List of Oral/Poster Presentations
| Date | Presentation (An * is added on the name of the presenter) | |||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 2024-03 | A journey in the optimization of CRISPR-Cas9-based genome editing tools for Yarrowia lipolytica | Oral Presentation | Invited | Synthetic Biology Club, Abdel-Mawgoud Synthetic Biology Lab, Université Laval, Québec, Canada | Abdel-Mawgoud SB club | https://abdel-mawgoud.com/en/presentation-entry-form/ | Yarrowia lipolytica is an important oleaginous yeast used in the production of specialty chemicals and has a great potential for further applications in lipid biotechnology. Harnessing the full potential of Y. lipolytica is, however, limited by its inherent recalcitrance to traditional genetic manipulations. Current CRISPR/Cas9-based methods resulted in great advances in genome editing of this yeast, yet it had limitations in terms of the number of editable genomic targets as well as in the efficiency at which they can be edited. We further improved CRISPR-Cas9 editing protocol by optimising the expression cassettes of gRNA and Cas9. For optimisation of gRNA expression, we used tools of secondary RNA structure prediction that guided our redesign of the tRNA-sgRNA fusions used for the expression of single guide RNA (sgRNA). This resulted in high efficiency CRISPR/cas9 genome editing at chromosomal loci that failed gene editing or were previously editable at very low efficiencies and using templates with short homology arms. For optimisation of Cas9 expression, we used tools of prediction of promoter elements that allowed the reengineering of a short TEF promoter 5-fold higher gene expression than currently engineered TEF promoters. Expression of Cas9 under the reengineered short TEF promoter increased gene integration efficiency. These optimized genome editing tools are expected to enhance the genetic and metabolic engineering of the yeast, Yarrowia lipolytica, for various industrial purposes. | A. M. Abdel-Mawgoud* | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-064664978e31d3945754bddcc8c94f2d-ff-2024-03-25-CRISPR-Cas9-in-YALI-Ahmad-Saleh.jpg | No | ||||||
| 2021-05 | Komagataeibacter rhaeticus, une productrice intéressante de cellulose, mais limitée à cause des éléments transposables | Poster Presentation | Congrès de Bactériologie intégrative: Symbiose - Pathogenèse, Online, Québec, Canada | Congrès BiSP 2021 | https://event.fourwaves.com/fr/bisp2021/pages | https://event.fourwaves.com/fr/bisp2021/resumes/19ce980b-8cd7-47ed-a34b-416d2c63e24c | La cellulose d’origine végétale est le polymère biologique le plus commun dans notre biosphère, et l’humain en fait déjà usage sous la forme de bois, papier, coton, et autres matériaux. Cependant, la cellulose bactérienne possède des propriétés plus intéressantes que la cellulose végétale pour une pléthore de domaines, qui vont de la médecine (pansements de qualité et greffons biocompatibles) à l’électronique (écrans flexibles). Celle-ci est produite sous la forme de membranes à la surface des cultures de certaines bactéries de la famille de Acetobacteraceae, dont la meilleure productrice est Komagataeibacter rhaeticus.Cependant, la production de cellulose par K. rhaeticus rencontre des problèmes importants, dont l’instabilité génétique causée par des éléments transposables ainsi que le faible rendement de ce polymère. Pour mieux comprendre les facteurs jouant un rôle dans l’instabilité génétique, nous avons examiné le génome de K. rhaeticus et nous y avons trouvé plus de 17 transposases, dont nous avons défini les familles. Il fut intéressant de constater que la majorité des gènes impliqués dans la biosynthèse de cellulose sont infiltrés par des séquences d’insertion, ce qui explique l’instabilité génétique de cette bactérie est pourrait être relié à la variation au niveau de sa production de cellulose.Cette connaissance aidera à développer une stratégie de génie génétique et métabolique qui permettra une surproduction stable de cellulose à un niveau économiquement rentable chez cette bactérie. | B. Arcand*; J. Fréchette; Y. Messaddeq; A. M. Abdel-Mawgoud | https://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-c05f89fa4b1485ccfcefcf058517ef19-ff-BiSP-2021-Affiche-BA.jpg | no | ||||||
| 2021-11 | Development of tools for genetic engineering of cellulose production in Komagataeibacter | Poster Presentation | Réunion scientifique Sentinelle Nord 2021, Online, Québec, Canada | https://sentinellenord.ulaval.ca/fr/reunion-scientifique-2021 | Cellulose harvested from plants is the most common polymer of biological origin, and we already use it extensively as it is an important component of wood, cotton, paper, and other materials. However, bacterial cellulose, or BC, exhibits unique properties that make it especially suited for a plethora of applications, such as water filtration, high-quality bandages, and optoelectronics. BC is produced at the air/medium interface of liquid cultures of some bacteria belonging to the Acetobacteraceae family, mostly of the Komagataeibacter genus, and are easily harvested and processed.However, production of bacterial cellulose at an industrial scale meets important challenges, such as low yields and instability of cellulose production in culture, which translate to high production costs. This project aims to build a toolkit for the genetic engineering of cellulose-producing bacteria and use these to produce an optimized strain for industrial-scale BC production by addressing these challenges directly at their source.So far, we identified a suitable plasmid backbone, multiple selection markers and their resistance cassettes, and built a synthetic CRISPR-Cas9 gene codon-optimized for use in Komagataeibacter. Moreover, we measured the phenotypic instability of cellulose production in three Komagataeibacter strains and began trials for optimization of the growth medium.Additionally, we produced a library of cellulose-nonproducing mutants of Komagataeibacter rhaeticus iGEM and screened these for known alterations to the genes responsible for BC production, such as the presence of mutations and transposable elements.We expect this work will provide a roadmap for further experimentation and ultimately will make production of bacterial cellulose more viable from an economic standpoint. | B. Arcand*; J. Fréchette; Y. Messaddeq; A. M. Abdel-Mawgoud | https://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-4fe030e2f91523f2339480672ab8b0ea-ff-RS-SentinelleNord2021-BA.jpg | no | ||||||||
| 2023-09 | Identification d’un élément transposable interférant avec la production de cellulose chez Komagataeibacter rhaeticus, et remédiation par expression d’une séquence modifiée du gène cible | Poster Presentation | Congrès de Bactériologie intégrative: Symbiose – Pathogenèse, Pavillon Roger Gaudry, Université de Montréal, Québec, Canada. | Congrès BiSP 2023 | https://event.fourwaves.com/fr/bisp2023/pages | https://event.fourwaves.com/fr/bisp2023/resumes/932fbf2a-ba9e-4bcc-ba25-0bf157a5061d | La cellulose bactérienne possède un potentiel supérieur à celui de la cellulose végétale pour plusieurs domaines, qui vont de la médecine (pansements pour grands brûlés et greffons biocompatibles) à l’électronique (écrans flexibles) jusqu’à l’exploration spatiale. Celle-ci est produite sous la forme de membranes à la surface des cultures de certaines bactéries de la famille des Acetobacteraceae, dont la meilleure productrice est Komagataeibacter rhaeticus. Cependant, la production de cellulose par K. rhaeticus rencontre plusieurs défis, dont l’instabilitégénétique causée par des éléments transposables, laquelle limite le rendement cellulosique. Pour mieux comprendre les facteurs jouant un rôle dans cette instabilité génétique, nous avons examiné le génomede K. rhaeticus et nous y avons trouvé plus de 17 transposases.Nous avons ensuite confirmé par séquençage l’insertion de l’élément transposable IS1032 à un site précis dans la séquence du gène bcsA chez plusieurs mutants non-producteurs de cellulose. Une séquence mutée de ce gène fut assemblée sur un châssis plasmidique d’expression puis transformé dans un clone non-producteur de cellulose, entraînant la réversion à un phénotype de production de cellulose. | B. Arcand*; J. Fréchette; Y. Messaddeq; A. M. Abdel-Mawgoud | https://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-3123c9537c80f8606426c4de6197c911-ff-BiSP-2023-Affiche-BA.jpg | no | ||||||
| 2021-05 | Augmentation de l’expression de Cas9 pour la modification génétique efficace médiée par un CRISPR-Cas9 d’une souche de Yarrowia lipolytica surproductrice de lipide | Oral Presentation | Congrès de Bactériologie intégrative: Symbiose - Pathogenèse, Online, Québec, Canada | Congrès BiSP 2021 | https://event.fourwaves.com/fr/bisp2023/pages | La levure oléagineuse Yarrowia lipolytica, sous certaine condition, a la capacité d’accumuler de hauts titres de lipide (entre 40 et 70% de sa masse sèche). Ceci la considère comme un organisme potentiel en biotechnologie pour la production de dérivés lipidiques comme les biocarburants et les acides gras polyinsaturés. Pour manipuler le rendement et/ou la nature des lipides, il est nécessaire de procéder à l’édition du génome du châssis pour rediriger le flux métabolique vers les molécules d’intérêts. Nous démontrons que l’augmentation de l’expression du Cas9 par un promoteur fort augmente l’efficacité de la délétion des gènes par CRISPR-Cas9 médiée par un oligonucléotide de délétion de 100 bp spécifique au gène cible. Nous avons validé l’efficacité de la nouvelle méthode en comparant la délétion sur plusieurs gènes en utilisant un Cas9 sous un promoteur fort et un conventionnel. L’efficacité de délétion “Knock-out” a été analysée au niveau phénotypique et génotypique. Nous avons constaté plusieurs évènements génotypiques développés à cause de la mutation des gènes. Ces événements génotypiques ont été caractérisés par le séquençage des gènes délétés afin de comprendre les mécanismes sous-jacents aux différents événements de délétions. La méthode développée a été utilisée efficacement pour la construction d’une souche de Y. lipolytica sur productrice des lipides en délétant des gènes comme les gènes TGL3 et TGL4 initiant la voie de dégradation des triglycérides. Cette étude présente une méthode d’édition génomique efficace facilitant les manipulations génétiques et métaboliques pour mieux exploiter cette levure comme châssis industriel et biotechnologique. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-d5e6c0c19a369d367a83d0651605fe25-ff-OuelletB_BISP-2021-main.png | no | |||||||
| 2021-11 | Impact of modulating Cas9 expression on the efficiency of CRISPR-Cas9-mediate genome editing in Yarrowia lipolytica | Oral Presentation | Journée de la Recherche en Sciences et Génie (2021), Université Laval, Québec, Canada. | JRSG 2021 | CRISPR-Cas is a powerful tool for genome edition that is particularly needed nowadays for the genetic and metabolic engineering of industrial microorganisms like the oleaginous yeast Yarrowia lipolytica. This tool demonstrated a great success in the genetic manipulation of Y. lipolytica to harness it towards the production of, amongst others, essential polyunsaturated fatty acids and biodiesel. Recently, an efficient CRISPR-Cas9-mediated genome editing protocol was developed for Y. lipolytica, that is based on tRNA-sgRNA fusion and in which Cas9 is under the control of a synthetic promoter consisting of upstream-activating sequences in tandem fused to the TEF promoter (UAS1B8-TEF). However, tandem repeats in DNA sequences can lead to polymerase slippage and are difficult to manipulate for cloning purposes. Here we report a promoter composed of a truncated version of the native TEF promoter (TEFNative) of Y. lipolytica. The promoter strengths of the TEFNative (406 bp), the truncated TEF (TEFTrunc, 366 bp) and UAS1B8-TEF (1041 bsp) were compared using hrGFP fusions. Fluorescence readings revealed that TEFNative and TEFTrunc promoters have equivalent expression strengths, whereas the UAS1B8-TEF is significantly weaker. We also compared the CRISPR-Cas9-mediated genome editing efficiency and the associated genotypic and phenotypic modifications when using Cas9 under control of the 3 promoters. Interestingly, both TEFNative and TEFTrunc promoters have demonstrated equivalent efficiencies of CRISPR-Cas9-mediated genome editing on the genotypic and phenotypic levels. We are currently comparing these Cas9-editing efficiencies with that when using Cas9 under UAS1B8-TEF promoter. Here, we redesigned the Cas9 expression mechanism that guarantees an enhanced expression and genome editing efficiency and phenotype modification using a shorter and easily manipulable promoter, like TEFTrunc, than the one proposed in literature. These easy-to-use genetic parts will facilitate synthetic biology and genetic engineering of Y. lipolytica for the production of bulk and fine chemical. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-88b3f4bf0797e65a93ac2612a734f77b-ff-OuelletB_JRSG-2021-main.png | yes | Monetary | 1000 | ||||||
| 2021-11 | Impact of modulating Cas9 expression on the efficiency of CRISPR-Cas9-mediate genome editing in Yarrowia lipolytica | Oral Presentation | Invited | Synthetic Biology Club, Abdel-Mawgoud Synthetic Biology Lab, Université Laval, Québec, Canada | Abdel-Mawgoud SB Club | https://abdel-mawgoud.com/en/synthetic-biology-club/ | CRISPR-Cas is a powerful tool for genome editing. This tool demonstrated a great success in the genetic manipulation of Yarrowia lipolytica that opened the doors towards harness the lipogenic potential of this yeast towards the production of essential polyunsaturated fatty acids, biodiesel and other oleochemicals. To enhance protein expression, a synthetic hybrid promoter consisting of eight upstream-activating sequences in tandem fused to the TEF promoter was previously proposed and is currently used for the expression of Cas9 in CRISPR-Cas9-mediated genome editing protocols for Y. lipolytica. However, tandem repeats are difficult to manipulate or clone. Here we report a truncated version of the native TEF promoter of Y. lipolytica. Our results showed that this truncated TEF is a significantly stronger promoter than UAS1B8-TEF as estimated using GFP reporter genes. Comparison of genome editing using Cas9 under the two promoters revealed that the truncated TEF leads to higher gene deletion efficiency. We propose this new truncated TEF promoter to drive Cas9 expression to enhance its expression and hence its genome editing efficiency. Moreover, being shorter, truncated TEF is also easier to manipulate. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-a51c3d55bf91bf362159c181f9a941c8-ff-OuelletB_SBClub-2021-main.png | no | ||||||
| 2022-05 | Novel synthetic promoter for higher expression and CRISPR-Cas9-mediated genome editing efficiency in Yarrowia lipolytica | Oral Presentation | PROTEO Annual Symposium (2022), hybrid event, UQAM, Montreal, QC, Canada | https://proteo.ca/en/news-and-events/proteo-annual-symposium-2022/#:~:text=The2022PROTEOAnnualSymposiumwill | https://event.fourwaves.com/fr/proteo2022/resumes/e7c58a00-ae38-4337-a524-777a228e8fa4/video | The non-conventional yeast Yarrowia lipolytica is gaining interest in biotechnology for the production of recombinant protein and other biomolecules. However, site-specific genome editing is limited in Y. lipolytica. This was overcome by the development of a CRISPR-Cas9 genome editing protocol for Y. lipolytica based on a tRNA-sgRNA fusion and in which Cas9 is under the control of a synthetic promoter, UAS1B8-TEF136, previously reported as a strong one. It consisted of eight tandem upstream-activating sequences fused to a minimal TEF promoter (TEF136). However, tandem repeats in DNA sequences are prone to polymerase slippage and are difficult to manipulate for cloning purposes. Here we report another synthetic promoter composed of a truncated version (TEF366) of the native Y. lipolytica TEF promoter (TEF406). Assessment of the expression rate of the two synthetic promoters by GFP reporters revealed that TEF366 and TEF406 are associated with higher expression by more than 10 folds relative to UAS1B8-TEF136. We then compared genome editing efficiency using Cas9 under each of the three promoters. Cas9 under TEF366 and TEF406 promoters showed to be associated with higher chromosomal site-specific gene insertion by 40% compared to that under UAS1B8-TEF136 promoter. Furthermore, for site-specific genomic deletions, Cas9 under TEF366 or TEF406 showed no significant difference both on the genotypic and phenotypic levels. We conclude that the higher is the Cas9 expression, the higher is its genome editing efficiency in Y. lipolytica. In addition, the simple TEF-based promoters, native or truncated, are stronger than the hybrid UAS1B8-TEF136 promoter. Being shorter, the truncated TEF (TEF366) is more convenient than the complete TEF (TEF406). This makes TEF366 an easy-to-use synthetic biology part for genetic engineering of Y. lipolytica for production of recombinant proteins and other biochemicals. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-f058a795eb3a558ade4a6dcb36b238c1-ff-OuelletB_PROTEO-2022-main.png | yes | Non-monetary | ||||||
| 2023-05 | Expression of Cas9 under an engineered 3’- and 5’ truncated TEF1α promoter enhances CRISPR-Cas9-mediated genome editing in Yarrowia lipolytica | Poster Presentation | PROTEO Annual Symposium (2023), Théâtre de la Cité Universitaire (TCU), Palasis-Prince Hall, Laval University,Quebec, QC, Canada | https://proteo.ca/en/news-and-events/symposium-annuel-de-proteo-2023/#:~:text=Itiswithgreatpleasurethat | https://event.fourwaves.com/fr/proteo2023/resumes/3baffab4-6a26-4e21-b47a-1a3288e73469/affiche | The non-conventional yeast Yarrowia lipolytica is gaining interest in biotechnology as a workhorse to produce recombinant protein, lipids and other biomolecules. Site-specific genome editing is however limited in this yeast. This was much improved by the recent development of a CRISPR-Cas9 genome editing protocol based on a tRNA-sgRNA fusion and in which Cas9 is under the control of a synthetic hybrid promoter, pUAS1B8-TEF(136). Although reported to be strong, the tandem repeats of pUAS1B8-TEF(136) are prone to cause polymerase slippage, molecular rearrangements and cloning difficulties. Here we report another synthetic promoter to drive the expression of Cas9, pTEF(366)-Kozak, a 3’-truncated version of the pTEF(406) promoter of Y. lipolytica fused to a synthetic Kozak sequence. Our comparison of the promoters’ strength using hrGFP reporters and RT-qPCR revealed that both the synthetic pTEF(366)-Kozak and pTEF(406) are at least 5-folds higher expression than the synthetic pUAS1B8-TEF(136). Moreover, Cas9 under pTEF(366)-Kozak resulted in higher genome editing efficiencies by 40% compared to using pUAS1B8-TEF(136). Phenotypic analysis upon gene deletions revealed no significant difference between promoters as phenotypic losses were at their maxima. Here, we demonstrate that higher Cas9 expression is beneficial without showing any growth defect in enhancing CRISPR-Cas9 mediated genome editing in Y. lipolytica. This is the first study conducting rational 3’-truncation in TEF promoter based on in silico predictions of its structure, which can be extended for the engineering of other yeast promoters with the goal of generating small-sized synthetic biology parts for convenient engineering of biological systems. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-49688dcd451ce0f1b698567192ff56ce-ff-2023-05-04_CasExpression-Poster.pdf | no | |||||||
| 2023-05 | Lipid profiling of Yarrowia lipolytica mutants for the production of designer biodiesel | Oral Presentation | Invited | Synthetic Biology Club, Abdel-Mawgoud Synthetic Biology Lab, Université Laval, Québec, Canada | Abdel-Mawgoud SB Club | https://abdel-mawgoud.com/en/synthetic-biology-club/ | Biodiesels are a growing fuel class with the advancement of sustainable and renewable energy. Despite the advantages they hold over petroleum diesel, such as higher cetane number and reduced emissions, the current biodiesels are limited mainly in their cold flow properties. Advances in the 4 generations of biodiesel as well as the chemical and physical modification of feedstock oil in the production of designer biodiesel have however resulted in biodiesels of low stability or greater cost. With the promising approach of biodiesel production through genetic modification, here we genetically modified the oleaginous yeast Yarrowia lipolytica in order to shift its lipid composition towards biodiesel with improved properties. Mutations mfe1, pex10 and pox2 generated lipid profiles associated with improved flow properties despite a small reduction in cetane number predicted by mathematical regressions. Statistical analysis based on full-facorial regressions of biodiesel properties in function of average biodiesel composition led to the identification of potential optimal composition of 17.63 average chain length and 1.29 – 1.51 average unsaturation degree. This study shows the potential of genetically driven improvement of designer biodiesel in order to produce more efficient and reliable biofuel. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-8cb9507aeca302b68e66a55977206471-ff-OuelletB_SBClub-2023-main.png | no | ||||||
| 2023-08 | Optimizing microbial lipid for production of high-performance biodiesel in Yarrowia lipolytica | Oral Presentation | Journée étudiante de l’IBIS (2023), Université Laval, Québec, Canada. | https://journee.ibis.ulaval.ca/edition-2023-2/ | Biodiesels are a growing fuel class in a world increasingly shifting to ecological, sustainable, and renewable energy. Despite their advantages over petroleum diesel, such as higher cetane number and reduced emissions, current biodiesels are limited mainly in their cold flow properties, their larger emissions of NOx and poorer oxidative stability. Advances in the 4 generations of biodiesel as well as the chemical and physical modification of feedstock oil in the production of designer biodiesel mixtures have however resulted in biodiesels of ethical concerns, low stability and/or greater cost. With the promising approach of biodiesel production through genetic modification, here we genetically modified the oleaginous yeast Yarrowia lipolytica in order to shift its lipid composition towards biodiesel with improved properties, namely genetically modified designer biodiesel or simply Genetically Engineered Biodiesel (GEB). Mutation of the MFE1, PEX10 and POX2 genes generated lipid profiles with lower average chain length, and higher unsaturation degree associated with GEB of improved flow properties (7 °C lower than PO1f) despite a small reduction in cetane number (58 for PO1f compared to ~54 for mutants) as predicted by mathematical models. Statistical analysis based on full-factorial regressions of biodiesel properties as a function of average biodiesel composition led to the identification of potential optimal composition of biodiesel to be of an average carbon chain length of 17.4 and of an average unsaturation degree of 1.33. This study shows the potential of genetically driven improvement of designer biodiesel in order to produce more efficient and reliable biofuel. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-16010308e3947d36a94d3e6cc6225df7-ff-OuelletB_IBIS-2023-main.png | yes | Monetary | 220 | ||||||
| 2023-09 | Optimizing microbial lipid for production of high-performance biodiesel in Yarrowia lipolytica | Oral Presentation | Journée de la Recherche en Sciences et Génie (2023), Université Laval, Québec, Canada. | JRSG 2023 | Biodiesels are a growing fuel class in a world increasingly shifting to ecological, sustainable, and renewable energy. Despite their advantages over petroleum diesel, such as higher cetane number and reduced emissions, current biodiesels are limited mainly in their cold flow properties, their larger emissions of NOx and poorer oxidative stability. Advances in the 4 generations of biodiesel as well as the chemical and physical modification of feedstock oil in the production of designer biodiesel mixtures have however resulted in biodiesels of ethical concerns, low stability and/or greater cost. With the promising approach of biodiesel production through genetic modification, here we genetically modified the oleaginous yeast Yarrowia lipolytica in order to shift its lipid composition towards biodiesel with improved properties, namely genetically modified designer biodiesel or simply Genetically Engineered Biodiesel (GEB). Mutation of the MFE1, PEX10 and POX2 genes generated lipid profiles with lower average chain length, and higher unsaturation degree associated with GEB of improved flow properties (7 °C lower than PO1f) despite a small reduction in cetane number (58 for PO1f compared to ~54 for mutants) as predicted by mathematical models. Statistical analysis based on full-factorial regressions of biodiesel properties as a function of average biodiesel composition led to the identification of potential optimal composition of biodiesel to be of an average carbon chain length of 17.4 and of an average unsaturation degree of 1.33. This study shows the potential of genetically driven improvement of designer biodiesel in order to produce more efficient and reliable biofuel. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-93e141e241692167367a11c6c0057c5c-ff-OuelletB_JRSG-2023-main.png | no | ||||||||
| 2024-01 | Multiplexed CRISPR: Complex genome engineering and diverse applications | Oral Presentation | Invited | Synthetic Biology Club, Abdel-Mawgoud Synthetic Biology Lab, Université Laval, Québec, Canada | Abdel-Mawgoud SB Club | https://abdel-mawgoud.com/en/synthetic-biology-club/ | The development of CRISPR-based technology has revolutionized genome engineering, enabling efficient modification of various microorganisms with limited to no scars. As iterations of CRISPR-mediated genetic engineering can be time-consuming, Multiplexed CRISPR allows for targeting multiple genes at once by expressing different gene-specific gRNA. This technique has proven to be effective in multiplex gene disruption, gene integration, as well as CRISPR interference and activation. In this talk, we’ll overview the concept of Multiplexed CRISPR, strategies available and its applications. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-2e695f6a4c06f7da77d6908c00df8ac5-ff-OuelletB_SBClub-2024-main.png | no | ||||||
| 2024-05 | Production of genetically engineered designer biodiesel adapted for cold climates | Poster Presentation | Symposium Annuel de l’Institut nordique du Québec (2024), Museum of civilization, Quebec, QC, Canada | INQ annual symposium 2024 | https://inq.ulaval.ca/fr/symposium-inq-2024 | Biodiesels constitute a growing class of fuel in a world that is increasingly inclined towards more ecological and sustainable energies. Despite their advantages, biodiesels are especially limited in their operability in cold environment which is attributed to their chemical compositions stemming from their feedstock oils. This study provides a proof-of-concept for the use of genetic engineering to tailor biodiesel properties and performances in the development of standalone optimised energy for the North. This will enable the efficient and eco-responsible adaptation of energy related to transport in northern territory, while promoting local energy production. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-535bfa008f61a1d14208cfd1dd7c80ba-ff-2024-03-25_poster-biodiesel-AS.pdf | no | |||||||
| 2024-08 | Production of genetically engineered designer biodiesel adapted for cold climates | Poster Presentation | Journée étudiante de l’IBIS (2024), Université Laval, Québec, Canada. | https://journee.ibis.ulaval.ca/edition-2024 | Biodiesels constitute a growing class of fuel in a world that is increasingly inclined towards more ecological and sustainable energies. Despite their advantages, biodiesels are especially limited in their operability in cold environment which is attributed to their chemical compositions stemming from their feedstock oils. This study provides a proof-of-concept for the use of genetic engineering to tailor biodiesel properties and performances in the development of standalone optimised energy for the North. This will enable the efficient and eco-responsible adaptation of energy related to transport in northern territory, while promoting local energy production. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-87013d6ac644bf4aa26c32e4d46ee59a-ff-2024-03-25_poster-biodiesel-AS.pdf | no | ||||||||
| 2024-08 | Cold-induced modification of yeast lipids to produce optimized biodiesels | Oral Presentation | Journée étudiante de l’IBIS (2024), Université Laval, Québec, Canada. | https://journee.ibis.ulaval.ca/edition-2024 | Biodiesels constitute a growing class of fuel in a world that is increasingly inclined towards more ecological, sustainable, and renewable energies. Compared to diesel, biodiesels benefit from low ignition delay and cleaner combustion. However, those advantages come with high viscosity, large NOX emissions, low oxidation stability and, especially, limited cold flow properties. To overcome the latter drawback, current biodiesels are blended with petrodiesel which raises concerns of sustainability. The physicochemical properties of a given biodiesel stem from the chemical composition of its feedstock oil. Hence, we previously determined the optimal oil composition giving the best performing biodiesel; an oil mainly composed of unsaturated fatty acids with an average length of 17.36 carbons and an unsaturation degree of 1.33. In this study, we aimed to improve biodiesel properties by favoring unsaturated fatty acid production in Yarrowia lipolytica using control of cultivation conditions, namely incubation temperature. Analysis of the fatty acid content of Y. lipolytica strains at cold incubation temperatures revealed that, while the abondance of total unsaturated fatty acid content remained constant at 80% across all samples, we observed a two-fold increase in the proportion of polyunsaturated fatty acids reaching 60 %. Predicted properties of biodiesels produced under these cold cultivation conditions showed to meet international standards, enabling biodiesel operability at temperatures as low as -7°C without any blending with petrodiesel. This study shows a simple and efficient process for oil content modulation by cultivation under cold conditions for the production of biodiesels with optimised properties and performance. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-de6958db392321ca0303cd4602b9edc8-ff-OuelletB_IBIS-2024-main.png | no | ||||||||
| 2024-09 | Production of genetically engineered designer biodiesel adapted for cold climates | Poster Presentation | Journée de la Recherche en Sciences et Génie (2024), Université Laval, Québec, Canada. | JRSG 2024 | Biodiesels constitute a growing class of fuel in a world that is increasingly inclined towards more ecological and sustainable energies. Despite their advantages, biodiesels are especially limited in their operability in cold environment which is attributed to their chemical compositions stemming from their feedstock oils. This study provides a proof-of-concept for the use of genetic engineering to tailor biodiesel properties and performances in the development of standalone optimised energy for the North. This will enable the efficient and eco-responsible adaptation of energy related to transport in northern territory, while promoting local energy production. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-efa56ad7c9698991496e5fcaee65caff-ff-2024-03-25_poster-biodiesel-AS.pdf | no | ||||||||
| 2024-09 | Cold-induced modification of yeast lipids to produce optimized biodiesels | Oral Presentation | Journée de la Recherche en Sciences et Génie (2024), Université Laval, Québec, Canada. | JRSG 2024 | Biodiesels constitute a growing class of fuel in a world that is increasingly inclined towards more ecological, sustainable, and renewable energies. Compared to diesel, biodiesels benefit from low ignition delay and cleaner combustion. However, those advantages come with high viscosity, large NOX emissions, low oxidation stability and, especially, limited cold flow properties. To overcome the latter drawback, current biodiesels are blended with petrodiesel which raises concerns of sustainability. The physicochemical properties of a given biodiesel stem from the chemical composition of its feedstock oil. Hence, we previously determined the optimal oil composition giving the best performing biodiesel; an oil mainly composed of unsaturated fatty acids with an average length of 17.36 carbons and an unsaturation degree of 1.33. In this study, we aimed to improve biodiesel properties by favoring unsaturated fatty acid production in Yarrowia lipolytica using control of cultivation conditions, namely incubation temperature. Analysis of the fatty acid content of Y. lipolytica strains at cold incubation temperatures revealed that, while the abondance of total unsaturated fatty acid content remained constant at 80% across all samples, we observed a two-fold increase in the proportion of polyunsaturated fatty acids reaching 60 %. Predicted properties of biodiesels produced under these cold cultivation conditions showed to meet international standards, enabling biodiesel operability at temperatures as low as -7°C without any blending with petrodiesel. This study shows a simple and efficient process for oil content modulation by cultivation under cold conditions for the production of biodiesels with optimised properties and performance. | B. Ouellet*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-43b0e4c166755bf51d943b393b37635f-ff-OuelletB_JRSG-2024-main.png | no | ||||||||
| 2025-05 | Leveraging cold-induced changes in yeast lipid composition for the production of enhanced biodiesel variants. | Oral Presentation | PROTEO Annual Symposium (2025), University of Sherbrooke's Health Campus, QC, Canada | https://proteo.ca/en/news-and-events/24th-annual-proteo-symposium/ | https://event.fourwaves.com/fr/proteo2023/resumes/3baffab4-6a26-4e21-b47a-1a3288e73469/affiche | Abstract:Lipids are major components of cell membranes whose chemical structures are modulated with respect to their fatty acid chain length and unsaturation level to maintain, amongst others, membrane fluidity under variable temperatures. Such structural modifications can be leveraged to support the production of enhanced biodiesel variants whose performance and physicochemical properties are dictated by the structures of fatty acids from which biodiesels are made.This study aimed at developing a strategy in which the oleaginous yeast Yarrowia lipolytica is exposed to cold stress as a means to enhance the unsaturated fatty acid content in this yeast’s oils. Changes in oil profile were studied in this yeast when grown at 4 °C compared to that when grown at 28 °C. Fatty acid analysis revealed that cold-stress exposure resulted in an increased unsaturated-to-saturated fatty acid (UFA/SFA) ratio in all tested yeast strains and cultivation media (PO1f and mutant mfe1Δ strains; rich and lipogenic media). Overall, linoleates constituted more than 50 %wt of total fatty acids content by weight and minimal changes in overall chain length were observed. Though cold stress delayed growth, similar biomass concentrations were ultimately attained with improved lipid productivity. Biodiesels made from unsaturated-fatty-acid-rich lipids tend to perform better under cold operating conditions, since saturated-fatty acid-rich biodiesels tend to gel or crystallize at low temperatures.This study shows how a native lipid-profile adaptation, that a cell adopts to preserve its membrane fluidity under cold stress, can be leveraged for the production of enhanced oils for biodiesel suited for cold related properties. | B. Ouellet*; A. M. Abdel-Mawgoud | https://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-1e63e483f62f7d099238874ebc7b2800-ff-OUELLETBenjamin_PROTEO-symposium-2025.jpg | no | |||||||
| 2025-08 | Engineering diverse oils in Yarrowia lipolytica for tailored biodiesel production | Oral Presentation | Journée étudiante de l’IBIS (2025), Université Laval, Québec, Canada. | https://journee.ibis.ulaval.ca/edition-2025 | Fatty acids are valuable biomolecules in the agri-food, cosmetic, and energy sectors, notably as fatty acid methyl esters. The synthetic modification of fatty acids is of interest to produce long-chain polyunsaturated fatty acids, including eicosapentaenoic, arachidonic, and docosahexaenoic acid, for their nutritional value. Similarly, unsaturated fatty acids exhibit favorable properties for use as biodiesel owing to their viscosity and cold-flow characteristics.In this study, fatty acid-modifying genes were introduced in the oleaginous yeast Yarrowia lipolytica to diversify its lipid profile for biodiesel development. In-phase yeast oil extracts were analyzed both quantitatively and qualitatively using GC-FID/EI-MS. While fatty acid species associated with the synthetic Δ5Δ6 and Δ12Δ15 desaturase activity such as α-linolenic and γ-linolenic acids were detected, their concentrations remained in trace amounts, representing less than 0.1% of the total fatty acid content in the PO1f strain. Disruption of the β-oxidation pathway via the mfe1Δ mutation led to a modest increase in their levels, though still below 0.5%. Aside from the fatty acids introduced through genetic engineering, only minor changes were observed in the overall oil composition and the resulting biodiesel properties, with no significant impact on yield reaching 10% and 25% of dry cell mass in the PO1f and mfe1Δ variants, respectively.These findings suggest that genetic engineering using fatty acid-modifying genes has limited effectiveness in diversifying oil composition for biodiesel production. In contrast, simpler strategies, such as adjusting environmental parameters like temperature and oxygenation, may offer more effective and flexible means of tailoring lipid profiles for customized biodiesel applications. | B. Ouellet*; A. M. Abdel-Mawgoud | https://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-f0920c0dcd3b03c0409edfa5bb280f93-ff-OUELLETBenjamin_JRIBIS-2025.jpg | no | ||||||||
| 2025-09 | Engineering diverse oils in Yarrowia lipolytica for tailored biodiesel production | Oral Presentation | Journée de la Recherche en Sciences et Génie (2025), Université Laval, Québec, Canada. | JRSG 2025 | https://aegseg.asso.ulaval.ca/jrsg/jrsg-2025/ | Fatty acids are valuable biomolecules in agri-food, cosmetic, and energy industries, notably in the form of fatty acid methyl esters. Modification of fatty acids to produce long-chain polyunsaturated fatty acids, such as arachidonic acid, is of interest for their nutritional value. Similarly, unsaturated fatty acids exhibit favorable properties for use as cold-climate-adapted biodiesel owing to their lower viscosity and better cold-flow properties.In this study, heterologous fatty acid-modifying genes were introduced in the oleaginous yeast Yarrowia lipolytica to diversify its lipid profile for biodiesel improvement. While fatty acid species associated with the heterologous Δ5Δ6 and Δ12Δ15 desaturase activity such as α-linolenic and γ-linolenic acids were detected, their concentrations remained in trace amounts in the PO1f strain. In mfe1Δ mutants, disruption of the β-oxidation pathway led to a modest increase reaching around 1% of the total fatty acid content. Besides, only minor changes were observed in the overall oil composition and the resulting biodiesel properties, with no significant impact on yield reaching 10% and 25% of dry cell mass in the PO1f and mfe1Δ variants, respectively.These findings suggest that genetic engineering of lipid profiles might be of limited significance when targeting a limited number of fatty acid-modifying genes for biodiesel production. In contrast, engineering of multiple genes could result in more significant effects on lipid profiles. Alternatively, simpler strategies, such as adjusting environmental parameters like temperature and oxygenation, might offer more effective and convenient means of tailoring lipid profiles for customized biodiesel applications. | B. Ouellet*; A. M. Abdel-Mawgoud | https://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-f5f91d7ae7a088debb1f845706ce4554-ff-OUELLETBenjamin_JRSG-2025.jpg | no | |||||||
| 2023-06 | Medium optimization for high lipid production by Rhodotorula toruloides and Yarrowia lipolytica using an optimized Nile Red-based lipid fluorometry | Poster Presentation | Annual Synthetic Biology Symposium (2023), Loyola campus, Concordia University, Montréal, Canada | SynBio 5.0 | https://sites.events.concordia.ca/sites/synbio/en/synbio5-0/home#:~:text=SynBioWeekcombinesSynbio5.0in | As lipogenic yeasts are becoming increasingly harnessed as biofactories of oleochemicals, the availability of efficient protocols for the determination and optimization of lipid titers in these organisms is necessary. In this study, we optimized a quick, reliable and high throughput Nile Red-based lipid fluorometry protocol adapted for oleaginous yeasts and validated it using different approaches, the most important of which is using gas chromatography coupled to flame ionization detection. This protocol was applied for the optimization of the concentrations of ammonium chloride and glycerol for the highest lipid titers in Rhodotorula toruloides NRRL Y-6987 and Yarrowia Lipolytica W29 using response surface central composite design (CCD). The optimal concentration of ammonium chloride and glycerol are 4 and 123 g/L achieving a C/N ratio of 57 for R. toruloides, whereas concentrations are 4 and 139 g/L with a C/N ration of 61 for Y. lipolytica. Outside the C/N of 33 to 74 and 45 to 75, respectively for R. toruloides and Y. lipolytica, lipid productions decrease by more than 10%. The developed regression models and response surface plots give an insight on the careful selection of optimal C/N ratio for maximal lipid production. | B. Ouellet*; Z. Morneau; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-575292bfd8d4f4d33906c10e9b40ce36-ff-2023-05-16_NileRed-AS.pdf | no | |||||||
| 2021-10 | Engineering of selenoneine in microbial hosts using synthetic biology. | Oral Presentation | Invited | Synthetic Biology Club, Abdel-Mawgoud Synthetic Biology Lab, Université Laval, Québec, Canada | Abdel-Mawgoud SB Club | https://abdel-mawgoud.com/en/ | Selenoneine, is an organic selenium compound, commonly found in the blood and tissues of several marine species. This molecule shows antioxidant activity against reactive oxygen species (ROS) and can neutralize mercury toxicity and is thus known for its health protective effects in many human diseases. Nonetheless, no economically viable means for selenoneine production has yet been established. The aim of this project is to develop synthetic biology strategies to engineer selenoneine-overproducing microbial hosts to allow its subsequent purification and characterization. To achieve this, design-build-test cycles for metabolic engineering will be iteratively repeated until desired selenoneine titers are produced, and the health protective effects of engineered strains will be investigated in zebra fish. This project will result in the development of engineered strains that might support a cheap, renewable, and scalable production of selenoneine. | T. García-Ybarra* ; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-5159fd289d15d9bf1b49032c630569dd-ff-2024-10-25.png | no | ||||||
| 2021-11 | Engineering of selenoneine in microbial hosts using synthetic biology | Oral Presentation | Invited | Journée de la Recherche en Sciences et Génie (2021), Université Laval, Québec, Canada. | JRSG 2021 | Selenoneine, is an organic selenium compound, commonly found in the blood and tissues of several marine species. This molecule shows antioxidant activity against reactive oxygen species (ROS) and can neutralize mercury toxicity and is thus known for its health protective effects in many human diseases. Nonetheless, no economically viable means for selenoneine production has yet been established. The aim of this project is to develop synthetic biology strategies to engineer selenoneine-overproducing microbial hosts to allow its subsequent purification and characterization. To achieve this, design-build-test cycles for metabolic engineering will be iteratively repeated until desired selenoneine titers are produced, and the health protective effects of engineered strains will be investigated in zebra fish. This project will result in the development of engineered strains that might support a cheap, renewable, and scalable production of selenoneine. | T. García-Ybarra* ; A. M. Abdel-Mawgoud | no | ||||||||
| 2021-11 | Engineering of selenoneine in microbial hosts using synthetic biology | Oral Presentation | Invited | Recruitment Day (2021), Department of Biochemistry, Microbiology and Bioinformatics, Faculty of Science and Engineering, Université Laval, Québec, Canada. | Selenoneine, is an organic selenium compound, commonly found in the blood and tissues of several marine species. This molecule shows antioxidant activity against reactive oxygen species (ROS) and can neutralize mercury toxicity and is thus known for its health protective effects in many human diseases. Nonetheless, no economically viable means for selenoneine production has yet been established. The aim of this project is to develop synthetic biology strategies to engineer selenoneine-overproducing microbial hosts to allow its subsequent purification and characterization. To achieve this, design-build-test cycles for metabolic engineering will be iteratively repeated until desired selenoneine titers are produced, and the health protective effects of engineered strains will be investigated in zebra fish. This project will result in the development of engineered strains that might support a cheap, renewable, and scalable production of selenoneine. | T. García-Ybarra* ; A. M. Abdel-Mawgoud | no | |||||||||
| 2021-11 | Engineering of selenoneine in microbial hosts using synthetic biology. | Oral Presentation | Invited | Réunion scientifique Sentinelle Nord 2021, Online, Québec, Canada | https://sentinelnorth.ulaval.ca/en/scientific-meeting-2021 | Selenoneine, is an organic selenium compound, commonly found in the blood and tissues of several marine species. This molecule shows antioxidant activity against reactive oxygen species (ROS) and can neutralize mercury toxicity and is thus known for its health protective effects in many human diseases. Nonetheless, no economically viable means for selenoneine production has yet been established. The aim of this project is to develop synthetic biology strategies to engineer selenoneine-overproducing microbial hosts to allow its subsequent purification and characterization. To achieve this, design-build-test cycles for metabolic engineering will be iteratively repeated until desired selenoneine titers are produced, and the health protective effects of engineered strains will be investigated in zebra fish. This project will result in the development of engineered strains that might support a cheap, renewable, and scalable production of selenoneine. | T. García-Ybarra* ; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-8e5f1aec78ecb1770ed2653985640a11-ff-2024-11.png | no | |||||||
| 2022-11 | Engineering of selenoneine in microbial hosts using synthetic biology | Oral Presentation | Invited | Recruitment Day (2022), Department of Biochemistry, Microbiology and Bioinformatics, Faculty of Science and Engineering, Université Laval, Québec, Canada. | Selenoneine, is an organic selenium compound, commonly found in the blood and tissues of several marine species. This molecule shows antioxidant activity against reactive oxygen species (ROS) and can neutralize mercury toxicity and is thus known for its health protective effects in many human diseases. Nonetheless, no economically viable means for selenoneine production has yet been established. The aim of this project is to develop synthetic biology strategies to engineer selenoneine-overproducing microbial hosts to allow its subsequent purification and characterization. To achieve this, design-build-test cycles for metabolic engineering will be iteratively repeated until desired selenoneine titers are produced, and the health protective effects of engineered strains will be investigated in zebra fish. This project will result in the development of engineered strains that might support a cheap, renewable, and scalable production of selenoneine. | T. García-Ybarra* ; A. M. Abdel-Mawgoud | no | |||||||||
| 2023-08 | A GC-MS/FID method for the determination of essential and antioxidant thio- and seleno-amino acids in biological samples | Poster Presentation | Journée étudiante de l’IBIS (2023), Faculty of Science and Engineering, Université Laval, Québec, Canada. | https://journee.ibis.ulaval.ca/edition-2023-2/ | Essential amino acids are the building blocks of proteins and are present in diverse biological matrices. From essential amino acids derive other essential amino acids that have interesting biological activities for research. For example, ergothioneine (EGT) and selenoneine (SEN) are thio- and seleno-amino acids derived from histidine and are known to have interesting antioxidant properties valuable for medicinal and commercial applications. The methods available for the analysis of antioxidant thio- and seleno-amino acids are mostly based on costly LC-MS method which is slowing down advances in research in such important amino acids. In general, GC methods are less expensive, more resolving, and more accessible for the accurate analysis of amino acids compared to LC-based methods. Although several GC methods are available for the analysis of essential amino acids, yet no universal GC-based method is available for the analysis of essential amino acids together with EGT and SEN and their metabolic precursors. Even though GC methods require prior derivatization of amino acids to enhance their volatility in GC columns, yet LC-MS methods are also increasingly relying on prior derivatization of amino acid to increase their chromatographic resolution.In this study, we developed a rapid and less expensive GC-MS method, using full ion scan or SIM, for the sensitive detection and precise quantification of all essential as well as certain antioxidant thio- and seleno-amino acids in biological samples in a single run. Moreover, we adapted the method for the far less expensive GC-FID instruments. The method involves the development of an efficient derivatization step using isobutyl chloroformate (IBCF) as the alkylating agent. We further validated our method using the LC-HRMS using orbitrap detector that demonstrated the completion of derivatization of amino acids to 100% and helped confirming the identity of SEN amino acids for which no standards are commercially available. The method was used for the identification of several microbial producers of antioxidant amino acids.Due to its rapidity and low cost, our method increases the range of metabolites that can be analyzed in GC-MS-based metabolomic analyses. Moreover, our method is believed to boost discovery, metabolic engineering, and synthetic biology research on antioxidant amino acids. | T. García-Ybarra* ; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-7217234d8e0a9af61c29e8c55568c187-ff-2023-08.png | yes | Monetary | 200 | ||||||
| 2023-09 | Engineering of selenoneine in microbial hosts using synthetic biology | Poster Presentation | Journée de la Recherche en Sciences et Génie (2023), Université Laval, Québec, Canada. | JRSG 2023 | Selenoneine, is an organic selenium compound, commonly found in the blood and tissues of several marine species. This molecule shows antioxidant activity against reactive oxygen species (ROS) and can neutralize mercury toxicity and is thus known for its health protective effects in many human diseases. Nonetheless, no economically viable means for selenoneine production has yet been established. The aim of this project is to develop synthetic biology strategies to engineer selenoneine-overproducing microbial hosts to allow its subsequent purification and characterization. To achieve this, design-build-test cycles for metabolic engineering will be iteratively repeated until desired selenoneine titers are produced, and the health protective effects of engineered strains will be investigated in zebra fish. This project will result in the development of engineered strains that might support a cheap, renewable, and scalable production of selenoneine. | T. García-Ybarra* ; A. M. Abdel-Mawgoud | no | |||||||||
| 2023-11 | Determination of antioxidant thio- and seleno-amino acids in selected strains | Oral Presentation | Invited | Réunion scientifique Sentinelle Nord (2023), Québec city Convention Center, Québec, Canada. | https://sentinelnorth.ulaval.ca/en/2023-scientific-meeting | Ergothioneine (EGT) and selenoneine (SEN) histidine-derived thio- and seleno-amino acids that demonstrate potent antioxidant and cytoprotective properties that find applications in food, medicinal and pharmaceutical applications. Currently, research efforts are being exerted to boost yield and purity of produced SEN and EGT for augmenting their commercial availability. Many native microorganisms are known to produce either EGT and/or SEN. The biosynthesis of EGT in bacteria requires the implication of five genes egtABCDE, whereas SEN, shares egtD.One of the important limitations slowing down research in SEN/EGT engineering is the lack of a cheap, precise method for the determination of these peculiar amino acids. The methods available for the analysis of antioxidant thio- and seleno-amino acids are mostly based on costly LC-MS method which is slowing down advances in research in such important amino acids. In general, GC methods are less expensive, more resolving, and more accessible for the accurate analysis of amino acids compared to LC-based methods. Although several GC methods are available for the analysis of essential amino acids, yet no universal GC-based method is available for the analysis of essential amino acids together with EGT and SEN and their metabolic precursors.In this study, we developed the first GC-MS method, using selected ion monitoring (SIM), for the simultaneous analysis of essential and antioxidant amino acids in a single run. In addition, we adapted the method for the far less expensive GC-FID instruments. Moreover, we applied and validated our method in the detection of EGT/SEN in selected strains.We believe our analytical method is an effective, convenient, and least expensive method to date for the determination of EGT and SEN in microbial strains. | T. García-Ybarra* ; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-8c70bc0a988541d5854846a94014020e-ff-Sentinel-North-2023.jpg | no | |||||||
| 2023-09 | Determination of antioxidant thio- and seleno-amino acids in selected bacteria | Poster Presentation | Congrès de Bactériologie intégrative: Symbiose – Pathogenèse, Pavillon Roger Gaudry, Université de Montréal, Québec, Canada. | Congrès BiSP 2023 | https://event.fourwaves.com/fr/bisp2023/pages | From essential amino acids derive special antioxidant amino acids have interesting biological activities. Namely, ergothioneine (EGT) and selenoneine (SEN) histidine-derived thio- and seleno-amino acids that demonstrate potent antioxidant and cytoprotective properties that find applications in food, medicinal and pharmaceutical applications. Currently, research efforts are being exerted to boost yield and purity of produced SEN and EGT for augmenting their commercial availability. Many native microorganisms are known to produce either EGT and/or SEN. The biosynthesis of EGT in bacteria requires the implication of five genes egtABCDE, whereas SEN, shares egtD. In bacteria, it is now clear that EgtD trimethylates the NH2 group of histidine to generate hercynine. Then EgtA catalyzes the synthesis of hercynylcysteine sulfoxide, which in the presence of pyridoxal 5-phosphate, is converted into EGT by EgtE. By contrast, the biosynthetic pathway for SEN, EgtD trimethylates histidine to hercyncyl- selenoxide which can spontaneously be converted to SEN or be reduced to hercynyl- selenoether. One of the important limitations slowing down research in SEN/EGT engineering is the lack of a cheap, precise method for the determination of these peculiar amino acids. The methods available for the analysis of antioxidant thio- and seleno-amino acids are mostly based on costly LC-MS method which is slowing down advances in research in such important amino acids. In general, GC methods are less expensive, more resolving, and more accessible for the accurate analysis of amino acids compared to LC-based methods. Although several GC methods are available for the analysis of essential amino acids, yet no universal GC-based method is available for the analysis of essential amino acids together with EGT and SEN and their metabolic precursors.In this study, we developed the first GC-MS method, using selected ion monitoring (SIM), for the simultaneous analysis of essential and antioxidant amino acids in a single run. In addition, we adapted the method for the far less expensive GC-FID instruments. Moreover, we applied and validated our method in the detection of EGT/SEN in selected bacterial strains.We believe our analytical method is an effective, convenient, and least expensive method to date for the determination of EGT and SEN in microbial strains. This method is hence expected to accelerate research on engineering of SEN and EGT aiming at bringing these antioxidants into market for commercial use. | T. García-Ybarra*; A. M. Abdel-Mawgoud | http://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-2605470252536f11ab55b2032005e5a1-ff-2023-08.png | no | |||||||
| 2024-06 | An approach to the production of genetically engineered biodiesel in lipogenic yeasts | Oral Presentation | Invited | MicroBioMoléculaires meetings organized by microbiology and molecular biology research groups of Biology department at University of Sherbrooke, QC Canada. | MicroBioMoléculaires meetings | Biodiesels are increasingly considered as more sustainable and ecological alternatives to fossil fuels. Although biodiesels have many advantages over petrodiesel, such as higher cetane number and 75% less CO2 emissions, they have higher NOX emissions, and they can not be used as standalone fuels mainly because of their limited cold flow properties. These limitations are directly attributed to the chemical compositions of biodiesels that stems from that of their feedstock oils. Modification of chemical composition of feedstock oils at their biological sources can thus enable the production of designer biodiesel with improved physicochemical properties, performance, and emission metrics. Using mathematical models and full-factorial central composite design, the optimal biodiesel feedstock oil composition was predicted. This optimum composition was respectfully targeted by engineering oil composition of the oleaginous yeast, Yarrowia lipolytica. In this yeast, simple and combined mutations of certain lipid-associated genes generated fatty acid profiles whose biodiesel has better predicted cold flow properties despite a small reduction in cetane number that is still within standard limits. Moreover, several mutants showed 2–3 fold higher lipid titers compared to the parent strain. This study is laying a foundational approach for the production of genetically engineered biodiesel (GEB) with desired properties and performance metrics, which ultimately finds solutions enabling the use of biodiesel as a standalone fuel. | A. M. Abdel-Mawgoud | https://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-3fbd64ce5b275d7827c5e6f1352e30fa-ff-2024-06-11-Production-of-designer-biodiesel-Ahmad-Saleh.jpg | no | |||||||
| 2025-06 | AMSBL platform for chemical analysis: GC-MS analysis | Oral Presentation | Invited | Retreat Day of the Institut de biologie integrative et de système, IBIS, Complex Desjardins, Laval University, Québec, QC, Canada | This talk provides an overview of Gas Chromatography-Mass Spectrometry (GC-MS) method of analysis as part of the services offered by AMSBL analytical platform. GC-MS is a powerful technique widely utilized for the identification and quantification of volatile and semi-volatile compounds in complex mixtures. We will introduce the fundamental principles of GC-MS , explaining the roles of the gas chromatograph in separating components based on their physiochemical properties and the mass spectrometer in generating characteristic fragmentation patterns (mass spectra) for compound identification. Key parameters affecting separation and sensitivity will be discussed. The talk will highlight the diverse applications of GC-MS across various fields, including environmental analysis (e.g., pollutant detection), forensic science (e.g., drug identification), quality control in food and fragrance industries (e.g., flavor/aroma profiling), and particularly in chemical and materials science research for characterizing organic synthesis products and complex material extracts. We will provide examples of relevant research areas where GC-MS provides indispensable data. Finally, we will detail the specific GC-MS analytical services available through the AMSBL platform. This includes sample preparation guidelines, instrument specifications, methods for both qualitative identification and quantitative analysis, and how researchers can access and utilize the platform's expertise and resources to advance their analytical objectives. | A. M. Abdel-Mawgoud | https://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-17c91f23884ce981f6fb166bb1af2994-ff-Frontpage.jpg | no | ||||||||
| 2025-12 | Metabolic Engineering of Yeasts for Biodiesel and Rhamnolipid Production. | Oral Presentation | Invited | Discussion Group Seminar Series of Department of Biochemistry and Molecular Biology, Faculty of Medicine, and Michael Smith Laboratories, Michael Smith Laboratories Auditorium, University of BC, Vancouver, BC, Canada | BMBDG-MSL Seminar Series | https://biochem.ubc.ca/2025-26-seminars/ | https://biochem.ubc.ca/bmb-msl-seminar-dr-ahmad-abdel-mawgoud/ | Biodiesel (Fatty Acid Methyl Esters) is a renewable, cleaner-burning alternative to petroleum diesel, but its inherent poor cold-flow properties—driven by the composition of conventional plant-based feedstock oils—currently prevent its use as a standalone fuel in cold climates. We addressed this limitation by engineering oleaginous yeast oil, which offers significant ethical and technical advantages over modifying plant lipids. We first utilized mathematical models and full-factorial central composite design to predict the optimal cold-adapted fatty acid composition required for enhanced fuel performance. Subsequently, we engineered Yarrowia lipolytica by introducing simple and combined mutations in target lipid-associated genes to achieve this predicted profile. This metabolic engineering successfully generated modified fatty acid profiles whose resultant biodiesel is predicted to have enhanced cold-flow properties while maintaining a small, standard-compliant reduction in cetane number. Notably, several constructed mutants exhibited a remarkable 2–3 fold increase in total lipid titers compared to the parent strain. Ultimately, this research establishes a foundational methodology for producing genetically engineered biodiesel (GEB) with the necessary functional and economic metrics to achieve its widespread adoption as a standalone fuel. We will also present an overview of another ongoing project focused on the Metabolic Engineering of Yeasts for Biodiesel and Rhamnolipid Production | A. M. Abdel-Mawgoud | https://abdel-mawgoud.com/wp-content/uploads/fluentform/ff-8b8712919e79b2e0e334046d3a3d946a-ff-Front-slide.jpg | no |