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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
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
Lubna Richter, Cresten B Mansfeldt, Michael M Kuan, Alexandra Cesare, Stephen T Menefee, Ruth E Richardson, Beth A Ahner
Given the challenges facing the economically favorable production of products from microalgae, understanding factors that might impact productivity rates including growth rates and accumulation of desired products, e.g., triacylglycerols (TAG) for biodiesel feedstock, remains critical. Although operational parameters such as media composition and reactor design can clearly effect growth rates, the role of microbe-microbe interactions is just beginning to be elucidated. In this study an oleaginous marine algae Chlorella spp...
May 11, 2018: Environmental Science & Technology
Madan Kumar Mandal, Neha Chaurasia
Biomass and lipid yield under nutrient depleted and supplemented conditions of N and Mg in two freshwater isolates, Chlorella sp. NC-MKM and Scenedesmus acutus NC-M2 from Meghalaya were investigated for biodiesel production. Both the strains, Chlorella sp. NC-MKM and Scenedesmus acutus NC-M2 are oleaginous in nature having lipid content of 40.2 ± 1.4 and 37.3 ± 2.6% DCW, respectively. The significant increase (92.8%) in lipid content was found in N-depleted condition while an increase (46.65%) in biomass yield was observed under Mg-supplemented condition in Chlorella sp...
May 2018: 3 Biotech
Mohammad Enamul Haque, Bing Han, Bin Wang, Yue Wang, Aizhong Liu
Chromatin immunoprecipitation (ChIP) is usually a reliable technique to find the binding sites of a transcription factor. In the current study, we developed a suitable ChIP method using developing castor bean seeds. A castor bean seed with large and persistent endosperm contains high amounts of storage lipids (ca. 50-60%) and is often considered as a model material to studying seed biology. In oleaginous seeds, due to the rich oils which could seriously affect immunoprecipitation and DNA isolation, it is often difficult to carry out a successful ChIP experiment...
2018: PloS One
Zhongjie Wang, Xiaobin Wen, Yan Xu, Yi Ding, Yahong Geng, Yeguang Li
Carbon dioxide (CO2 ) and pH are two interdependent factors that greatly impact the growth and lipid accumulation of microalgae. However, the effects of these two factors are usually studied separately. The use of exogenous CO2 , such as flue gas derived, to regulate pH in the large-scale cultivation of microalgae provides an ideal means for combining CO2 biofixation and biodiesel production. In this study, the CO2 biofixation and lipid production of oleaginous microalga Graesiella sp. WBG-1 was explored for four pH levels regulated by exogenous 15% CO2 (flue gas concentration) in 10L circular culture ponds and 5m2 open raceway reactors...
April 1, 2018: Science of the Total Environment
Zhu Chen, Wesley D Reznicek, Caixia Wan
In this study, an acidified, aqueous DES comprising choline chloride: glycerol (ChCl:Gly) was used to fractionate switchgrass into three major streams under a relatively mild condition: cellulose-rich pulp, lignin, and xylose-rich liquor. The pulp showed good digestibility with about 89% glucose yield. The solvent can be recycled successfully and reused for at least four more pretreatment cycles while maintaining its pretreatment capability. The solvent recycling also improved the lignin recovery from the pretreatment liquor...
April 18, 2018: Bioresource Technology
P Selvakumar, P Sivashanmugam
The economical production of lipids is considered as an appropriate renewable alternative feedstock for biodiesel production because of the contemporary concerns on fuel crisis, climate change and food security. In this study, lipid accumulation potential of a novel oleaginous yeast isolate Naganishia liquefaciens NITTS2 by utilizing pre-digested municipal waste activated sludge (PWAS) was explored. Optimization of culture conditions was performed using response surface methodology coupled with genetic algorithm and maximum lipid content of 55...
May 5, 2018: Applied Biochemistry and Biotechnology
Tatsuhiro Nomaguchi, Yoshiaki Maeda, Tomoko Yoshino, Toru Asahi, Leila Tirichine, Chris Bowler, Tsuyoshi Tanaka
BACKGROUND: Allopolyploidy is a genomic structure wherein two or more sets of chromosomes derived from divergent parental species coexist within an organism. It is a prevalent genomic configuration in plants, as an important source of genetic variation, and also frequently confers environmental adaptability and increased crop productivity. We previously reported the oleaginous marine diatom Fistulifera solaris JPCC DA0580 to be a promising host for biofuel production and that its genome is allopolyploid, which had never previously been reported in eukaryotic microalgae...
May 4, 2018: BMC Genomics
Batul Diwan, Pratima Gupta
AIMS: Oleaginous yeasts were isolated from five different rotten fruits and their oleaginous behaviour was characterized with varying carbon source and concentration (C/N) to pave the path for their future applicabilities. METHODS & RESULTS: Sixteen yeasts were isolated, those accumulating around 20% of their cell dry weights as lipids were screened out and identified as Candida tropicalis and Pichia kudriavzevii. Amongst glucose, maltose, and sucrose, glucose was found favorable carbon source in terms of biomass yield and lipid content...
May 4, 2018: Journal of Applied Microbiology
Hyun Gi Koh, Nam Kyu Kang, Eun Kyung Kim, Seungjib Jeon, Sung-Eun Shin, Bongsoo Lee, Yong Keun Chang
Even though there has been much interest in genetic engineering of microalgae, its progress has been slow due to the difficulty and limitation of available techniques. Currently, genetic modification in most microalgal strains is confined to single gene transformation. Here, a multigene expression system for the oleaginous model strain Nannochloropsis salina was developed with glycine-serine-glycine spacer linked 2A self-cleaving peptides (2A) for the first time. An efficiency test of the four most widely used 2As revealed that two different types of 2As T2A and E2A have the best performance in N...
April 28, 2018: Journal of Biotechnology
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
Qinhua Gan, Jiaoyun Jiang, Xiao Han, Shifan Wang, Yandu Lu
Plastid engineering offers an important tool to fill the gap between the technical and the enormous potential of microalgal photosynthetic cell factory. However, to date, few reports on plastid engineering in industrial microalgae have been documented. This is largely due to the small cell sizes and complex cell-wall structures which make these species intractable to current plastid transformation methods (i.e., biolistic transformation and polyethylene glycol-mediated transformation). Here, employing the industrial oleaginous microalga Nannochloropsis oceanica as a model, an electroporation-mediated chloroplast transformation approach was established...
2018: Frontiers in Plant Science
Qi Gao, Xuan Cao, Yuying Huang, Jinglin Yang, Jun Chen, Liujing Wei, Qiang Hua
Recent advances in the production of biofuels by microbes have attracted attention due to increasingly-limited fossil fuels. Biodiesels, especially fatty acid ethyl esters (FAEEs), are considered a potentially fully-sustainable fuel in the near future due to similarities with petrodiesels and compatibility with existing infrastructure. However, biosynthesis of FAEEs is limited by the supply of precursor lipids and acetyl-CoA. In the present study, we explored the production potential of an engineered biosynthetic pathway coupled to the addition of ethanol in the oleaginous yeast Yarrowia lipolytica...
April 25, 2018: ACS Synthetic Biology
Xiangfeng Huang, Huijuan Luo, Tianshuai Mu, Yi Shen, Ming Yuan, Jia Liu
Low concentrations of acetic acid were used as carbon source to cultivate Cryptococcus curvatus MUCL 29819 for lipid production under high content of ammonia. Phosphorus limitation combined with initial pH regulation (pH = 6) weakened inhibition of free ammonia and promoted lipid accumulation. In batch cultivation, the produced lipid content and yield was 30.3% and 0.92 g/L, higher than those under unlimited condition (18.3% and 0.64 g/L). The content of monounsaturated fatty acid also increased from 37...
April 18, 2018: Bioresource Technology
Batul Diwan, Piyush Parkhey, Pratima Gupta
The reserves of fossil-based fuels, which currently seem sufficient to meet the global demands, is inevitably on the verge of exhaustion. Contemporary raw material for alternate fuel like biodiesel is usually edible plant commodity oils, whose increasing public consumption rate raises the need of finding a non-edible and fungible alternate oil source. In this quest, single cell oils (SCO) from oleaginous yeasts and fungi can provide a sustainable alternate of not only functional but also valuable (polyunsaturated fatty acids (PUFA)-rich) lipids...
April 23, 2018: Folia Microbiologica
Jayita Chopra, Ramkrishna Sen
Lipid accumulation in oleaginous yeast is generally induced by nitrogen starvation, while oxygen saturation can influence biomass growth. Systematic shake flask studies that help in identifying the right nitrogen source and relate its uptake kinetics to lipid biosynthesis under varying oxygen saturation conditions are very essential for addressing the bioprocessing-related issues, which are envisaged to occur in the fermenter scale production. In the present study, lipid bioaccumulation by P. guilliermondii at varying C:N ratios and oxygen transfer conditions (assessed in terms of kL a) was investigated in shake flasks using a pre-optimized N-source and a two-stage inoculum formulated in a hybrid medium...
April 20, 2018: Bioprocess and Biosystems Engineering
Neha Arora, Philip T Pienkos, Vikas Pruthi, Krishna Mohan Poluri, Michael T Guarnieri
Ongoing global efforts to commercialize microalgal biofuels have expedited the use of multi-omics techniques to gain insights into lipid biosynthetic pathways. Functional genomics analyses have recently been employed to complement existing sequence-level omics studies, shedding light on the dynamics of lipid synthesis and its interplay with other cellular metabolic pathways, thus revealing possible targets for metabolic engineering. Here, we review the current status of algal omics studies to reveal potential targets to augment TAG accumulation in various microalgae...
April 17, 2018: Biotechnology Advances
Jin-Lai Zhang, Yang-Zi Peng, Duo Liu, Hong Liu, Ying-Xiu Cao, Bing-Zhi Li, Chun Li, Ying-Jin Yuan
BACKGROUND: The oleaginous yeast Yarrowia lipolytica is a promising microbial cell factory due to their biochemical characteristics and native capacity to accumulate lipid-based chemicals. To create heterogenous biosynthesis pathway and manipulate metabolic flux in Y. lipolytica, numerous studies have been done for developing synthetic biology tools for gene regulation. CRISPR interference (CRISPRi), as an emerging technology, has been applied for specifically repressing genes of interest...
April 20, 2018: Microbial Cell Factories
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