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Characterization of an Oleaginous Unicellular Green Microalga, Lobosphaera incisa (Reisigl, 1964) Strain K-1, Isolated From a Tidal Flat in the Yellow Sea, Republic of Korea.

ABSTRACT: Microalgae are considered as sustainable resources for biofuel production. However, recently the focus on microalgal research has shifted toward the investigation of high-value metabolites for potential pharmaceutical and nutritional applications. Herein, we report the identification of a novel oleaginous green microalga isolated from the Yellow Sea in Korea. We also describe the morphological, molecular, and biochemical characteristics of this microalga. On the basis of microscopic and genetic analyses, the isolate was classified as Lobosphaera incisa (the strain was designated as K-1), and molecular phylogeny revealed that the isolate distinctly differed from the other known L. incisa strains. The microalga could be cultivated in various commercial culture media under a relatively broad range of pH and temperature conditions. We also did a rough and detailed estimation of the different cellular components in the microalga. The composition of arachidonic acid (C20:4ω6) in the lipids of L. incisa strain K-1 was relatively high, similar to that in other strains, however, the K-1 strain had higher proportions of the ω3 series of fatty acids (FAs), including α-linolenic acid (C18:3ω3) and eicosapentaenoic acid (C20:5ω3), highlighting its uniqueness and strong potential for biotechnological application. To the best of our knowledge, this is the first report on the isolation of L. incisa from Korea as well as from a marine environment; this novel strain might be useful for the production of high-value ω3 and ω6 polyunsaturated fatty acids (PUFAs).

SUBMITTER: Lee S

PROVIDER: S-EPMC6139321 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundLobosphaera incisa, formerly known as Myrmecia incisa and then Parietochloris incisa, is an oleaginous unicellular green alga belonging to the class Trebouxiophyceae (Chlorophyta). It is the richest known plant source of arachidonic acid, an ω-6 poly-unsaturated fatty acid valued by the pharmaceutical and baby-food industries. It is therefore an organism of high biotechnological interest, and we recently reported the sequence of its chloroplast genome.ResultsWe now report the complete sequence of the mitochondrial genome of L. incisa from high-throughput Illumina short-read sequencing. The circular chromosome of 69,997 bp is predicted to encode a total of 64 genes, some harboring specific self-splicing group I and group II introns. Overall, the gene content is highly similar to that of the mitochondrial genomes of other Trebouxiophyceae, with 34 protein-coding, 3 rRNA, and 27 tRNA genes. Genes are distributed in two clusters located on different DNA strands, a bipartite arrangement that suggests expression from two divergent promoters yielding polycistronic primary transcripts. The L. incisa mitochondrial genome contains families of intergenic dispersed DNA repeat sequences that are not shared with other known mitochondrial genomes of Trebouxiophyceae. The most peculiar feature of the genome is a repetitive palindromic repeat, the LIMP (L. Incisa Mitochondrial Palindrome), found 19 times in the genome. It is formed by repetitions of an AACCA pentanucleotide, followed by an invariant 7-nt loop and a complementary repeat of the TGGTT motif. Analysis of the genome sequencing reads indicates that the LIMP can be a substrate for large-scale genomic rearrangements. We speculate that LIMPs can act as origins of replication. Deep sequencing of the L. incisa transcriptome also suggests that the LIMPs with long stems are sites of transcript processing. The genome also contains five copies of a related palindromic repeat, the HyLIMP, with a 10-nt motif related to that of the LIMP.ConclusionsThe mitochondrial genome of L. incisa encodes a unique type of repetitive palindromic repeat sequence, the LIMP, which can mediate genome rearrangements and play a role in mitochondrial gene expression. Experimental studies are needed to confirm and further characterize the functional role(s) of the LIMP.

Project description:The green microalga Lobosphaera incisa accumulates triacylglycerols (TAGs) with exceptionally high levels of long-chain polyunsaturated fatty acid (LC-PUFA) arachidonic acid (ARA) under nitrogen (N) deprivation. Phosphorous (P) deprivation induces milder changes in fatty acid composition, cell ultrastructure, and growth performance. We hypothesized that the resource-demanding biosynthesis and sequestration of ARA-rich TAG in lipid droplets (LDs) are associated with the enhancement of catabolic processes, including membrane lipid turnover and autophagic activity. Although this work focuses mainly on N deprivation, a comparative analysis of N and P deprivation responses is included. The results of lipidomic profiling showed a differential impact of N and P deprivation on the reorganization of glycerolipids. The formation of TAG under N deprivation was associated with the enhanced breakdown of chloroplast glycerolipids and the formation of lyso-lipids. N-deprived cells displayed a profound reorganization of cell ultrastructure, including internalization of cellular material into autophagic vacuoles, concomitant with the formation of LDs, while P-deprived cells showed better cellular ultrastructural integrity. The expression of the hallmark autophagy protein ATG8 and the major lipid droplet protein (MLDP) genes were coordinately upregulated, but to different extents under either N or P deprivation. The expression of the Δ5-desaturase gene, involved in the final step of ARA biosynthesis, was coordinated with ATG8 and MLDP, exclusively under N deprivation. Concanamycin A, the inhibitor of vacuolar proteolysis and autophagic flux, suppressed growth and enhanced levels of ATG8 and TAG in N-replete cells. The proportions of ARA in TAG decreased with a concomitant increase in oleic acid under both N-replete and N-deprived conditions. The photosynthetic apparatus's recovery from N deprivation was impaired in the presence of the inhibitor, along with the delayed LD degradation. The GFP-ATG8 processing assay showed the release of free GFP in N-replete and N-deprived cells, supporting the existence of autophagic flux. This study provides the first insight into the homeostatic role of autophagy in L. incisa and points to a possible metabolic link between autophagy and ARA-rich TAG biosynthesis.

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Characterization of an Oleaginous Unicellular Green Microalga, Lobosphaera incisa (Reisigl, 1964) Strain K-1, Isolated From a Tidal Flat in the Yellow Sea, Republic of Korea.

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