Methods for Recombinant Rhamnolipid Production

Tiso, Till; Germer, Andrea; Küpper, Benjamin; Wichmann, R.; Blank, Lars M.

doi:10.1007/8623_2015_60

Cited by 7 publications

(8 citation statements)

References 36 publications

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“…Quantification of HAAs. For HAA quantification, components of the samples were separated using reverse-phase chromatography on a C 18 column and detected with a charged aerosol detector (RP-HPLC-CAD) as described by Tiso et al (60), based on the method established by Behrens et al (61). Harvesting of cells and cell residue was accomplished by centrifugation at 13,000 ϫ g for 3 min.…”

Section: Discussionmentioning

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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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: Discussionmentioning

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 natural RL producer P. aeruginosa is an opportunistic human pathogen entailing a complex regulation, which limits industrial application (Gunther et al, 2005). To overcome this restriction, the safety‐level‐1‐organism P. putida KT2440, which is an industrially relevant microorganism, was genetically modified to produce RL by introducing two required genes from P. aeruginosa (Tiso et al, 2015, 2017; Wittgens et al, 2011). RL production from hydrophilic substrates like glucose or xylose with P. putida was shown before (Bator et al, 2020; Tiso et al, 2020), drastically improving the downstream process in contrast to hydrophobic substrates like plant oils, which are often used for RL synthesis by P. aeruginosa (Wittgens et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

A scalable bubble‐free membrane aerator for biosurfactant production

Bongartz

Bator

Baitalow

et al. 2021

Biotech & Bioengineering

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The bioeconomy is a paramount pillar in the mitigation of greenhouse gas emissions and climate change. Still, the industrialization of bioprocesses is limited by economical and technical obstacles. The synthesis of biosurfactants as advanced substitutes for crude‐oil‐based surfactants is often restrained by excessive foaming. We present the synergistic combination of simulations and experiments towards a reactor design of a submerged membrane module for the efficient bubble‐free aeration of bioreactors. A digital twin of the combined bioreactor and membrane aeration module was created and the membrane arrangement was optimized in computational fluid dynamics studies with respect to fluid mixing. The optimized design was prototyped and tested in whole‐cell biocatalysis to produce rhamnolipid biosurfactants from sugars. Without any foam formation, the new design enables a considerable higher space–time yield compared to previous studies with membrane modules. The design approach of this study is of generic nature beyond rhamnolipid production.

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“…Not surprisingly, the low-hanging fruits are to be found in the realm of metabolic engineering, a field that is well familiar with the use of bacteria other than E. coli as platforms for synthesis of added-value products. The panel of cases examined below go from biosynthesis of bulk chemicals such as isoprenoids [29], esters [30] and rhamnolipids [31], to biotransformations for adding value to common substrates [32] and finally to produce more sophisticated molecules like lantibiotics [33]. New materials with awesome properties can also be produced by deeply refactored bacteria [34].…”

Section: Discussionmentioning

confidence: 99%

Introduction to Systems and Synthetic Biology in Hydrocarbon Microbiology: Applications

Lorenzo

2016

Springer Protocols Handbooks

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Contemporary Microbial Biotechnology is experiencing a rapid transition between being a mostly trial-anderror endeavour towards becoming a quantitative and predictable branch of contemporary research. A key ingredient of this shift involves the adoption of Systems and Synthetic Biology approaches for either revisiting typical themes (e.g. bioproduction of added-value molecules) or to develop altogether new ones (such as engineering of sensor/actuator devices). The first wave of goods reaching the biotechnological sector largely comes from metabolic engineering of microorganisms for biofuels, fine chemicals and high-added value molecules. But much more is still to come by applying electric and industrial engineering principles to biological systems as well as by learning from the way natural evolution has solved apparently intractable design problems.

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Methods for Recombinant Rhamnolipid Production

Cited by 7 publications

References 36 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

A scalable bubble‐free membrane aerator for biosurfactant production

Introduction to Systems and Synthetic Biology in Hydrocarbon Microbiology: Applications

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