Trends in Oil Production from Oleaginous Yeast Using Biomass: Biotechnological Potential and Constraints

Chaturvedi, Shivani; Bhattacharya, Amrik; Khare, Sunil Kumar

doi:10.1134/s000368381804004x

Cited by 25 publications

(12 citation statements)

References 98 publications

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“…It was observed that the presence of the high percentage of fatty acid (C16-C18 carbon atom in the chain) will seem to be ideal as feedstock for biodiesel [12,17,42]. Producing renewable and environmentally safe biofuel helps in the development of green sustainable energy alternatives [11]. Overexpression and inhibition strategies/metabolic engineering could be identified as one of the efficient methods for increase in biofuel production, which is more convenient and economical compared with molecular gene deletion and addition methodologies.…”

Section: Discussionmentioning

confidence: 99%

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Overexpression and repression of key rate‐limiting enzymes (acetyl CoA carboxylase and HMG reductase) to enhance fatty acid production from Rhodotorula mucilaginosa

et al. 2020

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Implementing two‐way strategies to enhance the lipid production in Rhodotorula mucilaginosa with the help of metabolic engineering was focused on the overexpression of acetyl coenzyme A carboxylase (ACC1 carboxylase) gene and repression of 3‐hydroxy 3‐methylglutaryl reductase (HMG‐CoA reductase). Using an inducer (sodium citrate) and inhibitor (rosuvastatin), the amounts of biomass, lipid, and carotenoid were estimated. In the presence of inhibitor (200 mM), 62% higher lipid concentration was observed, while 44% enhancement was recorded when inducer (3 mM) was used. A combination of both inhibitor and inducer resulted in a 57% increase in lipid concentration by the oleaginous yeast. These results were again confirmed by real‐time polymerase chain reaction by targeting the expression of the genes coding for ACC1 carboxylase and 13‐fold increase was recorded in the presence of inducer as compared with control. This combined strategy (inducer and inhibitor use) has been reported for the first time as far as the best of our knowledge. The metabolic engineering strategies reported here will be a powerful approach for the enhanced commercial production of lipids.

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Section: Discussionmentioning

confidence: 99%

“…| © 2020 Wiley-VCH GmbH agro-waste as a substrate [10,11]. The yeast strains grew robustly on the starchy wastes [12], agro and industrial processing wastes (peel of a banana, potato, yam, cassava and residues of rice, corn, wheat, barley) without any pretreatment, indicating the usefulness of these substrates for the lipid production [2,13,14].…”

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confidence: 98%

Overexpression and repression of key rate‐limiting enzymes (acetyl CoA carboxylase and HMG reductase) to enhance fatty acid production from Rhodotorula mucilaginosa

et al. 2020

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“…27 One of the advantages of oleaginous yeast strains is that they can accumulate intracellular lipids and utilize lignocellulose materials for growth and cell development as well, simultaneously. 28 Recent reviews by Athenaki et al, 29 Chaturvedi et al, 30 Xue et al 31 discussed the physiology of oleaginous yeasts, and their lipids production capability, as well potential biotechnological applications and constraints. It has been reported that Rhodosporidium toruloides and Lipomyces starkeyi can use xylose as a carbon source for cellular growth and can accumulate lipids as well.…”

Section: Oleaginous Yeastmentioning

confidence: 99%

Role of flow cytometry for the improvement of bioprocessing of oleaginous microorganisms

Sonowal

Chikkaputtaiah

Velmurugan

2019

J of Chemical Tech & Biotech

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Oleaginous microorganisms, such as yeast, fungi, microalgae and bacteria, represent a key segment of second generation feed‐stock materials and are considered to synthesize a wide range of industrially important chemical compounds. Oleaginous microorganisms possess a broad varieties of chemical compounds such as carotenoids, pigments, carbohydrates, chlorophyll, and storage‐material lipids. Oleaginous microorganisms have been recognized as promising sources for the synthesis of unsaturated, especially polyunsaturated fatty acids (PUFA). So far, a variety of high‐throughput screening methodologies (HTMs) have been employed for the development of bioprocessing of oleaginous microorganisms for sustainable production of industrially valuable compounds. Of HTMs, flow cytometry (FC) and sorters (FACS) have received substantial interest as better HTMs because of their ability to screen large numbers of cells within seconds, and interrogate and isolate living cells at single‐cell level. Forward and side scattering signals of FC are used to determine the physiological state of the cell while different channels available in the FC facilitate the detection of signals produced from fluorophores. Simultaneous measurement of physiological characteristics along with specific compound accumulation at single‐cell level enables the possibility of separating a particular phenotype with specific properties from a population. Different microbial strain development strategies in combination with FACS produced improved phenotypes with desired properties. This review first summarizes the FACS methodologies suitable for oleaginous microorganisms and the significant progress that has been achieved in oleaginous microorganisms using FACS, and highlights the important, advanced and future prospects of FACS methodologies that are suitable for the development of bioprocessing in oleaginous microorganisms. © 2018 Society of Chemical Industry

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“…The oleaginous yeast can be readily cultivated in high-density cell cultures (1) and naturally accumulates large quantities of both carotenoids and lipids (2) which serve as precursors for many valuable compounds. Complementing this is the ability of R. toruloides to efficiently consume a wide variety of carbon sources, including those found in lignocellulose hydrolysates (3,4). Furthermore, R. toruloides can grow robustly under difficult environmental conditions, including high osmotic stress (5) and the presence of ionic liquids used in pretreatment processes (6)(7)(8).…”

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Multiplexed CRISPR-Cas9-Based Genome Editing of Rhodosporidium toruloides

Otoupal

Ito

Arkin

et al. 2019

mSphere

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Microbial production of biofuels and bioproducts offers a sustainable and economic alternative to petroleum-based fuels and chemicals. The basidiomycete yeast Rhodosporidium toruloides is a promising platform organism for generating bioproducts due to its ability to consume a broad spectrum of carbon sources (including those derived from lignocellulosic biomass) and to naturally accumulate high levels of lipids and carotenoids, two biosynthetic pathways that can be leveraged to produce a wide range of bioproducts. While R. toruloides has great potential, it has a more limited set of tools for genetic engineering relative to more advanced yeast platform organisms such as Yarrowia lipolytica and Saccharomyces cerevisiae. Significant advancements in the past few years have bolstered R. toruloides’ engineering capacity. Here we expand this capacity by demonstrating the first use of CRISPR-Cas9-based gene disruption in R. toruloides. Transforming a Cas9 expression cassette harboring nourseothricin resistance and selecting transformants on this antibiotic resulted in strains of R. toruloides exhibiting successful targeted disruption of the native URA3 gene. While editing efficiencies were initially low (0.002%), optimization of the cassette increased efficiencies 364-fold (to 0.6%). Applying these optimized design conditions enabled disruption of another native gene involved in carotenoid biosynthesis, CAR2, with much greater success; editing efficiencies of CAR2 deletion reached roughly 50%. Finally, we demonstrated efficient multiplexed genome editing by disrupting both CAR2 and URA3 in a single transformation. Together, our results provide a framework for applying CRISPR-Cas9 to R. toruloides that will facilitate rapid and high-throughput genome engineering in this industrially relevant organism. IMPORTANCE Microbial biofuel and bioproduct platforms provide access to clean and renewable carbon sources that are more sustainable and environmentally friendly than petroleum-based carbon sources. Furthermore, they can serve as useful conduits for the synthesis of advanced molecules that are difficult to produce through strictly chemical means. R. toruloides has emerged as a promising potential host for converting renewable lignocellulosic material into valuable fuels and chemicals. However, engineering efforts to improve the yeast’s production capabilities have been impeded by a lack of advanced tools for genome engineering. While this is rapidly changing, one key tool remains unexplored in R. toruloides: CRISPR-Cas9. The results outlined here demonstrate for the first time how effective multiplexed CRISPR-Cas9 gene disruption provides a framework for other researchers to utilize this revolutionary genome-editing tool effectively in R. toruloides.

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Trends in Oil Production from Oleaginous Yeast Using Biomass: Biotechnological Potential and Constraints

Cited by 25 publications

References 98 publications

Overexpression and repression of key rate‐limiting enzymes (acetyl CoA carboxylase and HMG reductase) to enhance fatty acid production from Rhodotorula mucilaginosa

Overexpression and repression of key rate‐limiting enzymes (acetyl CoA carboxylase and HMG reductase) to enhance fatty acid production from Rhodotorula mucilaginosa

Role of flow cytometry for the improvement of bioprocessing of oleaginous microorganisms

Multiplexed CRISPR-Cas9-Based Genome Editing of Rhodosporidium toruloides

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