Production of triacylglycerols in Escherichia coli by deletion of the diacylglycerol kinase gene and heterologous overexpression of atfA from Acinetobacter baylyi ADP1

Janßen, Helge Jans; Steinbüchel, Alexander

doi:10.1007/s00253-013-5460-2

Cited by 24 publications

(14 citation statements)

References 37 publications

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“…coli strains with deleted genes [17–20] and in wild-type (WT) E . coli strains overexpressing additional proteins [12,19,21,22]. However, recombinant oil production through the heterologous expression of a WS/DGAT has not been achieved in WT E .…”

Section: Introductionmentioning

confidence: 99%

Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli

et al. 2017

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Triglycerides (TAGs), the major storage molecules of metabolic energy and source of fatty acids, are produced as single cell oil by some oleogenic microorganisms. However, these microorganisms require strict culture conditions, show low carbon source flexibilities, lack efficient genetic modification tools and in some cases pose safety concerns. TAGs have essential applications such as behaving as a source for added-value fatty acids or giving rise to the production of biodiesel. Hence, new alternative methods are urgently required for obtaining these oils. In this work we describe TAG accumulation in the industrially appropriate microorganism Escherichia coli expressing the heterologous enzyme tDGAT, a wax ester synthase/triacylglycerol:acylCoA acyltranferase (WS/DGAT). With this purpose, we introduce a codon-optimized gene from the thermophilic actinomycete Thermomonospora curvata coding for a WS/DGAT into different E. coli strains, describe the metabolic effects associated to the expression of this protein and evaluate neutral lipid accumulation. We observe a direct relation between the expression of this WS/DGAT and TAG production within a wide range of culture conditions. More than 30% TAGs were detected within the bacterial neutral lipids in 90 minutes after induction. TAGs were observed to be associated with the hydrophobic enzyme while forming round intracytoplasmic bodies, which could represent a bottleneck for lipid accumulation in E. coli. We detected an increase of almost 3-fold in the monounsaturated fatty acids (MUFA) occurring in the recombinant strains. These MUFA were predominant in the accumulated TAGs achieving 46% of the TAG fatty acids. These results set the basis for further research on the achievement of a suitable method towards the sustainable production of these neutral lipids.

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Section: Introductionmentioning

confidence: 99%

Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli

et al. 2017

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“…This strategy has been applied in the cyanobacterium Synechocystis as well, thereby confirming its broad applicability (Liu et al, 2011). In addition, heterologous expression of the wax ester synthase atfA may make it possible to produce fatty acid esters in some methanotrophic strains, as well as triacylglycerides if a diacylglycerol pool is present or generated (Janssen and Steinbuchel, 2014;Steen et al, 2010). Intriguingly, because methanol is generated from methane in methanotrophs, 513 it may be possible to produce fatty acid methyl esters (FAMEs) in situ without additional 514 modifications for alcohol synthesis (Fassel et al, 1992).…”

Section: Production Of Fatty Acid-derived Compounds From Methanotrophsmentioning

confidence: 70%

Metabolic engineering in methanotrophic bacteria

Kalyuzhnaya

Puri

Lidstrom

2015

Metabolic Engineering

273

162

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Methane, as natural gas or biogas, is the least expensive source of carbon for (bio)chemical synthesis. Scalable biological upgrading of this simple alkane to chemicals and fuels can bring new sustainable solutions to a number of industries with large environmental footprints, such as natural gas/petroleum production, landfills, wastewater treatment, and livestock. Microbial biocatalysis with methane as a feedstock has been pursued off and on for almost a half century, with little enduring success. Today, biological engineering and systems biology provide new opportunities for metabolic system modulation and give new optimism to the concept of a methane-based bio-industry. Here we present an overview of the most recent advances pertaining to metabolic engineering of microbial methane utilization. Some ideas concerning metabolic improvements for production of acetyl-CoA and pyruvate, two main precursors for bioconversion, are presented. We also discuss main gaps in the current knowledge of aerobic methane utilization, which must be solved in order to release the full potential of methane-based biosystems.

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“…It is also known that atfA can catalyze the synthesis of triacylglycerides (TAGs) from a diacylglyceride and an acyl‐CoA molecule (Stoveken et al, ) and recently has been shown to lead to an overproduction of TAGs in E. coli (Janßen and Steinbüchel, ). This adds additional complications for optimizing FAEEs production and implies that yield of FAEEs can be diminished by side reactions of the acyltransferase.…”

Section: Resultsmentioning

confidence: 99%

Enhancing fatty acid ethyl ester production in Saccharomyces cerevisiae through metabolic engineering and medium optimization

Thompson

Trinh

2014

Biotech & Bioengineering

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Biodiesels in the form of fatty acyl ethyl esters (FAEEs) are a promising next generation biofuel due to their chemical properties and compatibility with existing infrastructure. It has recently been shown that expression of a bacterial acyl-transferase in the established industrial workhorse Saccharomyces cerevisiae can lead to production of FAEEs by condensation of fatty acyl-CoAs and ethanol. In contrast to recent strategies to produce FAEEs in S. cerevisiae through manipulation of de novo fatty acid biosynthesis or a series of arduous genetic manipulations, we introduced a novel genetic background, which is comparable in titer to previous reports with a fraction of the genetic disruption by aiming at increasing the fatty acyl-CoA pools. In addition, we combined metabolic engineering with modification of culture conditions to produce a maximum titer of over 25 mg/L FAEEs, a 40% improvement over previous reports and a 17-fold improvement over our initial characterizations. Biotechnol. Bioeng. 2014;111: 2200-2208. © 2014 Wiley Periodicals, Inc.

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Production of triacylglycerols in Escherichia coli by deletion of the diacylglycerol kinase gene and heterologous overexpression of atfA from Acinetobacter baylyi ADP1

Cited by 24 publications

References 37 publications

Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli

Heterologous expression of a thermophilic diacylglycerol acyltransferase triggers triglyceride accumulation in Escherichia coli

Metabolic engineering in methanotrophic bacteria

Enhancing fatty acid ethyl ester production in Saccharomyces cerevisiae through metabolic engineering and medium optimization

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