Functional characterization and overexpression of Δ12-desaturase in the oleaginous yeast Rhodotorula toruloides for production of linoleic acid-rich lipids

Wu, Chih-Chan; Ohashi, Takao; Kajiura, Hiroyuki; Sato, Yu; Misaki, Ryo; Honda, Kohsuke; Fujiyama, Kazuhito

doi:10.1016/j.jbiosc.2021.02.002

Cited by 17 publications

(13 citation statements)

References 38 publications

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“…Lamers and co-workers [ 3 ] increased the LA content (from 5.6% to 23.3% and 22.7%) in Schwanniomyces occidentalis by homologously and heterologously expressing FAD2. Subsequently, Δ12-fatty acid desaturase was successfully expressed in Rhodotorula toruloides and improved the final titer of LA to 1.3 g/L [ 17 ]. LA is a precursor and limiting factor for PUFA biosynthesis in M. circinelloides , since there is only one Δ12-desaturase gene in its genome, and the LA content is approximately 15 % of the total lipids [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Homologous and Heterologous Expression of Delta(12)-Desaturase in Mucor circinelloides Enhanced the Production of Linolenic Acid

et al. 2022

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Linolenic acid (LA) is gaining more interest within the scientific community. This is because it has a potential medical role in reducing the risk of inflammation, carcinogenesis, atherosclerosis and diabetes and is a valuable nutraceutical for human health. The oleaginous fungus Mucor circinelloides produces a high lipid content (36%), including valuable polyunsaturated fatty acids (PUFAs). However, the critical step in which oleic acid (OA) is converted into LA is not efficient at supplying enough substrates for PUFA synthesis. Hence, we propose a method to increase LA production based on genetic engineering. The overexpression of the Δ12-desaturase gene from M. circinelloides and Mortierella alpina increased the LA content and improved the lipid accumulation (from 14.9% to 21.6% in the Δ12-desaturase gene of the M. circinelloides overexpressing strain (Mc-D12MC) and from 14.9% to 18.7% in the Δ12-desaturase gene of M. alpina overexpressing strain (Mc-D12MA)). Additionally, the up-regulated expression levels of these genes targeted the genes involved in NADPH production, implying that the elevated Δ12-desaturase gene may function as a critical regulator of NADPH and lipid synthesis in M. circinelloides. This study provides the first evidence to support the design of metabolic engineering related to LA and PUFA production in M. circinelloides for potential industrial applications.

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Section: Introductionmentioning

confidence: 99%

Homologous and Heterologous Expression of Delta(12)-Desaturase in Mucor circinelloides Enhanced the Production of Linolenic Acid

et al. 2022

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“…After heterologous overexpress Δ12-desaturase from M. alpina or Fusarium verticillioides in the oleaginous yeast Rhodosporidium toruloides , linoleic acid concentration increased up to 1.3 g/L, which was 5-fold higher than that in the parent strain ( 61 ). Overexpression of endogenous RtFAD2 in R. toruloides also improved lipid and linoleic acid ( 32 ). Conversely, the growth rate was slower at 12°C under the deletion of FAD2 gene (coding Δ12-desaturase) in Y. lipolytica , which was recovered by the addition of 18:2, not 18:1.…”

Section: Vital Desaturase In ω-6 and ω-3 Pathwaymentioning

confidence: 99%

Key Enzymes in Fatty Acid Synthesis Pathway for Bioactive Lipids Biosynthesis

et al. 2022

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Dietary bioactive lipids, one of the three primary nutrients, is not only essential for growth and provides nutrients and energy for life's activities but can also help to guard against disease, such as Alzheimer's and cardiovascular diseases, which further strengthen the immune system and maintain many body functions. Many microorganisms, such as yeast, algae, and marine fungi, have been widely developed for dietary bioactive lipids production. These biosynthetic processes were not limited by the climate and ground, which are also responsible for superiority of shorter periods and high conversion rate. However, the production process was also exposed to the challenges of low stability, concentration, and productivity, which was derived from the limited knowledge about the critical enzyme in the metabolic pathway. Fortunately, the development of enzymatic research methods provides powerful tools to understand the catalytic process, including site-specific mutagenesis, protein dynamic simulation, and metabolic engineering technology. Thus, we review the characteristics of critical desaturase and elongase involved in the fatty acids' synthesis metabolic pathway, which aims to not only provide extensive data for enzyme rational design and modification but also provides a more profound and comprehensive understanding of the dietary bioactive lipids' synthetic process.

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“…The importance of fatty acid desaturases as potential targets for increasing lipid accumulation was widely examined. The expression of these membrane-bound proteins that catalyse the addition of a double bond in fatty acid hydrocarbon chains to produce unsaturated and polyunsaturated fatty acids, was found to enhance lipid production in several yeast species/strains as well as the accumulation of unsaturated lipids, which is beneficial for biodiesel production [ 223 , 225 , 242 , 243 , 244 ].…”

Section: Strategies To Develop Superior Strains For the Production Of...mentioning

confidence: 99%

“…For this reason, the majority of membrane engineering attempts to increase tolerance to multiple stresses target the modulation of its lipid composition, in order to maintain the integrity and fluidity under stress, namely by altering lipid saturation or changing the length of lipid in biomembranes [ 264 ]. The genetic manipulation of oleaginous yeasts, comprising the degree of saturation of lipids [ 200 , 223 , 243 , 244 , 270 , 271 , 272 ] or the length of the lipidic chain [ 273 , 274 ] led to increased lipid titers. Additionally, membrane proteins including integral membrane proteins and transport proteins are also extremely relevant in stress tolerance.…”

Section: Strategies To Develop Superior Strains For the Production Of...mentioning

confidence: 99%

Exploring Yeast Diversity to Produce Lipid-Based Biofuels from Agro-Forestry and Industrial Organic Residues

Mota

Múgica²,

Correia

2022

JoF

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Exploration of yeast diversity for the sustainable production of biofuels, in particular biodiesel, is gaining momentum in recent years. However, sustainable, and economically viable bioprocesses require yeast strains exhibiting: (i) high tolerance to multiple bioprocess-related stresses, including the various chemical inhibitors present in hydrolysates from lignocellulosic biomass and residues; (ii) the ability to efficiently consume all the major carbon sources present; (iii) the capacity to produce lipids with adequate composition in high yields. More than 160 non-conventional (non-Saccharomyces) yeast species are described as oleaginous, but only a smaller group are relatively well characterised, including Lipomyces starkeyi, Yarrowia lipolytica, Rhodotorula toruloides, Rhodotorula glutinis, Cutaneotrichosporon oleaginosus and Cutaneotrichosporon cutaneum. This article provides an overview of lipid production by oleaginous yeasts focusing on yeast diversity, metabolism, and other microbiological issues related to the toxicity and tolerance to multiple challenging stresses limiting bioprocess performance. This is essential knowledge to better understand and guide the rational improvement of yeast performance either by genetic manipulation or by exploring yeast physiology and optimal process conditions. Examples gathered from the literature showing the potential of different oleaginous yeasts/process conditions to produce oils for biodiesel from agro-forestry and industrial organic residues are provided.

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Functional characterization and overexpression of Δ12-desaturase in the oleaginous yeast Rhodotorula toruloides for production of linoleic acid-rich lipids

Cited by 17 publications

References 38 publications

Homologous and Heterologous Expression of Delta(12)-Desaturase in Mucor circinelloides Enhanced the Production of Linolenic Acid

Homologous and Heterologous Expression of Delta(12)-Desaturase in Mucor circinelloides Enhanced the Production of Linolenic Acid

Key Enzymes in Fatty Acid Synthesis Pathway for Bioactive Lipids Biosynthesis

Exploring Yeast Diversity to Produce Lipid-Based Biofuels from Agro-Forestry and Industrial Organic Residues

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