Highly efficient transgene-free targeted mutagenesis and single-stranded oligodeoxynucleotide-mediated precise knock-in in the industrial microalga Euglena gracilis using Cas9 ribonucleoproteins.

Authors: Toshihisa Nomura, Komaki Inoue, Yukiko Uehara-Yamaguchi...

Topics: CRISPR-Cas Systems; Euglena gracilis; Gene Editing; Gene Knock-In Techniques; Microalgae; Mutagenesis; Oligodeoxyribonucleotides; Ribonucleoproteins

PubMed: 31131534
DOI: 10.1111/pbi.13174

Authors: Laura Barsanti, Paolo Gualtieri

Algae (macro- and micro-algae) can be defined as light-driven cell factories that synthesize bioactive compounds consisting of primary metabolites (i [...].

Topics: Euglena gracilis; Glucans

PubMed: 36982916
DOI: 10.3390/ijms24065844

Authors: Magnus Alm Rosenblad, Marcela Dávila López, Tore Samuelsson...

The ribonucleoprotein RNase MRP is responsible for the processing of ribosomal RNA precursors. It is found in virtually all eukaryotes that have been examined. In the Euglenozoa, including the genera Euglena, Diplonema and kinetoplastids, MRP RNA and protein subunits have so far escaped detection using bioinformatic methods. However, we now demonstrate that the RNA component is widespread among the Euglenozoa and that these RNAs have secondary structures that conform to the structure of all other phylogenetic groups. In Euglena, we identified the same set of P/MRP protein subunits as in many other protists. However, we failed to identify any of these proteins in the kinetoplastids. This finding poses interesting questions regarding the structure and function of RNase MRP in these species.

Topics: Base Pairing; Base Sequence; DNA, Kinetoplast; Endoribonucleases; Euglena; Kinetoplastida; Nucleic Acid Conformation; Phylogeny; Protozoan Proteins; RNA Processing, Post-Transcriptional; RNA, Protozoan

PubMed: 34308760
DOI: 10.1080/15476286.2021.1952758

Authors: Martin Zoltner, Mark C Field

is a unicellular photosynthetic eukaryotic flagellate of the Discoba supergroup, which also encompasses Kinetoplastida and Diplonema. Plastids have green algal origin and are secondarily acquired. The nuclear genome is extremely large and many genes suggest multiple endosymbiotic/gene transfer events, i.e. derivation from prokaryotes of various lineages. is remarkably robust and can proliferate in environments contaminated with heavy metals and acids. Extraordinary metabolic plasticity and a mixotrophic lifestyle confers an ability to thrive in a broad range of environments, as well as facilitating production of many novel metabolites, making of considerable biotechnological importance.

Topics: Chlorophyta; Euglena gracilis; Photosynthesis; Plastids; Symbiosis

PubMed: 36178464
DOI: 10.1099/mic.0.001241

Authors: Pingwei Gao, Yujie Zhong, Chengfu Sun...

Diverse splicing types in nuclear and chloroplast genes of protist have been recognized for decades. However, the splicing machinery responsible for processing nuclear precursor messenger RNA introns, including -splicing of the 5' terminal outron and spliced leader (SL) RNA, remains elusive. Here, we identify 166 spliceosomal protein genes and two snRNA genes from . by performing bioinformatics analysis from a combination of next-generation and full-length transcriptomic RNA sequencing (RNAseq) data as well as draft genomic data. With the spliceosomal proteins we identified in hand, the insensitivity of . to some splicing modulators is revealed at the sequence level. The prevalence of SL RNA-mediated -splicing is estimated to be more than 70% from our full-length RNAseq data. Finally, the splicing proteomes between . and its three evolutionary cousins within the same Excavata group are compared. In conclusion, our study characterizes the spliceosomal components in . and provides the molecular basis for further exploration of underlying splicing mechanisms.

Topics: Euglena gracilis; Transcriptome; Base Sequence; Spliceosomes; Genomics

PubMed: 37705346
DOI: 10.3724/abbs.2023143

Authors: Mahfuzur R Shah, Erin N Morrison, Adam J Noble...

is a source of high-value natural products. A major factor affecting consistent production of biomass is strain stability. Cryopreservation is a leading strategy for cell-line storage that helps ensure process reproducibility. We developed a simple cryopreservation protocol for heterotrophically cultured that enables the recovery of cells after 1 year with a cell viability of ≅80%. This protocol is suitable for labs interested in the long-term preservation of heterotrophic cultures of and related species.

Topics: Biomass; Cryopreservation; Euglena; Euglena gracilis; Reproducibility of Results

PubMed: 34977684
DOI: 10.1016/j.xpro.2021.101043

Authors: Anna M G Novák Vanclová, Martin Zoltner, Steven Kelly...

Euglena spp. are phototrophic flagellates with considerable ecological presence and impact. Euglena gracilis harbours secondary green plastids, but an incompletely characterised proteome precludes accurate understanding of both plastid function and evolutionary history. Using subcellular fractionation, an improved sequence database and MS we determined the composition, evolutionary relationships and hence predicted functions of the E. gracilis plastid proteome. We confidently identified 1345 distinct plastid protein groups and found that at least 100 proteins represent horizontal acquisitions from organisms other than green algae or prokaryotes. Metabolic reconstruction confirmed previously studied/predicted enzymes/pathways and provided evidence for multiple unusual features, including uncoupling of carotenoid and phytol metabolism, a limited role in amino acid metabolism, and dual sets of the SUF pathway for FeS cluster assembly, one of which was acquired by lateral gene transfer from Chlamydiae. Plastid paralogues of trafficking-associated proteins potentially mediating fusion of transport vesicles with the outermost plastid membrane were identified, together with derlin-related proteins, potential translocases across the middle membrane, and an extremely simplified TIC complex. The Euglena plastid, as the product of many genomes, combines novel and conserved features of metabolism and transport.

Topics: Biological Evolution; Euglena gracilis; Gene Expression Regulation; Gene Transfer, Horizontal; Plastids; Proteins; Proteome

PubMed: 31580486
DOI: 10.1111/nph.16237

Authors: Tengteng Ji, Bing Fang, Yutong Jin...

Obesity, a global disease, can lead to different chronic diseases and a series of social health problems. Lifestyle changes, especially dietary changes, are the most effective way to treat obesity. , a novel food, has attracted much attention. Previous studies have shown that is an important modulator of the host immune response. In this study, the effects of as a nutritional intervention in high-fat-diet-induced obese C57BL/6J mice were investigated regarding adipose tissue accumulation and lipid and glucose metabolism by gavage at the dose of 100 mg/kg bodyweight for 9 weeks. This study is one of the few to investigate, in detail, the preventive effects of dietary on obesity. Five-week-old male C57BL/6J mice were fed with a high-fat diet (HFD) to induce obesity. An obesity model was created by feeding the high-fat diet for a period of 10 weeks. Obese mice were randomized into 2 groups with the same mean body weight, and no significant differences were observed between the groups: (1) the mice in the HEG group were maintained on a high-fat diet and daily gavaged with (100 mg/kg body weight) dissolved in saline ( = 7); and (2) the mice in the HFD group were maintained on a high-fat diet and daily gavaged with saline with the same volume ( = 7). The experiment finished after a nine-week period. The results showed that could reduce the accumulation of white body fat, including subcutaneous fat and visceral fat, and mainly targeted subcutaneous fat. also reduced adipocyte particle size expansion, promoted lipolysis in adipose (adipose triglyceride lipase and hormone-sensitive triglyceride lipase) and liver tissue (reduced non-esterified fatty acid content), and improved obesity-induced ectopic fat deposition and glucose tolerance. Our findings suggest that , as a nutritional intervention in HFDs, efficiently reduces body weight and white adipose tissue deposition. The mechanism of is mainly though enhancing lipolysis. It is worth noting that β-glucan recovers the hyperglycemia and accumulation of ectopic fat within the liver induced by HFD. Our study is one of the few studies to report in detail the preventive effects of dietary on obesity in vivo. This study revealed that also has an important ameliorative effect on obesity and metabolic disorders, which laid a theoretical foundation for its future application in functional foods.

Topics: Animals; Obesity; Diet, High-Fat; Male; Mice, Inbred C57BL; Glucose Intolerance; Mice; Euglena gracilis; Blood Glucose; Disease Models, Animal; Lipid Metabolism; Adipose Tissue

PubMed: 39519613
DOI: 10.3390/nu16213780

Authors: Qing Shao, Lang Hu, Huan Qin...

Euglena, a new superfood on the market, is a nutrient-rich, green single-celled microorganism that features the characteristics of both plant and animal. When cultivated under different conditions, Euglena produces different bioactive nutrients. Interestingly, Euglena is the only known microorganism whose chloroplasts are easy to lose under stress and become permanently bleached. We applied gas chromatography-mass spectrometry (GC-MS) to determine the metabolomes of wild-type (WT) Euglena gracilis and its bleached mutant OflB2 under light stimulation. We found a significant metabolic difference between WT and OflB2 cells in response to light. An increase of membrane components (phospholipids and acylamides) was observed in WT, while a decrease of some stimulant metabolites was detected in OflB2. These metabolomic changes after light stimulation are of great significance to the development of Euglena chloroplasts and their communications with the nucleus.

Topics: Cluster Analysis; Euglena gracilis; Light; Metabolome; Metabolomics; Mutation; Pigments, Biological; Principal Component Analysis

PubMed: 31697795
DOI: 10.1371/journal.pone.0224926

Authors: Masami Nakazawa, Hiroko Ando, Ayusa Nishimoto...

In Euglena gracilis, wax ester fermentation produces ATP during anaerobiosis. Here, we report that anaerobic wax ester production is suppressed when the mitochondrial electron transport chain complex I is inhibited by rotenone, whereas it is increased by the uncoupler carbonyl cyanide m-chlorophenylhydrazone (CCCP). The ADP/ATP ratio in anaerobic cells is elevated by treatment with either rotenone or CCCP. Gene silencing experiments indicate that acyl-CoA dehydrogenase, electron transfer flavoprotein (ETF), and rhodoquinone (RQ) participate in wax ester production. These results suggest that fatty acids are synthesized in mitochondria by the reversal of β-oxidation, where trans-2-enoyl-CoA is reduced mainly by acyl-CoA dehydrogenase using the electrons provided by NADH via the electron transport chain complex I, RQ, and ETF, and that ATP production is highly supported by anaerobic respiration utilizing trans-2-enoyl-CoA as a terminal electron acceptor.

Topics: Acyl-CoA Dehydrogenase; Adenosine Diphosphate; Adenosine Triphosphate; Anaerobiosis; Cell Respiration; Esters; Euglena gracilis; Fatty Acids; Fermentation; Mitochondria; RNA Interference; Rotenone; Uncoupling Agents; Waxes

PubMed: 30328102
DOI: 10.1002/1873-3468.13276