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New Perspectives for Oleaginous Fungi
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New Perspectives for Oleaginous Fungi

A special issue of Journal of Fungi (ISSN 2309-608X). This special issue belongs to the section "Fungal Genomics, Genetics and Molecular Biology".

Deadline for manuscript submissions: closed (30 April 2023) | Viewed by 12726

Special Issue Editors

Prof. Dr. Yuanda Song

E-Mail Website
Guest Editor
Director of Colin Ratledge Center for Microbial Lipids, School of Agriculture Engineering and Food Science, Shandong University of Technology, 266 Xincun Rd., Zibo, China
Interests: molecular mechanisms of lipid biosynthesis; biochemistry; microbial lipids; M. circinelloides; genetic engineering; fatty acids synthesis; biomass and value-added products; chemical engineering; fermentation biotechnology; food chemistry; functional foods; applied microbiology; natural products
Prof. Dr. Milan Čertík

E-Mail Website
Guest Editor
Department of Biochemical Technology, Faculty of Chemical and Food Technology, Institute of Biotechnology, Slovak University of Technology in Bratislava, Bratislava, Slovakia
Interests: industrial biotechnology; applied microbiology; biochemistry; agricultural biotechnology; microbial biotechnology; fatty acids; lipophilic compounds; genetic engineering; lipid metabolism; metabolic engineering; fermentation biotechnology

Special Issue Information

Dear Colleagues,

Nowadays, there is considerable concern over the energy shortage and nutritional problems accompanying the rapid growth of the world population and economic situation. Thus, it is necessary to find suitable and cheap sources for lipid production. Oleaginous fungi and yeasts have been considered potential oil sources for lipid and biodiesel production based on their ability to accumulate large amounts of lipids. However, to develop efficient technologies for lipid and biofuels production, significant research needs to be directed towards biotechnological and genetic improvement of the potential organisms utilized in the process. Despite the diversity of oil-bearing fungi, only a few oleaginous fungal models have been well recognized to study their lipid production. Mortierella alpina, Mucor circinelloides, Umbelopsis isabellina, Cunninghamella echinulata, and Yarrowia lipolytica represent the well-established models, while new effective molecular and genetic tools have successfully contributed to engaging with additional oleaginous fungal model oleaginous.

In this Special Issue, we will cover some recent progress focused on novel molecular techniques and the development of additional fungal models for varied biotechnological applications and their ability to modulate, regulate, and maximize production. We aim to (a) provide an introductory overview of fungal lipids and show the current interest in their production and use the oleaginous fungi as sources of oils and fats, (b) high throughput screening technologies for bioactive lipid and fatty acids producing fungi, (c) highlight novel methods and trends proposed to produce lipids and lipid-derivatives in sufficient quantities from oleaginous fungi, (d) provides an overview on discovered model species, (e) give insights into the diversity of molecular biological manipulations and genetics that are useful for biotechnological application for lipid production.

Prof. Dr. Yuanda Song
Prof. Dr. Milan Čertík
Guest Editors

Manuscript Submission Information

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Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • oleaginous fungi
  • fungal lipids
  • lipid biosynthesis
  • fermentation
  • genetic manipulation
  • lipid biochemistry
  • fatty acids
  • lipid accumulation
  • biofuels
  • biotechnology

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Published Papers (6 papers)

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Research

30 pages, 8480 KiB  
Article
Poly-Unsaturated Fatty Acids (PUFAs) from Cunninghamella elegans Grown on Glycerol Induce Cell Death and Increase Intracellular Reactive Oxygen Species
by Georgios Kalampounias, Chrysavgi Gardeli, Spyridon Alexis, Elena Anagnostopoulou, Theodosia Androutsopoulou, Panagiotis Dritsas, George Aggelis, Seraphim Papanikolaou and Panagiotis Katsoris
J. Fungi 2024, 10(2), 130; https://doi.org/10.3390/jof10020130 - 4 Feb 2024
Cited by 1 | Viewed by 2389
Abstract
Cunninghamella elegans NRRL-1393 is an oleaginous fungus able to synthesize and accumulate unsaturated fatty acids, amongst which the bioactive gamma-linolenic acid (GLA) has potential anti-cancer activities. C. elegans was cultured in shake-flask nitrogen-limited media with either glycerol or glucose (both at ≈60 g/L) [...] Read more.
Cunninghamella elegans NRRL-1393 is an oleaginous fungus able to synthesize and accumulate unsaturated fatty acids, amongst which the bioactive gamma-linolenic acid (GLA) has potential anti-cancer activities. C. elegans was cultured in shake-flask nitrogen-limited media with either glycerol or glucose (both at ≈60 g/L) employed as the sole substrate. The assimilation rate of both substrates was similar, as the total biomass production reached 13.0–13.5 g/L, c. 350 h after inoculation (for both instances, c. 27–29 g/L of substrate were consumed). Lipid production was slightly higher on glycerol-based media, compared to the growth on glucose (≈8.4 g/L vs. ≈7.0 g/L). Lipids from C. elegans grown on glycerol, containing c. 9.5% w/w of GLA, were transformed into fatty acid lithium salts (FALS), and their effects were assessed on both human normal and cancerous cell lines. The FALS exhibited cytotoxic effects within a 48 h interval with an IC50 of about 60 μg/mL. Additionally, a suppression of migration was shown, as a significant elevation of oxidative stress levels, and the induction of cell death. Elementary differences between normal and cancer cells were not shown, indicating a generic mode of action; however, oxidative stress level augmentation may increase susceptibility to anticancer drugs, improving chemotherapy effectiveness. Full article
(This article belongs to the Special Issue New Perspectives for Oleaginous Fungi)
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13 pages, 2992 KiB  
Article
Applications of Diacylglycerol Acyltransferase for Triacylglycerol Production in Mortierella alpina
by Ruilin Yu, Lulu Chang, Jun Cao, Bo Yang, Haiqin Chen and Wei Chen
J. Fungi 2023, 9(2), 219; https://doi.org/10.3390/jof9020219 - 7 Feb 2023
Viewed by 1388
Abstract
Triacylglycerol (TG) with high-value long-chain polyunsaturated fatty acids is beneficial to human health; consequently, there is an urgent need to broaden its sources due to the current growing demand. Mortierella alpina, one of the most representative oleaginous fungi, is the only certificated [...] Read more.
Triacylglycerol (TG) with high-value long-chain polyunsaturated fatty acids is beneficial to human health; consequently, there is an urgent need to broaden its sources due to the current growing demand. Mortierella alpina, one of the most representative oleaginous fungi, is the only certificated source of dietary arachidonic acid-rich oil supplied in infant formula. This study was conducted to improve TG production in M. alpina by homologous overexpression of diacylglycerol acyltransferase (DGAT) and linseed oil (LSO) supplementation. Our results showed that the homologous overexpression of MaDGAT1B and MaDGAT2A strengthened TG biosynthesis and significantly increased the TG content compared to the wild-type by 12.24% and 14.63%, respectively. The supplementation with an LSO concentration of 0.5 g/L elevated the TG content to 83.74% and total lipid yield to 4.26 ± 0.38 g/L in the M. alpina-MaDGAT2A overexpression strain. Our findings provide an effective strategy for enhancing TG production and highlight the role of DGAT in TG biosynthesis in M. alpina. Full article
(This article belongs to the Special Issue New Perspectives for Oleaginous Fungi)
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22 pages, 28446 KiB  
Article
Insights into the Genomic and Phenotypic Landscape of the Oleaginous Yeast Yarrowia lipolytica
by Frédéric Bigey, Emilie Pasteur, Xymena Połomska, Stéphane Thomas, Anne-Marie Crutz-Le Coq, Hugo Devillers and Cécile Neuvéglise
J. Fungi 2023, 9(1), 76; https://doi.org/10.3390/jof9010076 - 4 Jan 2023
Cited by 1 | Viewed by 2711
Abstract
Although Yarrowia lipolytica is a model yeast for the study of lipid metabolism, its diversity is poorly known, as studies generally consider only a few standard laboratory strains. To extend our knowledge of this biotechnological workhorse, we investigated the genomic and phenotypic diversity [...] Read more.
Although Yarrowia lipolytica is a model yeast for the study of lipid metabolism, its diversity is poorly known, as studies generally consider only a few standard laboratory strains. To extend our knowledge of this biotechnological workhorse, we investigated the genomic and phenotypic diversity of 56 natural isolates. Y. lipolytica is classified into five clades with no correlation between clade membership and geographic or ecological origin. A low genetic diversity (π = 0.0017) and a pan-genome (6528 genes) barely different from the core genome (6315 genes) suggest Y. lipolytica is a recently evolving species. Large segmental duplications were detected, totaling 892 genes. With three new LTR-retrotransposons of the Gypsy family (Tyl4, Tyl9, and Tyl10), the transposable element content of genomes appeared diversified but still low (from 0.36% to 3.62%). We quantified 34 traits with substantial phenotypic diversity, but genome-wide association studies failed to evidence any associations. Instead, we investigated known genes and found four mutational events leading to XPR2 protease inactivation. Regarding lipid metabolism, most high-impact mutations were found in family-belonging genes, such as ALK or LIP, and therefore had a low phenotypic impact, suggesting that the huge diversity of lipid synthesis and accumulation is multifactorial or due to complex regulations. Full article
(This article belongs to the Special Issue New Perspectives for Oleaginous Fungi)
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12 pages, 2837 KiB  
Article
Heterologous Expression of CFL1 Confers Flocculating Ability to Cutaneotrichosporon oleaginosus Lipid-Rich Cells
by Silvia Donzella and Concetta Compagno
J. Fungi 2022, 8(12), 1293; https://doi.org/10.3390/jof8121293 - 11 Dec 2022
Viewed by 1600
Abstract
Lipid extraction from microbial and microalgae biomass requires the separation of oil-rich cells from the production media. This downstream procedure represents a major bottleneck in biodiesel production, increasing the cost of the final product. Flocculation is a rapid and cheap system for removing [...] Read more.
Lipid extraction from microbial and microalgae biomass requires the separation of oil-rich cells from the production media. This downstream procedure represents a major bottleneck in biodiesel production, increasing the cost of the final product. Flocculation is a rapid and cheap system for removing solid particles from a suspension. This natural characteristic is displayed by some microorganisms due to the presence of lectin-like proteins (called flocculins/adhesins) in the cell wall. In this work, we showed, for the first time, that the heterologous expression of the adhesin Cfl1p endows the oleaginous species Cutaneotrichosporon oleaginosus with the capacity of cell flocculation. We used Helm’s test to demonstrate that the acquisition of this trait allows for reducing the time required for the separation of lipid-rich cells from liquid culture by centrifugation without altering the productivity. This improves the lipid production process remarkably by providing a more efficient downstream. Full article
(This article belongs to the Special Issue New Perspectives for Oleaginous Fungi)
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10 pages, 1068 KiB  
Article
Investigating the Effect of Alcohol Dehydrogenase Gene Knockout on Lipid Accumulation in Mucor circinelloides WJ11
by Aabid Manzoor Shah, Hassan Mohamed, Abu Bakr Ahmad Fazili, Wu Yang and Yuanda Song
J. Fungi 2022, 8(9), 917; https://doi.org/10.3390/jof8090917 - 29 Aug 2022
Cited by 6 | Viewed by 1814
Abstract
Mucor circinelloides is an oleaginous, dimorphic zygomycete fungus species that produces appreciable levels of ethanol when grown under aerobic conditions in the presence of high glucose, indicating the fungus is a Crabtree-positive microorganism. Engineering efforts to redirect carbon flux from ethanol to lipid [...] Read more.
Mucor circinelloides is an oleaginous, dimorphic zygomycete fungus species that produces appreciable levels of ethanol when grown under aerobic conditions in the presence of high glucose, indicating the fungus is a Crabtree-positive microorganism. Engineering efforts to redirect carbon flux from ethanol to lipid biosynthesis may shed light on the critical role of ethanol biosynthesis during aerobic fermentation in M. circinelloides. Therefore, in this study, the alcohol dehydrogenase gene (ADH1) of M. circinelloides WJ11 was deleted, and its effects on growth, lipid production, and fatty acid content were analyzed. Our results showed that knocking out of adh1∆ reduced the ethanol concentration by 85–90% in fermented broth, indicating that this gene is the major source of ethanol production. Parallel to these findings, the lipid and fatty acid content of the mutant was decreased, while less change in the growth of WJ11 was observed. Furthermore, a fermentation study showed the lipid and fatty acid content was restored in the mutant strain when the fermentation media was supplemented with 0.5% external ethanol, indicating the importance of alcohol dehydrogenase and its product on growth and lipid biosynthesis in M. circinelloides. To our knowledge, this is the first study to show a link between alcohol dehydrogenase and lipid production in M. circinelloides. Full article
(This article belongs to the Special Issue New Perspectives for Oleaginous Fungi)
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13 pages, 2689 KiB  
Article
Comparative Genomics of Mortierellaceae Provides Insights into Lipid Metabolism: Two Novel Types of Fatty Acid Synthase
by Heng Zhao, Yong Nie, Yang Jiang, Shi Wang, Tian-Yu Zhang and Xiao-Yong Liu
J. Fungi 2022, 8(9), 891; https://doi.org/10.3390/jof8090891 - 23 Aug 2022
Cited by 5 | Viewed by 2038
Abstract
Fungal species in the family Mortierellaceae are important for their remarkable capability to synthesize large amounts of polyunsaturated fatty acids, especially arachidonic acid (ARA). Although many genomes have been published, the quality of these data is not satisfactory, resulting in an incomplete understanding [...] Read more.
Fungal species in the family Mortierellaceae are important for their remarkable capability to synthesize large amounts of polyunsaturated fatty acids, especially arachidonic acid (ARA). Although many genomes have been published, the quality of these data is not satisfactory, resulting in an incomplete understanding of the lipid pathway in Mortierellaceae. We provide herein two novel and high-quality genomes with 55.32% of syntenic gene pairs for Mortierella alpina CGMCC 20262 and M. schmuckeri CGMCC 20261, spanning 28 scaffolds of 40.22 Mb and 25 scaffolds of 49.24 Mb, respectively. The relative smaller genome for the former is due to fewer protein-coding gene models (11,761 vs. 13,051). The former yields 45.57% of ARA in total fatty acids, while the latter 6.95%. The accumulation of ARA is speculated to be associated with delta-5 desaturase (Delta5) and elongation of very long chain fatty acids protein 3 (ELOVL3). A further genomic comparison of 19 strains in 10 species in three genera in the Mortierellaceae reveals three types of fatty acid synthase (FAS), two of which are new to science. The most common type I exists in 16 strains of eight species of three genera, and was discovered previously and consists of a single unit with eight active sites. The newly revealed type II exists only in M. antarctica KOD 1030 where the unit is separated into two subunits α and β comprised of three and five active sites, respectively. Another newly revealed type III exists in M. alpina AD071 and Dissophora globulifera REB-010B, similar to type II but different in having one more acyl carrier protein domain in the α subunit. This study provides novel insights into the enzymes related to the lipid metabolism, especially the ARA-related Delta5, ELOVL3, and FAS, laying a foundation for genetic engineering of Mortierellaceae to modulate yield in polyunsaturated fatty acids. Full article
(This article belongs to the Special Issue New Perspectives for Oleaginous Fungi)
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