A metabolic engineering strategy for producing free fatty acids by theYarrowia lipolyticayeast based on impairment of glycerol metabolism

2017 ◽  
Vol 115 (2) ◽  
pp. 433-443 ◽  
Author(s):  
Evgeniya Y. Yuzbasheva ◽  
Elizaveta B. Mostova ◽  
Natalia I. Andreeva ◽  
Tigran V. Yuzbashev ◽  
Alexander S. Fedorov ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Metabolic Engineering ◽  
Free Fatty Acids ◽  
Glycerol Metabolism ◽  
Metabolic Engineering Strategy ◽  
Engineering Strategy

Construction of yeast producing patchoulol by global metabolic engineering strategy

2020 ◽  
Vol 117 (5) ◽  
pp. 1348-1356 ◽  
Author(s):  
Ryosuke Mitsui ◽  
Riru Nishikawa ◽  
Ryosuke Yamada ◽  
Takuya Matsumoto ◽  
Hiroyasu Ogino
Keyword(s):  
Metabolic Engineering ◽  
Metabolic Engineering Strategy ◽  
Engineering Strategy

scholarly journals Metabolic engineering strategies to produce medium-chain oleochemicals via acyl-ACP:CoA transacylase activity

2021 ◽  
Author(s):  
Qiang Yan ◽  
William Cordell ◽  
Michael Jindra ◽  
Dylan Courtney ◽  
Madeline Kuckuk ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Metabolic Engineering ◽  
Acyl Chain ◽  
Fatty Alcohols ◽  
Methyl Ketones ◽  
Beta Oxidation ◽  
Medium Chain ◽  
Genetic Modifications ◽  
Engineering Strategy ◽  
Transacylase Activity

Abstract Microbial lipid metabolism is an attractive route for producing aliphatic chemicals, commonly referred to as oleochemicals. The predominant metabolic engineering strategy centers on heterologous thioesterases capable of producing fatty acids of desired size. To convert acids to desired oleochemicals (e.g. fatty alcohols, ketones), metabolic engineers modify cells to block beta-oxidation, reactivate fatty acids as coenzyme-A thioesters, and redirect flux towards termination enzymes with broad substrate utilization ability. These genetic modifications narrow the substrate pool available for the termination enzyme but cost one ATP per reactivation - an expense that could be saved if the acyl-chain was directly transferred from ACP- to CoA-thioester. In this work, we demonstrate an alternative acyl-transferase strategy for producing medium-chain oleochemicals. Through bioprospecting, mutagenesis, and metabolic engineering, we developed strains of Escherichia coli capable of producing over 1 g/L of medium-chain free fatty acids, fatty alcohols, and methyl ketones using the transacylase strategy.

scholarly journals A metabolic engineering strategy for producing conjugated linoleic acids using the oleaginous yeast Yarrowia lipolytica

2017 ◽  
Vol 101 (11) ◽  
pp. 4605-4616 ◽  
Author(s):  
Nabila Imatoukene ◽  
Jonathan Verbeke ◽  
Athanasios Beopoulos ◽  
Abdelghani Idrissi Taghki ◽  
Brigitte Thomasset ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Yarrowia Lipolytica ◽  
Oleaginous Yeast ◽  
Conjugated Linoleic Acids ◽  
Metabolic Engineering Strategy ◽  
Engineering Strategy ◽  
Yeast Yarrowia Lipolytica

scholarly journals Chloroplastic metabolic engineering coupled with isoprenoid pool enhancement for committed taxanes biosynthesis in Nicotiana benthamiana

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Jianhua Li ◽  
Ishmael Mutanda ◽  
Kaibo Wang ◽  
Lei Yang ◽  
Jiawei Wang ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Nicotiana Benthamiana ◽  
Functional Expression ◽  
Cytochrome P450 Reductase ◽  
Significant Progress ◽  
Taxadiene Synthase ◽  
Metabolic Engineering Strategy ◽  
Engineering Strategy ◽  
Taxol Production ◽  
Biomass Plant

Abstract Production of the anticancer drug Taxol and its precursors in heterologous hosts is more sustainable than extraction from tissues of yew trees or chemical synthesis. Although attempts to engineer the Taxol pathway in microbes have made significant progress, challenges such as functional expression of plant P450 enzymes remain to be addressed. Here, we introduce taxadiene synthase, taxadiene-5α-hydroxylase, and cytochrome P450 reductase in a high biomass plant Nicotiana benthamiana. Using a chloroplastic compartmentalized metabolic engineering strategy, combined with enhancement of isoprenoid precursors, we show that the engineered plants can produce taxadiene and taxadiene-5α-ol, the committed taxol intermediates, at 56.6 μg g−1 FW and 1.3 μg g−1 FW, respectively. In addition to the tools and strategies reported here, this study highlights the potential of Nicotiana spp. as an alternative platform for Taxol production.

scholarly journals Squalene-Tetrahymanol Cyclase Expression Enables Sterol-Independent Growth of Saccharomyces cerevisiae

2020 ◽  
Vol 86 (17) ◽  
Author(s):  
Sanne J. Wiersma ◽  
Christiaan Mooiman ◽  
Martin Giera ◽  
Jack T. Pronk
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Metabolic Engineering ◽  
Anaerobic Growth ◽  
Sterol Synthesis ◽  
Strictly Anaerobic ◽  
Anaerobic Fungi ◽  
Content Type ◽  
Yeast Strains ◽  
Metabolic Engineering Strategy ◽  
Engineering Strategy

ABSTRACT Biosynthesis of sterols, which are considered essential components of virtually all eukaryotic membranes, requires molecular oxygen. Anaerobic growth of the yeast Saccharomyces cerevisiae therefore strictly depends on sterol supplementation of synthetic growth media. Neocallimastigomycota are a group of strictly anaerobic fungi which, instead of containing sterols, contain the pentacyclic triterpenoid “sterol surrogate” tetrahymanol, which is formed by cyclization of squalene. Here, we demonstrate that expression of the squalene-tetrahymanol cyclase gene TtTHC1 from the ciliate Tetrahymena thermophila enables synthesis of tetrahymanol by S. cerevisiae. Moreover, expression of TtTHC1 enabled exponential growth of anaerobic S. cerevisiae cultures in sterol-free synthetic media. After deletion of the ERG1 gene from a TtTHC1-expressing S. cerevisiae strain, native sterol synthesis was abolished and sustained sterol-free growth was demonstrated under anaerobic as well as aerobic conditions. Anaerobic cultures of TtTHC1-expressing S. cerevisiae on sterol-free medium showed lower specific growth rates and biomass yields than ergosterol-supplemented cultures, while their ethanol yield was higher. This study demonstrated that acquisition of a functional squalene-tetrahymanol cyclase gene offers an immediate growth advantage to S. cerevisiae under anaerobic, sterol-limited conditions and provides the basis for a metabolic engineering strategy to eliminate the oxygen requirements associated with sterol synthesis in yeasts. IMPORTANCE The laboratory experiments described in this report simulate a proposed horizontal gene transfer event during the evolution of strictly anaerobic fungi. The demonstration that expression of a single heterologous gene sufficed to eliminate anaerobic sterol requirements in the model eukaryote Saccharomyces cerevisiae therefore contributes to our understanding of how sterol-independent eukaryotes evolved in anoxic environments. This report provides a proof of principle for a metabolic engineering strategy to eliminate sterol requirements in yeast strains that are applied in large-scale anaerobic industrial processes. The sterol-independent yeast strains described in this report provide a valuable platform for further studies on the physiological roles and impacts of sterols and sterol surrogates in eukaryotic cells.

Metabolic Engineering Strategy Enables a Hundred-Fold Increase in Viniferin Levels in Vitis vinifera cv. Gamay Red Cell Culture

2021 ◽  
Vol 69 (10) ◽  
pp. 3124-3133
Author(s):  
Ru Wang ◽  
Sangram Keshari Lenka ◽  
Varun Kumar ◽  
Kelem Gashu ◽  
Noga Sikron-Persi ◽  
...  
Keyword(s):  
Cell Culture ◽  
Metabolic Engineering ◽  
Vitis Vinifera ◽  
Fold Increase ◽  
Red Cell ◽  
Metabolic Engineering Strategy ◽  
Engineering Strategy
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