed significant reductions in surface tension signifying potential biosurfactant synthesis. HPLC-MS and NMR analysis of these samples indicated the presence of five different rhamnolipid congeners. Di-rhamnolipid synthesis accounted for 87{\%} relative abundance and all congeners possessed fatty acid moieties consisting of 8-12 carbons. PCR screening of this strain revealed homologues to the P. aeruginosa RL synthesis genes rhlA and rhlB, however no rhlC homologue was identified. Using the Galleria mellonella larvae model, the strain was demonstrated to be far less pathogenic than P. aeruginosa. We postulate that RL synthesis in Pseudomonas sp. MCTG214(3b1) is carried out by enzymes expressed from rhlA/B homologues similar to those of P. aeruginosa, however a lack of rhlC potentially indicates the presence of an alternative yet unknown novel rhamnosyltransferase responsible for the production of the di-rhamnolipid congeners identified by HPLC-MS.",
author = "Matthew Twigg and Lakshmi Tripathi and Katarina Zompra and Karina Salek and Victor Irorere and Tony Gutierrez and George Spyroulias and Roger Marchant and Ibrahim Banat",
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Characterisation of rhamnolipid production in a previously un-investigated, non-pathogenic marine Pseudomonad. / Twigg, Matthew; Tripathi, Lakshmi; Zompra, Katarina; Salek, Karina; Irorere, Victor; Gutierrez, Tony; Spyroulias, George ; Marchant, Roger; Banat, Ibrahim.

2019.

Research output: Contribution to conferencePoster

TY - CONF

T1 - Characterisation of rhamnolipid production in a previously un-investigated, non-pathogenic marine Pseudomonad.

AU - Twigg, Matthew

AU - Tripathi, Lakshmi

AU - Zompra, Katarina

AU - Salek, Karina

AU - Irorere, Victor

AU - Gutierrez, Tony

AU - Spyroulias, George

AU - Marchant, Roger

AU - Banat, Ibrahim

PY - 2019/9/24

Y1 - 2019/9/24

N2 - This study identifies, for the first time a significantly high level of synthesis of short chain di-rhamnolipids by a non-pathogenic marine Pseudomonas species. Pseudomonas sp. MCTG214(3b1) was isolated from a phytoplankton bloom of the coast of Florida, phylogenetically identified using 16S rDNA and gyrB sequencing, however was found to be unrelated to P. aeruginosa. Cell-free supernatant samples obtained from this strain showed significant reductions in surface tension signifying potential biosurfactant synthesis. HPLC-MS and NMR analysis of these samples indicated the presence of five different rhamnolipid congeners. Di-rhamnolipid synthesis accounted for 87% relative abundance and all congeners possessed fatty acid moieties consisting of 8-12 carbons. PCR screening of this strain revealed homologues to the P. aeruginosa RL synthesis genes rhlA and rhlB, however no rhlC homologue was identified. Using the Galleria mellonella larvae model, the strain was demonstrated to be far less pathogenic than P. aeruginosa. We postulate that RL synthesis in Pseudomonas sp. MCTG214(3b1) is carried out by enzymes expressed from rhlA/B homologues similar to those of P. aeruginosa, however a lack of rhlC potentially indicates the presence of an alternative yet unknown novel rhamnosyltransferase responsible for the production of the di-rhamnolipid congeners identified by HPLC-MS.

AB - This study identifies, for the first time a significantly high level of synthesis of short chain di-rhamnolipids by a non-pathogenic marine Pseudomonas species. Pseudomonas sp. MCTG214(3b1) was isolated from a phytoplankton bloom of the coast of Florida, phylogenetically identified using 16S rDNA and gyrB sequencing, however was found to be unrelated to P. aeruginosa. Cell-free supernatant samples obtained from this strain showed significant reductions in surface tension signifying potential biosurfactant synthesis. HPLC-MS and NMR analysis of these samples indicated the presence of five different rhamnolipid congeners. Di-rhamnolipid synthesis accounted for 87% relative abundance and all congeners possessed fatty acid moieties consisting of 8-12 carbons. PCR screening of this strain revealed homologues to the P. aeruginosa RL synthesis genes rhlA and rhlB, however no rhlC homologue was identified. Using the Galleria mellonella larvae model, the strain was demonstrated to be far less pathogenic than P. aeruginosa. We postulate that RL synthesis in Pseudomonas sp. MCTG214(3b1) is carried out by enzymes expressed from rhlA/B homologues similar to those of P. aeruginosa, however a lack of rhlC potentially indicates the presence of an alternative yet unknown novel rhamnosyltransferase responsible for the production of the di-rhamnolipid congeners identified by HPLC-MS.

M3 - Poster

ER -