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Journal articles on the topic 'Thraustochytrids'

Author: Grafiati

Published: 9 September 2021

Last updated: 12 February 2022

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1

Hà, Nguyễn Thị Hoài, Phạm Thị Bích Đào, and Nguyễn Đình Tuấn. "Taxonomic characterization of ten Thraustochytrids strains isolated from mangrove Xuan Thuy, Nam Dinh." Vietnam Journal of Biotechnology 14, no. 2 (June 30, 2016): 377–84. http://dx.doi.org/10.15625/1811-4989/14/2/9364.

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Thraustochytrids have become of considerable industrial and scientific interest in the past decade due to their health benefits. Thraustochytrids are found in a wide variety of marine habitats such as the coastal, mangrove and sediments including the deep sea. Thraustochytrids are extremely common on the detritus, macroalgae and decaying leaf, they play an important role as organic matter-degrading microorganisms Thraustochytrids are unicellular, eukaryotic, chemo-organotrophic organisms. Ten thraustochytrids strains PT269, PT270, PT273, PT274, PT279, PT284, PT285, PT287, PT81, PT84 were isolated from four locations in Xuan Thuy mangroves, Nam Dinh. In this report, classification is based on morphology and 18S rDNA sequences. Ten Thraustochytrid strains could be classified into three types of colony and four types of cell morphology. Molecular phylogenetic analysis of 18S rDNA sequences showed homology score to be 99-100% and these strains belonged to four genera in the family Thraustochytriaceae. PT269, PT279, PT284 and PT287 strains belong to Aurantiochytrium genus, they produce amoeboid cells and occur successive binary division. PT273 and PT285 strains belong to Thraustochytrium genus, thallus directly develop and cleave into sporangium. PT274 strain belong to Aplanochytrium genus with two distinct development, amoeboid cells are found, they rapidly round up and become sporangium; and successive binary cell division. PT270, PT81 and PT84 strains belong to genus Schizochytrium, they have successive binary cell division, zoospores release.

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2

Kobayashi, Takumi, Keishi Sakaguchi, Takanori Matsuda, Eriko Abe, Yoichiro Hama, Masahiro Hayashi, Daiske Honda, et al. "Increase of Eicosapentaenoic Acid in Thraustochytrids through Thraustochytrid Ubiquitin Promoter-Driven Expression of a Fatty Acid Δ5 Desaturase Gene." Applied and Environmental Microbiology 77, no. 11 (April 8, 2011): 3870–76. http://dx.doi.org/10.1128/aem.02664-10.

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ABSTRACTThraustochytrids, marine protists known to accumulate polyunsaturated fatty acids (PUFAs) in lipid droplets, are considered an alternative to fish oils as a source of PUFAs. The major fatty acids produced in thraustochytrids are palmitic acid (C16:0),n− 6 docosapentaenoic acid (DPA) (C22:5n− 6), and docosahexaenoic acid (DHA) (C22:6n− 3), with eicosapentaenoic acid (EPA) (C20:5n− 3) and arachidonic acid (AA) (C20:4n− 6) as minor constituents. We attempted here to alter the fatty acid composition of thraustochytrids through the expression of a fatty acid Δ5 desaturase gene driven by the thraustochytrid ubiquitin promoter. The gene was functionally expressed inAurantiochytrium limacinummh0186, increasing the amount of EPA converted from eicosatetraenoic acid (ETA) (C20:4n− 3) by the Δ5 desaturase. The levels of EPA and AA were also increased by 4.6- and 13.2-fold in the transgenic thraustochytrids compared to levels in the mock transfectants when ETA and dihomo-γ-linolenic acid (DGLA) (C20:3n− 6) were added to the culture at 0.1 mM. Interestingly, the amount of EPA in the transgenic thraustochytrids increased in proportion to the amount of ETA added to the culture up to 0.4 mM. The rates of conversion and accumulation of EPA were much higher in the thraustochytrids than in baker's yeasts when the desaturase gene was expressed with the respective promoters. This report describes for the first time the finding that an increase of EPA could be accomplished by introducing the Δ5 desaturase gene into thraustochytrids and indicates that molecular breeding of thraustochytrids is a promising strategy for generating beneficial PUFAs.

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Rau, E.-Ming, and Helga Ertesvåg. "Method Development Progress in Genetic Engineering of Thraustochytrids." Marine Drugs 19, no. 9 (September 11, 2021): 515. http://dx.doi.org/10.3390/md19090515.

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Thraustochytrids are unicellular, heterotrophic marine eukaryotes. Some species are known to store surplus carbon as intracellular lipids, and these also contain the long-chain polyunsaturated fatty acid docosahexaenoic acid (DHA). Most vertebrates are unable to synthesize sufficient amounts of DHA, and this fatty acid is essential for, e.g., marine fish, domesticated animals, and humans. Thraustochytrids may also produce other commercially valuable fatty acids and isoprenoids. Due to the great potential of thraustochytrids as producers of DHA and other lipid-related molecules, a need for more knowledge on this group of organisms is needed. This necessitates the ability to do genetic manipulation of the different strains. Thus far, this has been obtained for a few strains, while it has failed for other strains. Here, we systematically review the genetic transformation methods used for different thraustochytrid strains, with the aim of aiding studies on strains not yet successfully transformed. The designs of transformation cassettes are also described and compared. Moreover, the potential problems when trying to establish transformation protocols in new thraustochytrid species/strains are discussed, along with suggestions utilized in other organisms to overcome similar challenges. The approaches discussed in this review could be a starting point when designing protocols for other non-model organisms.

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4

Sakaguchi, Keishi, Takanori Matsuda, Takumi Kobayashi, Jun-ichiro Ohara, Rie Hamaguchi, Eriko Abe, Naoki Nagano, et al. "Versatile Transformation System That Is Applicable to both Multiple Transgene Expression and Gene Targeting for Thraustochytrids." Applied and Environmental Microbiology 78, no. 9 (February 17, 2012): 3193–202. http://dx.doi.org/10.1128/aem.07129-11.

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ABSTRACTA versatile transformation system for thraustochytrids, a promising producer for polyunsaturated fatty acids and fatty acid-derived fuels, was established. G418, hygromycin B, blasticidin, and zeocin inhibited the growth of thraustochytrids, indicating that multiple selectable marker genes could be used in the transformation system. A neomycin resistance gene (neor), driven with an ubiquitin or an EF-1α promoter-terminator fromThraustochytrium aureumATCC 34304, was introduced into representatives of two thraustochytrid genera,AurantiochytriumandThraustochytrium. Theneormarker was integrated into the chromosomal DNA by random recombination and then functionally translated intoneormRNA. Additionally, we confirmed that another two genera,ParietichytriumandSchizochytrium, could be transformed by the same method. By this method, the enhanced green fluorescent protein was functionally expressed in thraustochytrids. Meanwhile,T. aureumATCC 34304 could be transformed by two 18S ribosomal DNA-targeting vectors, designed to cause single- or double-crossover homologous recombination. Finally, the fatty acid Δ5 desaturase gene was disrupted by double-crossover homologous recombination inT. aureumATCC 34304, resulting in an increase of dihomo-γ-linolenic acid (C20:3n-6) and eicosatetraenoic acid (C20:4n-3), substrates for Δ5 desaturase, and a decrease of arachidonic acid (C20:4n-6) and eicosapentaenoic acid (C20:5n-3), products for the enzyme. These results clearly indicate that a versatile transformation system which could be applicable to both multiple transgene expression and gene targeting was established for thraustochytrids.

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5

Đào, Phạm Thị Bích, Nguyễn Đình Tuấn, Trần Đăng Khoa, Chử Thị Huyên, Đỗ Hoàng Thành, and Nguyễn Thị Hoài Hà. "Lipid biosynthesis of ten thraustochytrid strains isolated from mangrove Xuan Thuy, Nam Dinh." Vietnam Journal of Biotechnology 14, no. 2 (June 30, 2016): 385–92. http://dx.doi.org/10.15625/1811-4989/14/2/9365.

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The features of polyunsaturated fatty acid-PUFA structures were corresponded to each separate functions as adjusting the cellular physiology and gene expression. Therefore, lack of PUFA could lead to abnormalities in skin, kidney, neural networks, immune responses and inflammation; cardiovascular, endocrine, respiratory and reproductive systems. In fish oil, PUFA content were low, thus it was difficult to produce on a large scale. Therefore, the exploration of PUFA sources particularly as arachidonic acid-AA, eicosapentaenoic acid EPA, docosapentaenoic acid-DPA/DHA attracted many researches. Heterotrophic microalgae Thraustochytrids were capable of producing high amounts of DHA and PUFA composition varied. DHA can be synthesized by the metabolism of AA, EPA and DPA. The different types of PUFA reflected relationships in classification. Ten heterotrophic microalgae thraustochytrids isolated from mangrove Xuan Thuy, Nam Dinh contain fatty acid composition varied from C12 to C28. Especially, they had two important fatty acids of PUFA as EPA and DPA. Polyunsaturated fatty acids - PUFA content of ten thraustochytrid strains were from 28.95 to 49.62% total lipid. DPA compared to other PUFA were high for all thraustochytrid strains studied, accounting 20.22 to 39.35% TFA. Ten thraustochytrid strains had the highest growth with carbon source as glucose, total lipid reached 7 to 12.35 % dry weight biomass after 72 hours. Growth rate and lipid biosynthesis in organic nitrogen source were higher than in inorganic nitrogen sources. The best source of nitrogen for growth and lipid biosynthesis of ten thraustochytrid strains is yeast extract, total lipid were 8.57 to 18.87% dry weight biomass after 72 hours.

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6

Bartosova, Zdenka, Helga Ertesvåg, Eirin Lishaugen Nyfløt, Kristoffer Kämpe, Inga Marie Aasen, and Per Bruheim. "Combined Metabolome and Lipidome Analyses for In-Depth Characterization of Lipid Accumulation in the DHA Producing Aurantiochytrium sp. T66." Metabolites 11, no. 3 (February 25, 2021): 135. http://dx.doi.org/10.3390/metabo11030135.

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Thraustochytrids are marine heterotrophic microorganisms known for their potential to accumulate docosahexaenoic acid (DHA)-enriched lipids. There have been many attempts to improve thraustochytrid DHA bioprocesses, especially through traditional optimization of cultivation and media conditions. Nevertheless, thraustochytrid-based bioprocesses are still not commercially competitive for high volume-low cost production of DHA. Thus, it is realized that genetic and metabolic engineering strategies are needed for the development of commercially competitive thraustochytrid DHA cell factories. Here, we present an analytical workflow for high resolution phenotyping at metabolite and lipid levels to generate deeper insight into the thraustochytrid physiology, with particular focus on central carbon and redox metabolism. We use time-series sampling during unlimited growth and nitrogen depleted triggering of DHA synthesis and lipid accumulation (LA) to show-case our methodology. The mass spectrometric absolute quantitative metabolite profiling covered glycolytic, pentose phosphate pathway (PPP) and tricarboxylic acid cycle (TCA) metabolites, amino acids, complete (deoxy)nucleoside phosphate pools, CoA and NAD metabolites, while semiquantitative high-resolution supercritical fluid chromatography MS/MS was applied for the lipid profiling. Interestingly, trace amounts of a triacylglycerols (TG) with DHA incorporated in all three acyl positions was detected, while TGs 16:0_16:0_22:6 and 16:0_22:6_22:6 were among the dominant lipid species. The metabolite profiling data indicated that lipid accumulation is not limited by availability of the acyl chain carbon precursor acetyl-CoA nor reducing power (NADPH) but rather points to the TG head group precursor glycerol-3-phosphate as the potential cause at the metabolite level for the gradual decline in lipid production throughout the cultivation. This high-resolution phenotyping provides new knowledge of changes in the central metabolism during growth and LA in thraustochytrids and will guide target selection for metabolic engineering needed for further improvements of this DHA cell factory.

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Stefánsson, Magnús Örn, Sigurður Baldursson, Kristinn P. Magnússon, Arnheiður Eyþórsdóttir, and Hjörleifur Einarsson. "Isolation, Characterization and Biotechnological Potentials of Thraustochytrids from Icelandic Waters." Marine Drugs 17, no. 8 (July 31, 2019): 449. http://dx.doi.org/10.3390/md17080449.

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The following study reports on the first thraustochytrid isolates identified from Iceland. They were collected from three different locations off the northern coast of the country (Location A, Skagaströnd; Location B, Hveravík; and Location C, Eyjafjörður). Using 18S rDNA sequence analysis, isolates from Locations A and B were identified within the Thraustochytrium kinnei species while other isolates within the Sicyoidochytrium minutum species when compared to other known strains. Cells isolated from Locations A ( 2 . 10 ± 0 . 70 g/L) and B ( 1 . 54 ± 0 . 17 g/L) produced more biomass than the ones isolated from Location C ( 0 . 43 ± 0 . 02 g/L). This study offers the first-time examination of the utility of byproducts from fisheries as a nitrogen source in media formulation for thraustochytrids. Experiments showed that isolates produced more biomass (per unit of substrate) when cultured on nitrogen of marine ( 2 . 55 ± 0 . 74 g/L) as compared to of commercial origin ( 1 . 06 ± 0 . 57 g/L). Glycerol ( 2 . 43 ± 0 . 56 g/L) was a better carbon source than glucose ( 1 . 84 ± 0 . 57 g/L) in growth studies. Fatty acid (FA) profiles showed that the isolates from Location C (S. minutum) had low ratios of monounsaturated ( 4 . 21 ± 2 . 96 % ) and omega-6 ( 0 . 68 ± 0 . 59 % ) FAs. However, the isolates also had high ratios of docosahexaenoic acid (DHA; 35 . 65 ± 1 . 73 % ) and total omega-3 FAs ( 40 . 39 ± 2 . 39 % ), indicating that they could serve as a source of marine oils for human consumption and in aquaculture feeds. The T. kinnei isolates from Location A could be used in biodiesel production due to their high ratios of monounsaturated ( 18 . 38 ± 6 . 27 % ) long chain ( 57 . 43 ± 8 . 27 % ) FAs.

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8

Rau, E.-Ming, Inga Marie Aasen, and Helga Ertesvåg. "A non-canonical Δ9-desaturase synthesizing palmitoleic acid identified in the thraustochytrid Aurantiochytrium sp. T66." Applied Microbiology and Biotechnology 105, no. 14-15 (July 22, 2021): 5931–41. http://dx.doi.org/10.1007/s00253-021-11425-5.

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Abstract Thraustochytrids are oleaginous marine eukaryotic microbes currently used to produce the essential omega-3 fatty acid docosahexaenoic acid (DHA, C22:6 n-3). To improve the production of this essential fatty acid by strain engineering, it is important to deeply understand how thraustochytrids synthesize fatty acids. While DHA is synthesized by a dedicated enzyme complex, other fatty acids are probably synthesized by the fatty acid synthase, followed by desaturases and elongases. Which unsaturated fatty acids are produced differs between different thraustochytrid genera and species; for example, Aurantiochytrium sp. T66, but not Aurantiochytrium limacinum SR21, synthesizes palmitoleic acid (C16:1 n-7) and vaccenic acid (C18:1 n-7). How strain T66 can produce these fatty acids has not been known, because BLAST analyses suggest that strain T66 does not encode any Δ9-desaturase-like enzyme. However, it does encode one Δ12-desaturase-like enzyme. In this study, the latter enzyme was expressed in A. limacinum SR21, and both C16:1 n-7 and C18:1 n-7 could be detected in the transgenic cells. Our results show that this desaturase, annotated T66Des9, is a Δ9-desaturase accepting C16:0 as a substrate. Phylogenetic studies indicate that the corresponding gene probably has evolved from a Δ12-desaturase-encoding gene. This possibility has not been reported earlier and is important to consider when one tries to deduce the potential a given organism has for producing unsaturated fatty acids based on its genome sequence alone. Key points • In thraustochytrids, automatic gene annotation does not always explain the fatty acids produced. • T66Des9 is shown to synthesize palmitoleic acid (C16:1 n-7). • T66des9 has probably evolved from Δ12-desaturase-encoding genes.

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Leyland, Ben, Stefan Leu, and Sammy Boussiba. "Are Thraustochytrids algae?" Fungal Biology 121, no. 10 (October 2017): 835–40. http://dx.doi.org/10.1016/j.funbio.2017.07.006.

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10

Leyton, Allison, Liset Flores, Carolina Shene, Yusuf Chisti, Giovanni Larama, Juan A. Asenjo, and Roberto E. Armenta. "Antarctic Thraustochytrids as Sources of Carotenoids and High-Value Fatty Acids." Marine Drugs 19, no. 7 (July 6, 2021): 386. http://dx.doi.org/10.3390/md19070386.

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Eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and carotenoids are needed as human dietary supplements and are essential components in commercial feeds for the production of aquacultured seafood. Microorganisms such as thraustochytrids are potential natural sources of these compounds. This research reports on the lipid and carotenoid production capacity of thraustochytrids that were isolated from coastal waters of Antarctica. Of the 22 isolates, 21 produced lipids containing EPA+DHA, and the amount of these fatty acids exceeded 20% of the total fatty acids in 12 isolates. Ten isolates were shown to produce carotenoids (27.4–63.9 μg/g dry biomass). The isolate RT2316-16, identified as Thraustochytrium sp., was the best producer of biomass (7.2 g/L in five days) rich in carotenoids (63.9 μg/g) and, therefore, became the focus of this investigation. The main carotenoids in RT2316-16 were β-carotene and canthaxanthin. The content of EPA+DHA in the total lipids (34 ± 3% w/w in dry biomass) depended on the stage of growth of RT2316-16. Lipid and carotenoid content of the biomass and its concentration could be enhanced by modifying the composition of the culture medium. The estimated genome size of RT2316-16 was 44 Mb. Of the 5656 genes predicted from the genome, 4559 were annotated. These included genes of most of the enzymes in the elongation and desaturation pathway of synthesis of ω-3 polyunsaturated fatty acids. Carotenoid precursors in RT2316-16 were synthesized through the mevalonate pathway. A β-carotene synthase gene, with a different domain organization compared to the gene in other thraustochytrids, explained the carotenoid profile of RT2316-16.

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MALAWET, Tarnhatai, Phuwadol BANGRAK, Yuwadee PEERAPORNPISAL, and Niyom KAMLANGDEE. "Newly Isolated High Squalene Producing Thraustochytrid Strain Aurantochytrium sp. P5/2 from Mangrove Habitats in Nakhon Si Thammarat Province, Thailand." Walailak Journal of Science and Technology (WJST) 17, no. 3 (June 20, 2019): 212–21. http://dx.doi.org/10.48048/wjst.2020.6476.

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Thraustochytrids are alternative potential sources of squalene, because they grow rapidly, are relatively easy to culture, and accumulate in large amounts. The objectives of this research were to isolate squalene-producing Thraustochytrids from fallen leaves in Paknakon Bay, including Paknakon Mangrove forest (N), Pakpanang Mangrove forest (P) and Thasala Mangrove forest (T), Nakhon Si Thammarat, Thailand, and to investigate their total lipid profile and squalene contents. A total of nine Thraustochytrid isolates were obtained.Morphological and molecular features revealed that those Thraustochytrids belonged to the genus Aurantiochytrium (N1, N14, P1/1, P5/2, P6/1, P43, T1, T26, and T42). Subsequently, they were cultivated and their cell dry weight, fatty acid compositions, and squalene contents were analyzed. At 96 h of cultivation, the dry cell weights ranged from 7.51 to 17.43 mg/g. The total lipid profile showed a broad spectrum of saturated fatty acids with an abundance of palmitic acid (16:0), 24.72 - 41.06 % TFA, pentadecanoic acid (15:0) 16.75 - 28.48 % TFA, heptadecanoic acid (17:0) 4.19 - 7.67 % TFA, lignoceric acid (24:0) 2.76 - 8.83 % TFA, myristic acid (14:0) 2.17 - 3.43 % TFA, stearic acid (18:0) 0.83 - 1.32 % TFA, arachidic acid (20:0) 0.19 - 0.33 % TFA, and behenic acid (22:0) 0.19 - 0.21 % TFA, respectively. Unsaturated fatty acids, including Docosahexaaenoic acid (22:6; 8.59 - 35.99 % TFA), Clupanodonic acid (22:5, 2.24 - 8.94 % TFA), Arachidonic acid (20:4, 0.32 - 0.60 % TFA), Eicosapentaenoic acid (20:5, 0.19 - 0.62 % TFA), Linolenic acid (18:3, 0.12 - 0.18 % TFA), and Erucic acid (22:1; 0.02 - 0.09 % TFA) were also found. The squalene contents ranged from 0.06 to 4.78 mg/g. The highest biomass and squalene-accumulation was achieved from strain P5/2, which was identified as Aurantiochytrium sp.ม with a maximum yield of 4.78 mg/g at 96 h of cultivation.

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Nham Tran, Thi Linh, Ana F. Miranda, Aidyn Mouradov, and Benu Adhikari. "Physicochemical Characteristics of Protein Isolated from Thraustochytrid Oilcake." Foods 9, no. 6 (June 11, 2020): 779. http://dx.doi.org/10.3390/foods9060779.

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The oil from thraustochytrids, unicellular heterotrophic marine protists, is increasingly used in the food and biotechnological industries as it is rich in omega-3 fatty acids, squalene and a broad spectrum of carotenoids. This study showed that the oilcake, a by-product of oil extraction, is equally valuable as it contained 38% protein/dry mass, and thraustochytrid protein isolate can be obtained with 92% protein content and recovered with 70% efficiency. The highest and lowest solubilities of proteins were observed at pH 12.0 and 4.0, respectively, the latter being its isoelectric point. Aspartic acid, glutamic acid, histidine, and arginine were the most abundant amino acids in proteins. The arginine-to-lysine ratio was higher than one, which is desired in heart-healthy foods. The denaturation temperature of proteins ranged from 167.8–174.5 °C, indicating its high thermal stability. Proteins also showed high emulsion activity (784.1 m2/g) and emulsion stability (209.9 min) indices. The extracted omega-3-rich oil melted in the range of 30–34.6 °C and remained stable up to 163–213 °C. This study shows that thraustochytrids are not only a valuable source of omega 3-, squalene- and carotenoid-containing oils, but are also rich in high-value protein with characteristics similar to those from oilseeds.

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Jakobsen, Anita N., Inga M. Aasen, and Arne R. Strøm. "Endogenously Synthesized (−)-proto-Quercitol and Glycine Betaine Are Principal Compatible Solutes of Schizochytrium sp. Strain S8 (ATCC 20889) and Three New Isolates of Phylogenetically Related Thraustochytrids." Applied and Environmental Microbiology 73, no. 18 (July 27, 2007): 5848–56. http://dx.doi.org/10.1128/aem.00610-07.

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ABSTRACT We report that endogenously synthesized (−)-proto-quercitol (1d-1,3,4/2,5-cyclohexanepentol) and glycine betaine were the principal compatible solutes of Schizochytrium sp. strain S8 (ATCC 20889) and three new osmotolerant isolates of thraustochytrids (strains T65, T66, and T67). The compatible solutes were identified and quantified by use of nuclear magnetic resonance spectroscopy, and their identity was confirmed by mass spectroscopy and measurement of the specific optical rotation. The cellular content of compatible solutes increased with increasing NaCl concentration of a defined medium. (−)-proto-Quercitol was the dominating solute at all NaCl concentrations tested (0.25 to 1.0 M), e.g., cells of S8 and T66 stressed with 1.0 M NaCl accumulated about 500 μmol (−)-proto-quercitol and 100 μmol glycine betaine per g dry weight. To our knowledge, (−)-proto-quercitol has previously been found only in eucalyptus. The 18S rRNA gene sequences of the four (−)-proto-quercitol-producing strains showed 99% identity, and they displayed the same fatty acid profile. The only polyunsaturated fatty acids accumulated were docosahexaenoic acid (78%) and docosapentaenoic acid (22%). A less osmotolerant isolate (strain T29), which was closely phylogenetically related to Thraustochytrium aureum (ATCC 34304), did not contain (−)-proto-quercitol or glycine betaine. Thus, the level of osmotolerance and the osmolyte systems vary among thraustochytrids.

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Lee-Chang, Kim Jye, Matthew C. Taylor, Guy Drummond, Roger J. Mulder, Maged Peter Mansour, Mina Brock, and Peter D. Nichols. "Docosahexaenoic Acid Is Naturally Concentrated at the sn-2 Position in Triacylglycerols of the Australian Thraustochytrid Aurantiochytrium sp. Strain TC 20." Marine Drugs 19, no. 7 (July 1, 2021): 382. http://dx.doi.org/10.3390/md19070382.

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The Labyrinthulomycetes or Labyrinthulea are a class of protists that produce a network of filaments that enable the cells to glide along and absorb nutrients. One of the main two Labyrinthulea groups is the thraustochytrids, which are becoming an increasingly recognised and commercially used alternate source of long-chain (LC, ≥C20) omega-3 containing oils. This study demonstrates, to our knowledge for the first time, the regiospecificity of the triacylglycerol (TAG) fraction derived from Australian thraustochytrid Aurantiochytrium sp. strain TC 20 obtained using 13C nuclear magnetic resonance spectroscopy (13C NMR) analysis. The DHA present in the TC 20 TAG fraction was determined to be concentrated in the sn-2 position, with TAG (16:0/22:6/16:0) identified as the main species present. The sn-2 preference is similar to that found in salmon and tuna oil, and differs to seal oil containing largely sn-1,3 LC-PUFA. A higher concentration of sn-2 DHA occurred in the thraustochytrid TC 20 oil compared to that of tuna oil.

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Sen, Biswarup, Jiaqian Li, Lyu Lu, Mohan Bai, Yaodong He, and Guangyi Wang. "Elemental Composition and Cell Mass Quantification of Cultured Thraustochytrids Unveil Their Large Contribution to Marine Carbon Pool." Marine Drugs 19, no. 9 (August 29, 2021): 493. http://dx.doi.org/10.3390/md19090493.

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The element stoichiometry of bacteria has received considerable attention because of their significant role in marine ecosystems. However, relatively little is known about the composition of major structural elements of the unicellular heterotrophic protists—thraustochytrids, despite their widely recognized contribution to marine nutrient cycling. Here, we analyze the cell volume and elemental C, N, H, and S cell content of seven cultured thraustochytrids, isolated from different marine habitats, in the exponential and stationary growth phases. We further derive the relationships between the cell volume and elemental C and N content of the cultured thraustochytrids. The cell volumes varied significantly (p < 0.001) among the isolates, with median values of 96.9 and 212.5 μm3 in the exponential and stationary phases, respectively. Our results showed a significantly higher percentage of C (64.0 to 67.5) and H (9.9 to 13.2) but a lower percentage of N (1.86 to 2.16) and S (0.34 to 0.91) in the stationary phase, along with marked variations of C and N fractions among isolates in the exponential phase. The cell C (5.7 to 203.7 pg) and N (0.65 to 6.1 pg) content exhibited a significant (p < 0.001) linear relationship with the cell volume (27.7 to 510 μm3). On further analysis of the relationship across the two growth phases, we found the equation (cell C (pg) = 0.356 × cell volume (μm3) + 20.922) for stationary phase cells more appropriate for C estimation of natural thraustochytrids. This study provides the first experimental evidence of higher cell C density than the current estimate and relatively larger C contribution of thraustochytrids than bacteria to the marine organic pool.

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Morabito, Christian, Caroline Bournaud, Cécile Maës, Martin Schuler, Riccardo Aiese Cigliano, Younès Dellero, Eric Maréchal, Alberto Amato, and Fabrice Rébeillé. "The lipid metabolism in thraustochytrids." Progress in Lipid Research 76 (October 2019): 101007. http://dx.doi.org/10.1016/j.plipres.2019.101007.

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17

Lewis, Tom E., Peter D. Nichols, and Thomas A. McMeekin. "The Biotechnological Potential of Thraustochytrids." Marine Biotechnology 1, no. 6 (November 1999): 580–87. http://dx.doi.org/10.1007/pl00011813.

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Cox, Serena L., Debbie Hulston, and Elizabeth W. Maas. "Cryopreservation of marine thraustochytrids (Labyrinthulomycetes)." Cryobiology 59, no. 3 (December 2009): 363–65. http://dx.doi.org/10.1016/j.cryobiol.2009.09.001.

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Humhal, Tomáš, Olga Kronusová, Petr Kaštánek, Tomáš Potočár, Jana Kohoutková, and Tomáš Brányik. "Influence of nitrogen sources on growth of thraustochytrids in waste water from the demineralization of cheese whey." Czech Journal of Food Sciences 37, No. 5 (October 31, 2019): 383–90. http://dx.doi.org/10.17221/172/2018-cjfs.

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An experimental design was ued to optimize the growth of two thraustochytrids, (Schizochytrium limacinum PA-968 and Japonochytrium marinum AN-4), on different nitrogen sources (yeast extract, corn steep liquor, ammonium sulphate) supplemented into saline waste water from the demineralization of cheese whey. Yeast extract was found to be the most suitable complex nutrient source. Nitrogen limitation was found to increase the lipid content in shake flask cultures of thraustochytrids by 12.7–22.4% w/w. The maximum total lipid content (79.1% w/w) and docosahexaenoic acid productivity (0.465 g/l per day) were achieved by J. marinum AN-4 in shake flask cultures. Fed-batch cultures of J. marinum AN-4, under conditions of nitrogen limitation, yielded biomass with a lower lipid content (72.1% wt.) but higher docosahexaenoic acid productivity (1.43 g/l per day). These results provide proof of concept that fed-batch cultivation of thraustochytrids, combined with nitrogen limitation, can be an appropriate strategy for the productive use of saline waste water from the dairy industry.

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Patel, Alok, Ulrika Rova, Paul Christakopoulos, and Leonidas Matsakas. "Assessment of Fatty Acids Profile and Omega-3 Polyunsaturated Fatty Acid Production by the Oleaginous Marine Thraustochytrid Aurantiochytrium sp. T66 Cultivated on Volatile Fatty Acids." Biomolecules 10, no. 5 (April 29, 2020): 694. http://dx.doi.org/10.3390/biom10050694.

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Thraustochytrids are considered natural producers of omega-3 fatty acids as they can synthesize up to 70% docosahexaenoic acids (DHA) of total lipids. However, commercial and sustainable production of microbial DHA is limited by elevated cost of carbon substrates for thraustochytrids cultivation. This problem can be addressed by utilizing low-cost renewable substrates. In the present study, growth, lipid accumulation and fatty acid profiles of the marine thraustochytrid Aurantiochytrium sp. T66 (ATCC-PRA-276) cultivated on volatile fatty acids (C1, formic acid; C2, acetic acid; C3, propionic acid; C4, butyric acid; C5, valeric acid and C6, caproic acid) and glucose as control were evaluated for the first time. This strain showed an inability to utilize C3, C5 and C6 as a substrate when provided at >2 g/L, while efficiently utilizing C2 and C4 up to 40 g/L. The highest cell dry weight (12.35 g/L) and total lipid concentration (6.59 g/L) were attained when this strain was cultivated on 40 g/L of butyric acid, followed by cultivation on glucose (11.87 g/L and 5.34 g/L, respectively) and acetic acid (8.70 g/L and 3.43 g/L, respectively). With 40 g/L butyric acid, the maximum docosahexaenoic acid content was 2.81 g/L, corresponding to 42.63% w/w of total lipids and a yield of 0.23 g/gcell dry weight (CDW). This marine oleaginous microorganism showed an elevated potential for polyunsaturated fatty acids production at higher acetic and butyric acid concentrations than previously reported. Moreover, fluorescence microscopy revealed that growth on butyric acid caused cell size to increase to 45 µm, one of the largest values reported for oleaginous microorganisms, as well as the presence of numerous tiny lipid droplets.

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Naganuma, T., H. Takasugi, and H. Kimura. "Abundance of thraustochytrids in coastal plankton." Marine Ecology Progress Series 162 (1998): 105–10. http://dx.doi.org/10.3354/meps162105.

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Pang, Ka-Lai, Min-Chiau Chen, Michael W. L. Chiang, Yu-Fen Huang, Hui-Yin Yeong, and Siew-Moi Phang. "Cu(II) pollution affects fecundity of the mangrove degrader community, the Labyrinthulomycetes." Botanica Marina 58, no. 2 (April 1, 2015): 129–38. http://dx.doi.org/10.1515/bot-2015-0006.

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Abstract Thraustochytrids, a group of fungus-like organisms belonging to the protist class Labyrinthulomycetes, are common colonisers and degraders of fallen leaves in mangroves and, thus, are actively involved in nutrient cycling. Mangroves, often located at the river mouth, constantly receive freshwater runoff, which contains organic and inorganic pollutants, including metals. Metals are known to cause cellular damage, which may affect the survival of thraustochytrids in the mangrove environment. A previous study suggested that Cu(II), one of the major metal ions in coastal water and sediment, retards the growth of and causes cellular damage to mangrove thraustochytrids. We hypothesize that increased concentrations of Cu(II) negatively affect the fecundity of mangrove thraustochytrids. In a laboratory study, we assessed the sporulation success (number of zoospores produced per colony) and the growth response (biomass) of 11 isolates of Schizochytrium limacinum collected from four mangrove stands in Taiwan exposed to increasing concentrations of Cu(II). Tolerance to Cu(II) varied among the tested isolates of S. limacinum. In general, a negative dose-response relationship was exhibited between growth response/sporulation success and increasing concentrations of Cu(II). However, exposure to low concentrations of Cu(II) had a stimulating effect on growth (2 mg l-1) and sporulation (2–64 mg l-1) for some isolates. A sharp decline in growth was observed at 32 mg l-1 Cu(II), and sporulation success was more tolerant to increasing concentrations of Cu(II). The IC10 and IC50 values for growth were 1.0–16.5 mg l-1 and 9.1–23.9 mg l-1, respectively, whereas those for sporulation success were 0.6–40.7 mg l-1 and 10.5–108.4 mg l-1, respectively. In conclusion, Cu(II) interfered with both the growth and sporulation success of S. limacinum, which may affect its abundance and distribution in mangrove environments.

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Nham Tran, Thi Linh, Ana F. Miranda, Adarsha Gupta, Munish Puri, Andrew S. Ball, Benu Adhikari, and Aidyn Mouradov. "The Nutritional and Pharmacological Potential of New Australian Thraustochytrids Isolated from Mangrove Sediments." Marine Drugs 18, no. 3 (March 6, 2020): 151. http://dx.doi.org/10.3390/md18030151.

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Mangrove sediments represent unique microbial ecosystems that act as a buffer zone, biogeochemically recycling marine waste into nutrient-rich depositions for marine and terrestrial species. Marine unicellular protists, thraustochytrids, colonizing mangrove sediments have received attention due to their ability to produce large amounts of long-chain ω3-polyunsaturated fatty acids. This paper represents a comprehensive study of two new thraustochytrids for their production of valuable biomolecules in biomass, de-oiled cakes, supernatants, extracellular polysaccharide matrixes, and recovered oil bodies. Extracted lipids (up to 40% of DW) rich in polyunsaturated fatty acids (up to 80% of total fatty acids) were mainly represented by docosahexaenoic acid (75% of polyunsaturated fatty acids). Cells also showed accumulation of squalene (up to 13 mg/g DW) and carotenoids (up to 72 µg/g DW represented by astaxanthin, canthaxanthin, echinenone, and β-carotene). Both strains showed a high concentration of protein in biomass (29% DW) and supernatants (2.7 g/L) as part of extracellular polysaccharide matrixes. Alkalinization of collected biomass represents a new and easy way to recover lipid-rich oil bodies in the form of an aqueous emulsion. The ability to produce added-value molecules makes thraustochytrids an important alternative to microalgae and plants dominating in the food, pharmacological, nutraceutical, and cosmetics industries.

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Zhang, Yujie, Valerie Ward, Dorothy Dennis, Natalia Plechkova, Roberto Armenta, and Lars Rehmann. "Efficient Extraction of a Docosahexaenoic Acid (DHA)-Rich Lipid Fraction from Thraustochytrium sp. Using Ionic Liquids." Materials 11, no. 10 (October 15, 2018): 1986. http://dx.doi.org/10.3390/ma11101986.

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Polyunsaturated fatty acids (PUFAs) play a significant role in the modulation and prevention of various diseases, and hence are attracting increasing attention from the biotech industry. Thraustochytrids are marine heterokonts that exhibit robust growth rates, high PUFA content, and more specifically, a large percentage of omega-3 fatty acids like docosahexaenoic acid (DHA). Recently, ionic liquids (ILs) have been shown to improve the efficiency of organic solvent extraction of oils from wet oleaginous yeast and microalgae under mild conditions. Two ILs, the imidazolium 1-ethyl-3-methylimidazolium ethylsulfate [C2mim][EtSO4] IL and the phosphonium (tetrabutylphosphonium propanoate [P4444][Prop]) IL were assessed for their ability to facilitate extraction of PUFA-containing lipids from a Thraustochytrium sp. (T18) through efficient cell wall disruption. The oil extracted after IL pretreatment was further characterized with respect to fatty acid methyl ester (FAME) composition, while the effects of process parameters, such as the ratio of ionic liquid to co-solvent, the mass ratio of microalgae to the mixture of ionic liquid, and type of co-solvent were also investigated for both ILs. The results indicate that these ILs can disrupt the cells of Thraustochytrium sp. when mixed with a co-solvent (methanol), and facilitated the recovery of oils over a large degree of dewatered Thraustochytrium biomass (0–77.2 wt% water) in a short period of time (60 min) at ambient temperature, hence demonstrating a water compatible, low-energy, lipid recovery method. The lipid recovery was not affected by repeated usage of recycled ILs (tested up to five times).

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Schärer, Lukas, Dagmar Knoflach, Dita B. Vizoso, Gunde Rieger, and Ursula Peintner. "Thraustochytrids as novel parasitic protists of marine free-living flatworms: Thraustochytrium caudivorum sp. nov. parasitizes Macrostomum lignano." Marine Biology 152, no. 5 (July 18, 2007): 1095–104. http://dx.doi.org/10.1007/s00227-007-0755-4.

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26

Takao, Y. "Ecological lilationships among the viruses and thraustochytrids." Journal of Japanese Society for Extremophiles 6, no. 1 (2007): 68–73. http://dx.doi.org/10.3118/jjse.6.68.

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27

Kalidasan, K. "Antioxidant activity of mangrove-derived marine thraustochytrids." Mycosphere 6, no. 5 (2015): 602–11. http://dx.doi.org/10.5943/mycosphere/6/5/9.

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28

Lee Chang, Kim Jye, Maged P. Mansour, Graeme A. Dunstan, Susan I. Blackburn, Anthony Koutoulis, and Peter D. Nichols. "Odd-chain polyunsaturated fatty acids in thraustochytrids." Phytochemistry 72, no. 11-12 (August 2011): 1460–65. http://dx.doi.org/10.1016/j.phytochem.2011.04.001.

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29

TAOKA, Yousuke, Naoki NAGANO, Yuji OKITA, Hitoshi IZUMIDA, Shinichi SUGIMOTO, and Masahiro HAYASHI. "Extracellular Enzymes Produced by Marine Eukaryotes, Thraustochytrids." Bioscience, Biotechnology, and Biochemistry 73, no. 1 (January 23, 2009): 180–82. http://dx.doi.org/10.1271/bbb.80416.

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30

Damare, Varada S. "Diversity of thraustochytrid protists isolated from brown alga, Sargassum cinereum using 18S rDNA sequencing and their morphological response to heavy metals." Journal of the Marine Biological Association of the United Kingdom 95, no. 2 (November 11, 2014): 265–76. http://dx.doi.org/10.1017/s0025315414001696.

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Thraustochytrids, the exclusively marine organisms of kingdom Stramenopila and a source of essential fatty acids in the marine milieu, possess an osmoheterotrophic mode of nutrition and are therefore affected by type and source of available organic matter and pollution. To study their response to heavy metal pollution, they were isolated from the brown alga Sargassum cinereum from the coastal waters of Dona Paula, Goa, India. A total of 22 isolates were obtained from two samples collected during February and March 2012. Based on their 18S rRNA gene sequencing, the majority of the isolates were identified as Thraustochytrium kinnei. The rest were identified to be Sicyoidochytrium minutum, Ulkenia visurgensis and species of Thraustochytrium and Aurantiochytrium. Six isolates were screened for various enzymatic activities. Characteristic and distinctive enzyme profile was obtained from isolates of different genera. All isolates were also screened for their tolerance to heavy metals. They showed good growth in the presence of Mn2+. The other metals that were tolerated by most of the isolates were in the order Ni2+ > Cr6+ > Zn2+. Seven isolates grew in the presence of Cu2+, and six in the presence of Cd2+. The isolates growing on metals showed vast differences from their normal morphology such as small colony size, shrunken cells etc. Scanning electron micrographs revealed holes or depressions in the cell wall in the presence of metals. On the whole, the isolates belonging to Ulkenia visurgensis and Aurantiochytrium sp. showed tolerance to more metals than Thraustochytrium kinnei. Cluster analysis showed no peculiar trend of metal tolerance to any particular genus as the characters were scattered in the clusters.

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31

Fan, K. W., L. L. P. Vrijmoed, and E. B. G. Jones. "Zoospore Chemotaxis of Mangrove Thraustochytrids from Hong Kong." Mycologia 94, no. 4 (July 2002): 569. http://dx.doi.org/10.2307/3761708.

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32

Otagiri, Masato, Ammara Khalid, Shigeharu Moriya, Hiroyuki Osada, and Shunji Takahashi. "Novel squalene-producing thraustochytrids found in mangrove water." Bioscience, Biotechnology, and Biochemistry 81, no. 10 (August 10, 2017): 2034–37. http://dx.doi.org/10.1080/09168451.2017.1359485.

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33

Fan, K. W., L. L. P. Vrijmoed, and E. B. G. Jones. "Zoospore chemotaxis of mangrove thraustochytrids from Hong Kong." Mycologia 94, no. 4 (July 2002): 569–78. http://dx.doi.org/10.1080/15572536.2003.11833185.

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Kanchana, R., Usha Devi Muraleedharan, and Seshagiri Raghukumar. "Alkaline lipase activity from the marine protists, thraustochytrids." World Journal of Microbiology and Biotechnology 27, no. 9 (February 5, 2011): 2125–31. http://dx.doi.org/10.1007/s11274-011-0676-8.

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35

Xie, Yunxuan, Biswarup Sen, and Guangyi Wang. "Mining terpenoids production and biosynthetic pathway in thraustochytrids." Bioresource Technology 244 (November 2017): 1269–80. http://dx.doi.org/10.1016/j.biortech.2017.05.002.

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36

Chamberlain, Anthony H. L., and Stephen T. Moss. "The thraustochytrids: a protist group with mixed affinities." Biosystems 21, no. 3-4 (January 1988): 341–49. http://dx.doi.org/10.1016/0303-2647(88)90031-7.

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37

Du, Fei, Yu-Zhou Wang, Ying-Shuang Xu, Tian-Qiong Shi, Wen-Zheng Liu, Xiao-Man Sun, and He Huang. "Biotechnological production of lipid and terpenoid from thraustochytrids." Biotechnology Advances 48 (May 2021): 107725. http://dx.doi.org/10.1016/j.biotechadv.2021.107725.

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38

Santangelo, Giovanni, Lucia Bongiorni, and Lucrezia Pignataro. "Thraustochytrids (fungoid protist): an unexplored component of marine sediment microbiota." Scientia Marina 68, S1 (April 30, 2004): 43–48. http://dx.doi.org/10.3989/scimar.2004.68s143.

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39

Wang, Qiuzhen, Huike Ye, Yunxuan Xie, Yaodong He, Biswarup Sen, and Guangyi Wang. "Culturable Diversity and Lipid Production Profile of Labyrinthulomycete Protists Isolated from Coastal Mangrove Habitats of China." Marine Drugs 17, no. 5 (May 6, 2019): 268. http://dx.doi.org/10.3390/md17050268.

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Labyrinthulomycete protists have gained significant attention in the recent past for their biotechnological importance. Yet, their lipid profiles are poorly described because only a few large-scale isolation attempts have been made so far. Here, we isolated more than 200 strains from mangrove habitats of China and characterized the molecular phylogeny and lipid accumulation potential of 71 strains. These strains were the closest relatives of six genera namely Aurantiochytrium, Botryochytrium, Parietichytrium, Schizochytrium, Thraustochytrium, and Labyrinthula. Docosahexaenoic acid (DHA) production of the top 15 strains ranged from 0.23 g/L to 1.14 g/L. Two labyrinthulid strains, GXBH-107 and GXBH-215, exhibited unprecedented high DHA production potential with content >10% of biomass. Among all strains, ZJWZ-7, identified as an Aurantiochytrium strain, exhibited the highest DHA production. Further optimization of culture conditions for strain ZJWZ-7 showed improved lipid production (1.66 g/L DHA and 1.68 g/L saturated fatty acids (SFAs)) with glycerol-malic-acid, peptone-yeast-extract, initial pH 7, 28 °C, and rotation rate 150 rpm. Besides, nitrogen source, initial pH, temperature, and rotation rate had significant effects on the cell biomass, DHA, and SFAs production. This study provides the identification and characterization of nearly six dozen thraustochytrids and labyrinthulids with high potential for lipid accumulation.

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Fossier Marchan, Loris, Kim J. Lee Chang, Peter D. Nichols, Wilfrid J. Mitchell, Jane L. Polglase, and Tony Gutierrez. "Taxonomy, ecology and biotechnological applications of thraustochytrids: A review." Biotechnology Advances 36, no. 1 (January 2018): 26–46. http://dx.doi.org/10.1016/j.biotechadv.2017.09.003.

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41

Raghukumar, Chandralata, S. Nagarkar, and S. Raghukumar. "Association of thraustochytrids and fungi with living marine algae." Mycological Research 96, no. 7 (July 1992): 542–46. http://dx.doi.org/10.1016/s0953-7562(09)80978-7.

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42

Abdel-Wahab, Mohamed A., Abd El-Rahim M. A. El-Samawaty, Abdallah M. Elgorban, and Ali H. Bahkali. "Fatty acid production of thraustochytrids from Saudi Arabian mangroves." Saudi Journal of Biological Sciences 28, no. 1 (January 2021): 855–64. http://dx.doi.org/10.1016/j.sjbs.2020.11.024.

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43

Abe, Eriko, Keishi Sakaguchi, Masahiro Hayashi, Daisuke Honda, Kazutaka Ikeda, Ryo Taguchi, Nozomu Okino, and Makoto Ito. "Metabolism and functions of DHA-containing phospholipids in thraustochytrids." Chemistry and Physics of Lipids 164 (August 2011): S34. http://dx.doi.org/10.1016/j.chemphyslip.2011.05.109.

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44

Li, Qian, Xin Wang, Xianhua Liu, Nianzhi Jiao, and Guangyi Wang. "Abundance and Novel Lineages of Thraustochytrids in Hawaiian Waters." Microbial Ecology 66, no. 4 (August 14, 2013): 823–30. http://dx.doi.org/10.1007/s00248-013-0275-3.

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45

Jain, Ruchi, Seshagiri Raghukumar, Kari Sambaiah, Yasuyuki Kumon, and Toro Nakahara. "Docosahexaenoic acid accumulation in thraustochytrids: search for the rationale." Marine Biology 151, no. 5 (January 25, 2007): 1657–64. http://dx.doi.org/10.1007/s00227-007-0608-1.

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46

Bagul, Vaishali P., and Uday S. Annapure. "Isolation of fast-growing thraustochytrids and seasonal variation on the fatty acid composition of thraustochytrids from mangrove regions of Navi Mumbai, India." Journal of Environmental Management 290 (July 2021): 112597. http://dx.doi.org/10.1016/j.jenvman.2021.112597.

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47

Bongiorni, L., and F. Dini. "Distribution and abundance of thraustochytrids in different Mediterranean coastal habitats." Aquatic Microbial Ecology 30 (2002): 49–56. http://dx.doi.org/10.3354/ame030049.

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48

Kimura, Hiroyuki, and Takeshi Naganuma. "Thraustochytrids: A neglected agent of the marine microbial food chain." Aquatic Ecosystem Health & Management 4, no. 1 (April 2001): 13–18. http://dx.doi.org/10.1080/146349801753569243.

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49

Raghukumar, Seshagiri. "Ecology of the marine protists, the Labyrinthulomycetes (Thraustochytrids and Labyrinthulids)." European Journal of Protistology 38, no. 2 (January 2002): 127–45. http://dx.doi.org/10.1078/0932-4739-00832.

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50

Nagano, Naoki, Shou Matsui, Tomoyo Kuramura, Yousuke Taoka, Daiske Honda, and Masahiro Hayashi. "The Distribution of Extracellular Cellulase Activity in Marine Eukaryotes, Thraustochytrids." Marine Biotechnology 13, no. 2 (May 5, 2010): 133–36. http://dx.doi.org/10.1007/s10126-010-9297-8.

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