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      Enhancing oil production and harvest by combining the marine alga Nannochloropsis oceanica and the oleaginous fungus Mortierella elongata

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          Abstract

          Background

          Although microalgal biofuels have potential advantages over conventional fossil fuels, high production costs limit their application in the market. We developed bio-flocculation and incubation methods for the marine alga, Nannochloropsis oceanica CCMP1779, and the oleaginous fungus, Mortierella elongata AG77, resulting in increased oil productivity.

          Results

          By growing separately and then combining the cells, the M. elongata mycelium could efficiently capture N. oceanica due to an intricate cellular interaction between the two species leading to bio-flocculation. Use of a high-salt culture medium induced accumulation of triacylglycerol (TAG) and enhanced the contents of polyunsaturated fatty acids (PUFAs) including arachidonic acid and docosahexaenoic acid in M. elongata. To increase TAG productivity in the alga, we developed an effective, reduced nitrogen-supply regime based on ammonium in environmental photobioreactors. Under optimized conditions, N. oceanica produced high levels of TAG that could be indirectly monitored by following chlorophyll content. Combining N. oceanica and M. elongata to initiate bio-flocculation yielded high levels of TAG and total fatty acids, with ~ 15 and 22% of total dry weight (DW), respectively, as well as high levels of PUFAs. Genetic engineering of N. oceanica for higher TAG content in nutrient-replete medium was accomplished by overexpressing DGTT5, a gene encoding the type II acyl-CoA:diacylglycerol acyltransferase 5. Combined with bio-flocculation, this approach led to increased production of TAG under nutrient-replete conditions (~ 10% of DW) compared to the wild type (~ 6% of DW).

          Conclusions

          The combined use of M. elongata and N. oceanica with available genomes and genetic engineering tools for both species opens up new avenues to improve biofuel productivity and allows for the engineering of polyunsaturated fatty acids.

          Electronic supplementary material

          The online version of this article (10.1186/s13068-018-1172-2) contains supplementary material, which is available to authorized users.

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          Most cited references56

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          Microalgae for oil: strain selection, induction of lipid synthesis and outdoor mass cultivation in a low-cost photobioreactor.

          Thirty microalgal strains were screened in the laboratory for their biomass productivity and lipid content. Four strains (two marine and two freshwater), selected because robust, highly productive and with a relatively high lipid content, were cultivated under nitrogen deprivation in 0.6-L bubbled tubes. Only the two marine microalgae accumulated lipid under such conditions. One of them, the eustigmatophyte Nannochloropsis sp. F&M-M24, which attained 60% lipid content after nitrogen starvation, was grown in a 20-L Flat Alveolar Panel photobioreactor to study the influence of irradiance and nutrient (nitrogen or phosphorus) deprivation on fatty acid accumulation. Fatty acid content increased with high irradiances (up to 32.5% of dry biomass) and following both nitrogen and phosphorus deprivation (up to about 50%). To evaluate its lipid production potential under natural sunlight, the strain was grown outdoors in 110-L Green Wall Panel photobioreactors under nutrient sufficient and deficient conditions. Lipid productivity increased from 117 mg/L/day in nutrient sufficient media (with an average biomass productivity of 0.36 g/L/day and 32% lipid content) to 204 mg/L/day (with an average biomass productivity of 0.30 g/L/day and more than 60% final lipid content) in nitrogen deprived media. In a two-phase cultivation process (a nutrient sufficient phase to produce the inoculum followed by a nitrogen deprived phase to boost lipid synthesis) the oil production potential could be projected to be more than 90 kg per hectare per day. This is the first report of an increase of both lipid content and areal lipid productivity attained through nutrient deprivation in an outdoor algal culture. The experiments showed that this marine eustigmatophyte has the potential for an annual production of 20 tons of lipid per hectare in the Mediterranean climate and of more than 30 tons of lipid per hectare in sunny tropical areas.
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            Exploiting diversity and synthetic biology for the production of algal biofuels.

            Modern life is intimately linked to the availability of fossil fuels, which continue to meet the world's growing energy needs even though their use drives climate change, exhausts finite reserves and contributes to global political strife. Biofuels made from renewable resources could be a more sustainable alternative, particularly if sourced from organisms, such as algae, that can be farmed without using valuable arable land. Strain development and process engineering are needed to make algal biofuels practical and economically viable.
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              Biodiesel production by microalgal biotechnology

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                Author and article information

                Contributors
                1-517-355-1609 , benning@msu.edu
                Journal
                Biotechnol Biofuels
                Biotechnol Biofuels
                Biotechnology for Biofuels
                BioMed Central (London )
                1754-6834
                22 June 2018
                22 June 2018
                2018
                : 11
                : 174
                Affiliations
                [1 ]ISNI 0000 0001 2150 1785, GRID grid.17088.36, Department of Energy-Plant Research Laboratory, , Michigan State University, ; East Lansing, MI 48824 USA
                [2 ]ISNI 0000 0001 2150 1785, GRID grid.17088.36, Department of Biochemistry and Molecular Biology, , Michigan State University, ; East Lansing, MI 48824 USA
                [3 ]ISNI 0000 0001 2364 4210, GRID grid.7450.6, Department of Plant Biochemistry, Albrecht-von-Haller-Institute for Plant Sciences, , Georg-August-University, ; 37073 Goettingen, Germany
                [4 ]ISNI 0000 0001 2150 1785, GRID grid.17088.36, Department of Plant, Soil and Microbial Sciences, , Michigan State University, ; East Lansing, MI 48824 USA
                [5 ]ISNI 0000 0001 2150 1785, GRID grid.17088.36, Great Lakes Bioenergy Research Center, , Michigan State University, ; East Lansing, MI 48824 USA
                [6 ]ISNI 0000 0001 2150 1785, GRID grid.17088.36, Department of Plant Biology, , Michigan State University, ; East Lansing, MI 48824 USA
                Author information
                http://orcid.org/0000-0001-8585-3667
                Article
                1172
                10.1186/s13068-018-1172-2
                6013958
                29977335
                cd8a7c77-9b65-4da2-b532-6ec986c423dd
                © The Author(s) 2018

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 16 March 2018
                : 12 June 2018
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100006132, Office of Science;
                Award ID: DE-FG02-91ER20021
                Award ID: DE-FC02-07ER64494
                Award ID: DE-SC0018409
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100000001, National Science Foundation;
                Award ID: DEB 1737898
                Award ID: NSF DBI-1358474
                Award Recipient :
                Categories
                Research
                Custom metadata
                © The Author(s) 2018

                Biotechnology
                nannochloropsis,mortierella,bio-flocculation,polyunsaturated fatty acid,triacylglycerol,photobioreactor,microalgae,filamentous fungi,cell–wall interaction,biofuel,nitrogen starvation

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