Metabolic Engineering of Moss for Sustainable Oleic Acid-Rich Oil Production

Additional Authors

Jyoti Ranjan Behera, Department of Biological Sciences, East Tennessee State University, Johnson City, TN.

Abstract

Triacylglycerols (TAGs), the predominant storage lipids in plants, are integral to vegetable oil composition. The anticipated surge in vegetable oil demand by 2030 necessitates sustainable production strategies. Oleic acid, a vital omega-9 monounsaturated fatty acid with diverse industrial applications, faces challenges in current production methods due to its low prevalence in plant seed-based oils. This study focuses on metabolically engineering Physcomitrium patens (moss) to improve oil production, mostly TAG with a higher proportion of oleic acid through stable expression of these avocado-derived transcription factors and enzymes. We successfully demonstrated a significant increase in TAG content, particularly oleic acid, through transient co-expression of four avocado (Persea americana) genes, namely WRINKLED1 (WRI1), WRI2, diacylglycerol acyltransferases (DGAT1), and phospholipid: DGAT (PDAT1). We hypothesize that expressing avocado genes in moss will enhance oleic acid production, providing a sustainable alternative for the oleochemical industry. The cDNA of avocado genes will be cloned into an expression vector via gateway strategy followed by PEG-mediated protoplast transformation and screening. Positive cell lines will be cultured, and lipid analysis via thin-layer chromatography (TLC) followed by gas chromatography will be carried out for TAG quantification and fatty acid composition analysis. Lipid droplets will be visualized and quantified by Nile Red staining followed by confocal fluorescence microscopy. We expect to demonstrate a substantial increase in oleic acid production in mosses. The transgenic moss will provide an eco-friendly solution for the oleochemical industry, addressing the growing demand for oleic acid in an environmentally conscious manner.

Start Time

16-4-2025 9:00 AM

End Time

16-4-2025 11:30 AM

Presentation Type

Poster

Presentation Category

Science, Technology and Engineering

Student Type

Graduate Student - Masters

Faculty Mentor

Aruna Kilaru

Faculty Department

Biological Sciences

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Apr 16th, 9:00 AM Apr 16th, 11:30 AM

Metabolic Engineering of Moss for Sustainable Oleic Acid-Rich Oil Production

Triacylglycerols (TAGs), the predominant storage lipids in plants, are integral to vegetable oil composition. The anticipated surge in vegetable oil demand by 2030 necessitates sustainable production strategies. Oleic acid, a vital omega-9 monounsaturated fatty acid with diverse industrial applications, faces challenges in current production methods due to its low prevalence in plant seed-based oils. This study focuses on metabolically engineering Physcomitrium patens (moss) to improve oil production, mostly TAG with a higher proportion of oleic acid through stable expression of these avocado-derived transcription factors and enzymes. We successfully demonstrated a significant increase in TAG content, particularly oleic acid, through transient co-expression of four avocado (Persea americana) genes, namely WRINKLED1 (WRI1), WRI2, diacylglycerol acyltransferases (DGAT1), and phospholipid: DGAT (PDAT1). We hypothesize that expressing avocado genes in moss will enhance oleic acid production, providing a sustainable alternative for the oleochemical industry. The cDNA of avocado genes will be cloned into an expression vector via gateway strategy followed by PEG-mediated protoplast transformation and screening. Positive cell lines will be cultured, and lipid analysis via thin-layer chromatography (TLC) followed by gas chromatography will be carried out for TAG quantification and fatty acid composition analysis. Lipid droplets will be visualized and quantified by Nile Red staining followed by confocal fluorescence microscopy. We expect to demonstrate a substantial increase in oleic acid production in mosses. The transgenic moss will provide an eco-friendly solution for the oleochemical industry, addressing the growing demand for oleic acid in an environmentally conscious manner.