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Aleksandra M Mirończuk, Anna Biegalska, Karolina Zugaj, Dorota A Rzechonek, Adam Dobrowolski
Erythritol is a natural sweetener produced by microorganisms as an osmoprotectant. It belongs to the group of polyols and it can be utilized by the oleaginous yeast Yarrowia lipolytica . Despite the recent identification of the transcription factor of erythritol utilization ( EUF1 ), the metabolic pathway of erythritol catabolism remains unknown. In this study we identified a new gene, YALI0F01628g , involved in erythritol assimilation. In silico analysis showed that YALI0F01628g is a putative isomerase and it is localized in the same region as EUF1 ...
2018: Frontiers in Microbiology
Michael Spagnuolo, Murtaza Shabbir Hussain, Lauren Gambill, Mark Blenner
Recent advances in genetic engineering capabilities have enabled the development of oleochemical producing strains of Yarrowia lipolytica . Much of the metabolic engineering effort has focused on pathway engineering of the product using glucose as the feedstock; however, alternative substrates, including various other hexose and pentose sugars, glycerol, lipids, acetate, and less-refined carbon feedstocks, have not received the same attention. In this review, we discuss recent work leading to better utilization of alternative substrates...
2018: Frontiers in Microbiology
Kelly A Markham, Hal S Alper
The oleaginous yeast Yarrowia lipolytica is quickly emerging as the most popular non-conventional (i.e., non-model organism) yeast in the bioproduction field. With a high propensity for flux through tricarboxylic acid (TCA) cycle intermediates and biological precursors such as acetyl-CoA and malonyl-CoA, this host is especially well suited to meet our industrial chemical production needs. Recent progress in synthetic biology tool development has greatly enhanced our ability to rewire this organism, with advances in genetic component design, CRISPR technologies, and modular cloning strategies...
June 4, 2018: Trends in Biotechnology
Erik K R Hanko, Charles M Denby, Violeta Sànchez I Nogué, Weiyin Lin, Kelsey J Ramirez, Christine A Singer, Gregg T Beckham, Jay D Keasling
Medium- and long-chain methyl ketones are fatty acid-derived compounds that can be used as biofuel blending agents, flavors and fragrances. However, their large-scale production from sustainable feedstocks is currently limited due to the lack of robust microbial biocatalysts. The oleaginous yeast Yarrowia lipolytica is a promising biorefinery platform strain for the production of methyl ketones from renewable lignocellulosic biomass due to its natively high flux towards fatty acid biosynthesis. In this study, we report the metabolic engineering of Y...
May 28, 2018: Metabolic Engineering
Difeng Gao, Spencer Smith, Michael Spagnuolo, Gabriel Rodriguez, Mark Blenner
CRISPR-Cas9 technology has been successfully applied in Yarrowia lipolytica for targeted genomic editing including gene disruption and integration; however, disruptions by existing methods typically result from small frameshift mutations caused by indels within the coding region, which usually resulted in unnatural protein. In this study, a dual cleavage strategy directed by paired sgRNAs was developed for gene knockout. This method allows fast and robust gene excision, demonstrated on six genes of interest...
May 29, 2018: Biotechnology Journal
Farshad Darvishi, Mehdi Ariana, Eko Roy Marella, Irina Borodina
Oleaginous yeast Yarrowia lipolytica is an important industrial host for the production of enzymes, oils, fragrances, surfactants, cosmetics, and pharmaceuticals. More recently, improved synthetic biology tools have allowed more extensive engineering of this yeast species, which lead to the production of non-native metabolites. In this review, we summarize the recent advances of genome editing tools for Y. lipolytica, including the application of CRISPR/Cas9 system and discuss case studies, where Y. lipolytica was engineered to produce various non-native chemicals: short-chain fatty alcohols and alkanes as biofuels, polyunsaturated fatty acids for nutritional and pharmaceutical applications, polyhydroxyalkanoates and dicarboxylic acids as precursors for biodegradable plastics, carotenoid-type pigments for food and feed, and campesterol as a precursor for steroid drugs...
May 28, 2018: Applied Microbiology and Biotechnology
Zahra Fathi, Esmail Doustkhah, Sadegh Rostamnia, Farshad Darvishi, Ameneh Ghodsi, Yusuke Ide
Fe3 O4 @C core-shell nanoparticles were modified by (3-aminopropyl)triethoxysilane (APTES) to generate amine functionality in the surface. Then, the amine functional groups were converted to dithiocarbamate via post-modification with carbon disulfide. This nanostructure with new functional property was used to immobilize lipase (obtained from Yarrowia lipolytica U6). Biocatalytic activity of the Fe3 O4 @C-NHCS-LIP was studied in this project. The interaction of lipase and support though dithiocarbamate binder was examined in the hydrolysis of p-nitrophenyl laurate...
May 22, 2018: International Journal of Biological Macromolecules
Lei Qin, Lu Liu, Zhongming Wang, Weining Chen, Dong Wei
To determine the feasibility of microalgae-yeast mixed culture using the liquid digestate of dairy wastewater (LDDW) for biofuels and single cell protein (SCP) production, the cell growth, nutrient removal and outputs evaluation of the mono and mixed culture of Chlorella vulgaris and Yarrowia lipolytica in LDDW were investigated by adding glycerol as carbon source. The results showed that the mixed culture could enhance the biological utilization efficiency of nitrogen and phosphorus, and obtain higher yield of biomass (1...
May 18, 2018: Bioresource Technology
Kurt Patterson, James Yu, Jenny Landberg, Ivan Chang, Farbod Shavarebi, Virginia Bilanchone, Suzanne Sandmeyer
Oleaginous yeasts are valuable systems for biosustainable production of hydrocarbon-based chemicals. Yarrowia lipolytica is one of the best characterized of these yeast with respect to genome annotation and flux analysis of metabolic processes. Nonetheless, progress is hampered by a dearth of genome-wide tools enabling functional genomics. In order to remedy this deficiency, we developed a library of Y. lipolytica insertion mutants via transposon mutagenesis. The Hermes DNA transposon was expressed to achieve saturation mutagenesis of the genome...
May 21, 2018: Metabolic Engineering
Zhong-Peng Guo, Julien Robin, Sophie Duquesne, Michael Joseph O'Donohue, Alain Marty, Florence Bordes
Background: Both industrial biotechnology and the use of cellulosic biomass as feedstock for the manufacture of various commercial goods are prominent features of the bioeconomy. In previous work, with the aim of developing a consolidated bioprocess for cellulose bioconversion, we conferred cellulolytic activity of Yarrowia lipolytica , one of the most widely studied "nonconventional" oleaginous yeast species. However, further engineering this strain often leads to the loss of previously introduced heterologous genes due to the presence of multiple LoxP sites when using Cre -recombinase to remove previously employed selection markers...
2018: Biotechnology for Biofuels
Milda Šulcienė, Boris Kolvenbach, Erik Ammann, Inga Matijošytė
The production of biopolyols (sustainable, renewable and biodegradable precursors), is exclusively focused on the conventional chemical synthesis pathways, and currently, no polyols are produced by enzymatic processes at an industrial scale, because they are not competitive when compared to petrochemically based polyols due to the overall cost for their production. This can be mainly attributed to the high costs of commercial enzymes used in this process and their use is limited by their lack of stability during the bioprocess...
May 16, 2018: International Journal of Biological Macromolecules
James Yu, Jenny Landberg, Farbod Shavarebi, Virginia Bilanchone, Adam Okerlund, Umayangani Wanninayake, Le Zhao, George Kraus, Suzanne Sandmeyer
Yarrowia lipolytica is an oleaginous yeast that is recognized for its ability to accumulate high levels of lipids, which can serve as precursors to biobased fuels and chemicals. Polyketides such as triacetic acid lactone (TAL) can also serve as a precursor for diverse commodity chemicals. This study used Y. lipolytica as a host organism for the production of TAL via expression of the 2-pyrone synthase gene from Gerbera hybrida. Induction of lipid biosynthesis by nitrogen-limited growth conditions increased TAL titers...
May 19, 2018: Biotechnology and Bioengineering
Hui Wang, Zhuang Xue, Zhaoqun Liu, Weilin Wang, Feifei Wang, Ying Wang, Lingling Wang, Linsheng Song
C-type lectins (CTLs) are Ca2+ dependent carbohydrate-binding proteins that share structural homology in their carbohydrate-recognition domains (CRDs). In the present study, a novel CTL was identified from sea cucumber Apostichopus japonicus (named as AjCTL-2). The deduced amino acid sequence of AjCTL-2 was homologous to CTLs from other animals with the identities ranging from 33% to 40%. It contained a canonical signal peptide at the N-terminus, a low density lipoprotein receptor class A (LDLa), a C1r/C1s/Uegf/bone morphogenetic protein 1 (CUB), and a CRD with two motif Glu-Pro-Asn (EPN) and Trp-Asn-Asp (WND) in Ca2+ binding site 2...
May 14, 2018: Fish & Shellfish Immunology
E Carsanba, S Papanikolaou, H Erten
Recently, there has been a great upsurge of interest in studies related to several aspects of microbial lipid production, which is one of the top topics in relevant research fields due to the high demand of these fatty materials in food, medical, oleochemical and biofuel industries. Lipid accumulation by the so-called "oleaginous microorganisms" can generate more than 20% w/w of oil in dry biomass and is governed by a plethora of parameters, such as medium pH, incubation temperature, nutrient limitation and C/N (carbon/nitrogen) ratio, which drastically affect the lipid production bioprocess...
May 15, 2018: Critical Reviews in Biotechnology
Vasiliki Tsakraklides, Annapurna Kamineni, Andrew L Consiglio, Kyle MacEwen, Jonathan Friedlander, Hannah G Blitzblau, Maureen A Hamilton, Donald V Crabtree, Austin Su, Jonathan Afshar, John E Sullivan, W Greg LaTouf, Colin R South, Emily H Greenhagen, A Joe Shaw, Elena E Brevnova
Background: Oleate-enriched triacylglycerides are well-suited for lubricant applications that require high oxidative stability. Fatty acid carbon chain length and degree of desaturation are key determinants of triacylglyceride properties and the ability to manipulate fatty acid composition in living organisms is critical to developing a source of bio-based oil tailored to meet specific application requirements. Results: We sought to engineer the oleaginous yeast Yarrowia lipolytica for production of high-oleate triacylglyceride oil...
2018: Biotechnology for Biofuels
M N Nsoe, G P Kofa, K S Ndi, B Mohammadou, M Heran, G J Kayem
The use of microorganisms for the biodegradation of pollutants is increasingly being studied. But at high concentrations, these pollutants become rather inhibitors for the metabolism of microorganisms. In this study, the biodegradation of ammonium formate at various concentrations (1.59-7.94 mM) by Yarrowia Lipolytica and Pichia guilliermondii isolated from the rubber effluent is studied by following the variation of ammonium ions and formate. A fitting of eight models of substrate inhibition was performed and the parameters were determined by nonlinear regression using MATLAB 8...
2018: Water, Air, and Soil Pollution
Cory Schwartz, Ian Wheeldon
The discovery and adaptation of RNA-guided nucleases has resulted in the rapid development of efficient, scalable, and easily accessible synthetic biology tools for targeted genome editing and transcriptional control. In these systems, for example CRISPR-Cas9 from Streptococcus pyogenes, a protein with nuclease activity is targeted to a specific nucleotide sequence by a short RNA molecule, whereupon binding it cleaves the targeted nucleotide strand. To extend this genome-editing ability to the industrially important oleaginous yeast Yarrowia lipolytica, we developed a set of easily usable and effective CRISPR-Cas9 episomal vectors...
2018: Methods in Molecular Biology
Cory Schwartz, Nicholas Curtis, Ann-Kathrin Löbs, Ian Wheeldon
The yeast Yarrowia lipolytica has been widely studied for its ability to synthesize and accumulate intracellular lipids to high levels. Recent studies have identified native genes that enable growth on biomass-derived sugars, but these genes are not sufficiently expressed to facilitate robust metabolism. In this work, we developed a CRISPR-dCas9 activation (CRISPRa) system in Y. lipolytica and used it to activate native β-glucosidase expression to support growth on cellobiose. A series of different transcriptional activators were compared for their effectiveness in Y...
May 5, 2018: Biotechnology Journal
Man Wang, Guan-Nan Liu, Hong Liu, Lu Zhang, Bing-Zhi Li, Xia Li, Duo Liu, Ying-Jin Yuan
Background: Evolution of complex phenotypes in cells requires simultaneously tuning expression of large amounts of genes, which can be achieved by reprograming global transcription. Lipophilicity is an important complex trait in oleaginous yeast Yarrowia lipolytica . It is necessary to explore the changes of which genes' expression levels will tune cellular lipophilic properties via the strategy of global transcription engineering. Results: We achieved a strategy of global transcription engineering in Y...
2018: Biotechnology for Biofuels
Antonio Milton Vieira Gomes, Talita Souza Carmo, Lucas Silva Carvalho, Frederico Mendonça Bahia, Nádia Skorupa Parachin
Recombinant protein production emerged in the early 1980s with the development of genetic engineering tools, which represented a compelling alternative to protein extraction from natural sources. Over the years, a high level of heterologous protein was made possible in a variety of hosts ranging from the bacteria Escherichia coli to mammalian cells. Recombinant protein importance is represented by its market size, which reached $1654 million in 2016 and is expected to reach $2850.5 million by 2022. Among the available hosts, yeasts have been used for producing a great variety of proteins applied to chemicals, fuels, food, and pharmaceuticals, being one of the most used hosts for recombinant production nowadays...
April 29, 2018: Microorganisms
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