Yeast and Fungal Metabolites

A special issue of Microorganisms (ISSN 2076-2607). This special issue belongs to the section "Microbial Biotechnology".

Deadline for manuscript submissions: closed (31 January 2020) | Viewed by 46952

Special Issue Editor

Department of Food Science and Human Nutrition, Agricultural University of Athens, Iera Odos 75, 11855 Athens, Greece
Interests: fermentation biotechnology; kinetics and biochemistry of accumulation and degradation of storage lipids by molds and yeasts; biotechnological valorization of agro-industrial wastes and residues; production and study of microbial metabolic products (microbial lipids, citric acid, 1,3-propanediol, enzymes, 2,3-butanediol)
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Special Issue Information

Dear Colleagues,

Molds (fungi that grow in the form of multi-cellular filaments) and yeasts (single-celled fungi) constitute the kingdom of fungi, which includes over 1.0 million different species (only a small portion of these species, viz. c. 10%, have been identified and characterized) and produces a very important plethora of metabolic compounds, both intra-cellular and extra-cellular. In the current Special Issue, research dealing with the production of several types of metabolites that are synthesized by yeasts and fungi and are of interest to the food, chemical, pharmaceutical, and biofuel industries will be presented. Papers dealing with applied biochemistry, fermentation technology, biochemical engineering, and molecular biology related to the synthesis and production of metabolites produced by yeasts and fungi will be considered for publication in this Special Issue. The production aspects of metabolic compounds synthesized by yeasts and fungi that are of particular interest in this Special Issue include, but are not limited to, organic acids (i.e., citric acid, oxalic acid, etc.), polyols (i.e., mannitol, erythritol, arabitol, etc.), ethanol, enzymes, microbial lipids amenable to conversion into second generation biofuels or containing functional poly-unsaturated fatty acids, microbial polysaccharides, etc.

All submitted papers will be subjected to the standard independent peer-review process. Authors should specify “Yeast_Fungal_Metabolites_2019” in the submission cover letter. Prospective authors for mini-reviews or commentaries must contact the Special Issue’s editor and/or the editorial office in advance.

Assoc. Prof. Dr. Seraphim Papanikolaou
Guest Editor

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Published Papers (11 papers)

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Research

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14 pages, 1207 KiB  
Article
Lipids by Yarrowia lipolytica Strains Cultivated on Glucose in Batch Cultures
by Erdem Carsanba, Seraphim Papanikolaou, Patrick Fickers and Huseyin Erten
Microorganisms 2020, 8(7), 1054; https://0-doi-org.brum.beds.ac.uk/10.3390/microorganisms8071054 - 15 Jul 2020
Cited by 36 | Viewed by 4230
Abstract
Oleaginous microorganisms, such as Yarrowia lipolytica, accumulate lipids that can have interesting applications in food biotechnology or the synthesis of biodiesel. Y. lipolytica yeast can have many advantages such as wide substrate range usage and robustness to extreme conditions, while under several [...] Read more.
Oleaginous microorganisms, such as Yarrowia lipolytica, accumulate lipids that can have interesting applications in food biotechnology or the synthesis of biodiesel. Y. lipolytica yeast can have many advantages such as wide substrate range usage and robustness to extreme conditions, while under several culture conditions it can produce high lipid productivity. Based on this assumption, in this study, 12 different Yarrowia lipolytica strains were used to investigate microbial lipid production using a glucose-based medium under nitrogen-limited conditions in shake-flask cultivations. Twelve wild-type or mutant strains of Yarrowia lipolytica which were newly isolated or belonged to official culture collections were tested, and moderate lipid quantities (up to 1.30 g/L) were produced; in many instances, nitrogen limitation led to citric acid production in the medium. Lipids were mainly composed of C16 and C18 fatty acids. Most of the fatty acids of the microbial lipid were unsaturated and corresponded mainly to oleic, palmitic and linoleic acids. Linolenic acid (C18:3) was produced in significant quantities (between 10% and 20%, wt/wt of dry cell weight (DCW)) by strains H917 and Po1dL. Full article
(This article belongs to the Special Issue Yeast and Fungal Metabolites)
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14 pages, 1712 KiB  
Article
Engineering Yarrowia lipolytica for the Synthesis of Glutathione from Organic By-Products
by Diem T. H. Do and Patrick Fickers
Microorganisms 2020, 8(4), 611; https://0-doi-org.brum.beds.ac.uk/10.3390/microorganisms8040611 - 23 Apr 2020
Cited by 4 | Viewed by 3000
Abstract
Tripeptide glutathione, which plays important roles in many cellular mechanisms, is also a biotechnology-oriented molecule with applications in medicine, food and cosmetic. Here, the engineering of the yeast Yarrowia lipolytica for the production of this metabolite at high titer values from various agro-industrial [...] Read more.
Tripeptide glutathione, which plays important roles in many cellular mechanisms, is also a biotechnology-oriented molecule with applications in medicine, food and cosmetic. Here, the engineering of the yeast Yarrowia lipolytica for the production of this metabolite at high titer values from various agro-industrial by-products is reported. The constitutive overexpression of the glutathione biosynthetic genes GSH1 and GSH2 encoding respectively γ-glutamylcysteine synthetase and glutathione synthetase, together with the INU1 gene from Kluyveromyces marxianus encoding inulinase yielded a glutathione titer value and a productivity of 644 nmol/mg protein and 510 µmol/gDCW, respectively. These values were obtained during bioreactor batch cultures in a medium exclusively comprising an extract of Jerusalem artichoke tuber, used as a source of inulin, and ammonium sulfate, used as a nitrogen source. Full article
(This article belongs to the Special Issue Yeast and Fungal Metabolites)
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14 pages, 307 KiB  
Article
Microbiological Production of Isocitric Acid from Biodiesel Waste and Its Effect on Spatial Memory
by Igor G. Morgunov, Svetlana V. Kamzolova, Olga V. Karpukhina, Svetlana B. Bokieva, Julia N. Lunina and Anatoly N. Inozemtsev
Microorganisms 2020, 8(4), 462; https://0-doi-org.brum.beds.ac.uk/10.3390/microorganisms8040462 - 25 Mar 2020
Cited by 6 | Viewed by 2089
Abstract
Within this work, the microbial synthesis of (2R,3S)-isocitric acid (ICA), a metabolite of the nonconventional yeast Yarrowia lipolytica, from biodiesel waste, has been studied. The selected strain Y. lipolytica VKM Y-2373 synthesized ICA with citric acid (CA) as [...] Read more.
Within this work, the microbial synthesis of (2R,3S)-isocitric acid (ICA), a metabolite of the nonconventional yeast Yarrowia lipolytica, from biodiesel waste, has been studied. The selected strain Y. lipolytica VKM Y-2373 synthesized ICA with citric acid (CA) as a byproduct. This process can be regulated by changing cultivation conditions. The maximal production of ICA with the minimal formation of the byproduct was provided by the use of a concentration of (NH4)2SO4 (6 g/L); the addition of biodiesel waste to cultivation medium in 20–60 g/L portions; maintaining the pH of the cultivation medium at 6, and degree of aeration between 25% and 60% of saturation. Itaconic acid at a concentration of 15 mM favorably influenced the production of ICA by the selected strain. The optimization of cultivation conditions allowed us to increase the concentration of ICA in the culture liquid from 58.32 to 90.2 g/L, the product yield (Y) by 40%, and the ICA/CA ratio from 1.1:1 to 3:1. Research on laboratory animals indicated that ICA counteracted the negative effect of ammonium molybdate (10−5 M) and lead diacetate (10−7 M) on the learning and spatial memory of rats, including those exposed to emotional stress. Full article
(This article belongs to the Special Issue Yeast and Fungal Metabolites)
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18 pages, 2051 KiB  
Article
Optimised Production and Extraction of Astaxanthin from the Yeast Xanthophyllomyces dendrorhous
by Zuharlida Tuan Harith, Micael de Andrade Lima, Dimitris Charalampopoulos and Afroditi Chatzifragkou
Microorganisms 2020, 8(3), 430; https://0-doi-org.brum.beds.ac.uk/10.3390/microorganisms8030430 - 19 Mar 2020
Cited by 33 | Viewed by 4703
Abstract
Currently, astaxanthin demand is fulfilled by chemical synthesis using petroleum-based feedstocks. As such, alternative pathways of natural astaxanthin production attracts much research interest. This study aimed at optimising bioreactor operation parameters for astaxanthin production and evaluating strategies for its subsequent extraction. The effect [...] Read more.
Currently, astaxanthin demand is fulfilled by chemical synthesis using petroleum-based feedstocks. As such, alternative pathways of natural astaxanthin production attracts much research interest. This study aimed at optimising bioreactor operation parameters for astaxanthin production and evaluating strategies for its subsequent extraction. The effect of pH and agitation was evident, as a significant reduction in both biomass and astaxanthin production was observed when the culture pH was not controlled and a low agitation speed was applied. At controlled pH conditions and a high agitation speed, a significant increase in biomass (16.4 g/L) and astaxanthin production (3.6 mg/L) was obtained. Enzymatic yeast cell lysis using two commercial enzymes (Accellerase 1500 and Glucanex) was optimised using the central composite design of experiment (DoE). Accellerase 1500 led to mild cell disruption and only 9% (w/w) astaxanthin extraction. However, Glucanex treatment resulted in complete astaxanthin extractability, compared to standard extraction method (DMSO/acetone). When supercritical CO2 was employed as an extraction solvent in Accellerase-pre-treated Xanthophyllomyces dendrorhous cells, astaxanthin extraction increased 2.5-fold. Overall, the study showed that extraction conditions can be tailored towards targeted pigments present in complex mixtures, such as in microbial cells. Full article
(This article belongs to the Special Issue Yeast and Fungal Metabolites)
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14 pages, 739 KiB  
Article
Chemical Composition of an Aphid Antifeedant Extract from an Endophytic Fungus, Trichoderma sp. EFI671
by Nutan Kaushik, Carmen E. Díaz, Hemraj Chhipa, L. Fernando Julio, M. Fe Andrés and Azucena González-Coloma
Microorganisms 2020, 8(3), 420; https://0-doi-org.brum.beds.ac.uk/10.3390/microorganisms8030420 - 17 Mar 2020
Cited by 18 | Viewed by 3224
Abstract
Botanical and fungal biopesticides, including endophytes, are in high demand given the current restrictive legislations on the use of chemical pesticides. As part of an ongoing search for new biopesticides, a series of fungal endophytes have been isolated from selected medicinal plants including [...] Read more.
Botanical and fungal biopesticides, including endophytes, are in high demand given the current restrictive legislations on the use of chemical pesticides. As part of an ongoing search for new biopesticides, a series of fungal endophytes have been isolated from selected medicinal plants including Lauraceae species. In the current study, an extract from the endophytic fungus Trichoderma sp. EFI 671, isolated from the stem parts of the medicinal plant Laurus sp., was screened for bioactivity against plant pathogens (Fusarium graminearum, Rhizoctonia solani, Sclerotinia sclerotiorum and Botrytis cinerea), insect pests (Spodoptera littoralis, Myzus persicae, Rhopalosiphum padi) and plant parasites (Meloidogyne javanica), with positive results against M. persicae. The chemical study of the neutral fraction of the active hexane extract resulted in the isolation of a triglyceride mixture (m1), eburicol (2), β-sitostenone (3), ergosterol (4) and ergosterol peroxide (5). The free fatty acids present in the acid fraction of the extract and in m1 (oleic, linoleic, palmitic and stearic) showed strong dose-dependent antifeedant effects against M. persicae. Liquid (potato dextrose broth, PDB and Sabouraud Broth, SDB) and solid (corn, sorghum, pearl millet and rice) growth media were tested in order to optimize the yield and bioactivity of the fungal extracts. Pearl millet and corn gave the highest extract yields. All the extracts from these solid media had strong effects against M. persicae, with sorghum being the most active. Corn media increased the methyl linoleate content of the extract, pearl millet media increased the oleic acid and sorghum media increased the oleic and linoleic acids compared to rice. The antifeedant effects of these extracts correlated with their content in methyl linoleate and linoleic acid. The phytotoxic effects of these extracts against ryegrass, Lolium perenne, and lettuce, Lactuca sativa, varied with culture media, with sorghum being non- toxic. Full article
(This article belongs to the Special Issue Yeast and Fungal Metabolites)
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12 pages, 2140 KiB  
Article
Identification of Insect-Deterrent Metabolites from Acremonium masseei strain CICY026, a Saprophytic Fungus from a Sinkhole in Yucatán
by Ana L. Ruiz-Jiménez, Esaú Ruiz-Sánchez, Gabriela Heredia, Raúl Tapia-Tussell, Azucena González-Coloma, Karla Peraza-Jiménez, Felicia A. Moo-Koh, Irma L. Medina-Baizabal, Yanet Hernández-Romero, Gonzalo J. Mena-Rejón, Ramiro F. Quijano-Quiñones and Marcela Gamboa-Angulo
Microorganisms 2019, 7(12), 712; https://0-doi-org.brum.beds.ac.uk/10.3390/microorganisms7120712 - 17 Dec 2019
Cited by 4 | Viewed by 2961
Abstract
Micromycetes from unexplored sources represent an opportunity to discover novel natural products to control insect pests. With this aim, a strain of Acremonium masseei CICY026 isolated from a tropical sinkhole was identified, cultured on fermented rice, and its ethyl acetate extract (EAE) was [...] Read more.
Micromycetes from unexplored sources represent an opportunity to discover novel natural products to control insect pests. With this aim, a strain of Acremonium masseei CICY026 isolated from a tropical sinkhole was identified, cultured on fermented rice, and its ethyl acetate extract (EAE) was evaluated against three serious phytophagous insects (Bemisia tabaci, Myzus persicae, and Rhopalosiphum padi). DNA from A. masseei CICY026 was used to confirm its identity. EAE caused settling inhibition (SI) of M. persicae and R. padi (67.5% and 75.3%, respectively). Bioassay-guided fractionation of the active EAE led to the isolation of a novel metabolite, named hexahydroacremonintriol (1), and of acremonin A glucoside (2). The structures of 1 and 2 were determined using IR, one- and two-dimensional NMR, HRMS, and confirmed by theoretical data. The aphid M. persicae was noticeably sensitive to 1 and 2 (SI: 55.6% and 67.2%, respectively), whereas R. padi was only slightly affected by 1 (SI: 59%). This new knowledge about mycobiota from these special sinkhole ecosystems will inform the development of new biorational pesticides. Full article
(This article belongs to the Special Issue Yeast and Fungal Metabolites)
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21 pages, 2756 KiB  
Article
Effect of Myclobutanil Pesticide on the Physiological Behavior of Two Newly Isolated Saccharomyces cerevisiae Strains during Very-High-Gravity Alcoholic Fermentation
by Antonia Terpou, Maria Dimopoulou, Aikaterini Belka, Stamatina Kallithraka, George-John E. Nychas and Seraphim Papanikolaou
Microorganisms 2019, 7(12), 666; https://0-doi-org.brum.beds.ac.uk/10.3390/microorganisms7120666 - 09 Dec 2019
Cited by 15 | Viewed by 2584
Abstract
Yeasts are able to act as biosorbents, as their cell wall includes several components capable of binding organic xenobiotic compounds that can potentially be removed during various fermentation processes. In the present investigation, two novel Saccharomyces cerevisiae strains (LMBF-Y 16 and LMBF-Y-18), previously [...] Read more.
Yeasts are able to act as biosorbents, as their cell wall includes several components capable of binding organic xenobiotic compounds that can potentially be removed during various fermentation processes. In the present investigation, two novel Saccharomyces cerevisiae strains (LMBF-Y 16 and LMBF-Y-18), previously isolated from grapes, were studied regarding their physiological behavior (dry cell weight—DCW production, substrate uptake, and ethanol and glycerol biosynthesis) during fermentations of grape must, in some cases enriched with commercial glucose and fructose (initial total sugar concentration approximately 150 and 250 g/L, respectively). Myclobutanil (a chiral triazole fungicide broadly used as a protective agent of vine) was also added to the culture media at various concentrations in order to assess the ability of the yeasts to simultaneously perform alcoholic fermentations and detoxify the medium (i.e., to remove the fungicide). In the first set of experiments and for both tested strains, trials were carried out in either 250 mL or 2.0 L agitated shake flasks in either synthetic glucose-based experiments or grape musts. Since the results obtained in the trials where the cultures were placed in 2.0 L flasks with grape musts as substrates were superior in terms of both DCW and ethanol production, these experimental conditions were selected for the subsequent studies. Both strains showed high fermentative efficiency, producing high amounts of DCW (9.5–10.5 g/L) in parallel with high ethanol production, which in some cases achieved values very close to the maximum theoretical ethanol production yield (≈0.49 g of ethanol per g of sugar). When using grape must with initial total sugars at approximately 250 g/L (very high gravity fermentation media, close to winemaking conditions), significantly high ethanol quantities (i.e., ranging between 105 and 123 g/L) were produced. Myclobutanil addition slightly negatively affected sugar conversion into ethanol; however, in all cases, ethanol production was very satisfactory. A non-negligible myclobutanil removal during fermentation, which ranged between 5%–27%, as a result of the adsorptive or degradative capacity of the yeast was also reported. The presence of myclobutanil had no effect on DCW production and resulted in no significant differences in the biosynthesis of glycerol. Therefore, these newly isolated yeast strains could be excellent candidates for simultaneous high ethanol production and parallel pesticide removal in a general biorefinery concept demonstrating many environmental benefits. Full article
(This article belongs to the Special Issue Yeast and Fungal Metabolites)
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13 pages, 1230 KiB  
Article
Upgrading Grape Pomace through Pleurotus spp. Cultivation for the Production of Enzymes and Fruiting Bodies
by Aikaterini Papadaki, Vasiliki Kachrimanidou, Seraphim Papanikolaou, Antonios Philippoussis and Panagiota Diamantopoulou
Microorganisms 2019, 7(7), 207; https://0-doi-org.brum.beds.ac.uk/10.3390/microorganisms7070207 - 21 Jul 2019
Cited by 34 | Viewed by 3893
Abstract
Grape pomace, a by-product derived from winery industries, was used as fermentation media for the production of added-value products through the cultivation of two Pleurotus species. Solid-state (SSF), semiliquid (SLF), and submerged (SmF) fermentations were carried out using grape pomace as substrate. The [...] Read more.
Grape pomace, a by-product derived from winery industries, was used as fermentation media for the production of added-value products through the cultivation of two Pleurotus species. Solid-state (SSF), semiliquid (SLF), and submerged (SmF) fermentations were carried out using grape pomace as substrate. The effect of the different fermentations on the consumption of phenolic compounds, the production of mycelial mass and enzymes was evaluated using P. ostreatus and P. pulmonarius. The production of fungal biomass and enzymes was influenced by the fermentation mode. The maximum biomass values of ~0.5 g/g were obtained for both P. pulmonarius and P. ostreatus in SmF. Laccase production was induced in SSF and a maximum activity of 26.247 U/g was determined for P. ostreatus, whereas the highest endoglucanase activity (0.93 U/g) was obtained in the SmF of the same fungi. Analysis of phenolic compounds showed that both strains were able to degrade up to 79% of total phenolic content, regardless the culture conditions. Grape pomace was also evaluated as substrate for mushroom production. P. pulmonarius recorded the highest yield and biological efficiency of 14.4% and 31.4%, respectively. This study showed that mushroom cultivation could upgrade winery by-products towards the production of valuable food products. Full article
(This article belongs to the Special Issue Yeast and Fungal Metabolites)
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Review

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22 pages, 1367 KiB  
Review
Sugar Alcohols and Organic Acids Synthesis in Yarrowia lipolytica: Where Are We?
by Patrick Fickers, Hairong Cheng and Carol Sze Ki Lin
Microorganisms 2020, 8(4), 574; https://0-doi-org.brum.beds.ac.uk/10.3390/microorganisms8040574 - 15 Apr 2020
Cited by 55 | Viewed by 4544
Abstract
Sugar alcohols and organic acids that derive from the metabolism of certain microorganisms have a panoply of applications in agro-food, chemical and pharmaceutical industries. The main challenge in their production is to reach a productivity threshold that allow the process to be profitable. [...] Read more.
Sugar alcohols and organic acids that derive from the metabolism of certain microorganisms have a panoply of applications in agro-food, chemical and pharmaceutical industries. The main challenge in their production is to reach a productivity threshold that allow the process to be profitable. This relies on the construction of efficient cell factories by metabolic engineering and on the development of low-cost production processes by using industrial wastes or cheap and widely available raw materials as feedstock. The non-conventional yeast Yarrowia lipolytica has emerged recently as a potential producer of such metabolites owing its low nutritive requirements, its ability to grow at high cell densities in a bioreactor and ease of genome edition. This review will focus on current knowledge on the synthesis of the most important sugar alcohols and organic acids in Y. lipolytica. Full article
(This article belongs to the Special Issue Yeast and Fungal Metabolites)
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40 pages, 1853 KiB  
Review
An Overview of Potential Oleaginous Microorganisms and Their Role in Biodiesel and Omega-3 Fatty Acid-Based Industries
by Alok Patel, Dimitra Karageorgou, Emma Rova, Petros Katapodis, Ulrika Rova, Paul Christakopoulos and Leonidas Matsakas
Microorganisms 2020, 8(3), 434; https://0-doi-org.brum.beds.ac.uk/10.3390/microorganisms8030434 - 19 Mar 2020
Cited by 144 | Viewed by 12544
Abstract
Microorganisms are known to be natural oil producers in their cellular compartments. Microorganisms that accumulate more than 20% w/w of lipids on a cell dry weight basis are considered as oleaginous microorganisms. These are capable of synthesizing vast majority of fatty acids from [...] Read more.
Microorganisms are known to be natural oil producers in their cellular compartments. Microorganisms that accumulate more than 20% w/w of lipids on a cell dry weight basis are considered as oleaginous microorganisms. These are capable of synthesizing vast majority of fatty acids from short hydrocarbonated chain (C6) to long hydrocarbonated chain (C36), which may be saturated (SFA), monounsaturated (MUFA), or polyunsaturated fatty acids (PUFA), depending on the presence and number of double bonds in hydrocarbonated chains. Depending on the fatty acid profile, the oils obtained from oleaginous microorganisms are utilized as feedstock for either biodiesel production or as nutraceuticals. Mainly microalgae, bacteria, and yeasts are involved in the production of biodiesel, whereas thraustochytrids, fungi, and some of the microalgae are well known to be producers of very long-chain PUFA (omega-3 fatty acids). In this review article, the type of oleaginous microorganisms and their expertise in the field of biodiesel or omega-3 fatty acids, advances in metabolic engineering tools for enhanced lipid accumulation, upstream and downstream processing of lipids, including purification of biodiesel and concentration of omega-3 fatty acids are reviewed. Full article
(This article belongs to the Special Issue Yeast and Fungal Metabolites)
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11 pages, 732 KiB  
Review
Genomic Considerations for the Modification of Saccharomyces cerevisiae for Biofuel and Metabolite Biosynthesis
by James T. Arnone
Microorganisms 2020, 8(3), 321; https://0-doi-org.brum.beds.ac.uk/10.3390/microorganisms8030321 - 26 Feb 2020
Cited by 14 | Viewed by 2390
Abstract
The growing global population and developing world has put a strain on non-renewable natural resources, such as fuels. The shift to renewable sources will, thus, help meet demands, often through the modification of existing biosynthetic pathways or the introduction of novel pathways into [...] Read more.
The growing global population and developing world has put a strain on non-renewable natural resources, such as fuels. The shift to renewable sources will, thus, help meet demands, often through the modification of existing biosynthetic pathways or the introduction of novel pathways into non-native species. There are several useful biosynthetic pathways endogenous to organisms that are not conducive for the scale-up necessary for industrial use. The use of genetic and synthetic biological approaches to engineer these pathways in non-native organisms can help ameliorate these challenges. The budding yeast Saccharomyces cerevisiae offers several advantages for genetic engineering for this purpose due to its widespread use as a model system studied by many researchers. The focus of this review is to present a primer on understanding genomic considerations prior to genetic modification and manipulation of S. cerevisiae. The choice of a site for genetic manipulation can have broad implications on transcription throughout a region and this review will present the current understanding of position effects on transcription. Full article
(This article belongs to the Special Issue Yeast and Fungal Metabolites)
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