scholarly journals Algal Lipid Bodies: Stress Induction, Purification, and Biochemical Characterization in Wild-Type and Starchless Chlamydomonas reinhardtii

2009 ◽  
Vol 8 (12) ◽  
pp. 1856-1868 ◽  
Author(s):  
Zi Teng Wang ◽  
Nico Ullrich ◽  
Sunjoo Joo ◽  
Sabine Waffenschmidt ◽  
Ursula Goodenough
Keyword(s):  
Fatty Acids ◽  
Chlamydomonas Reinhardtii ◽  
Unsaturated Fatty Acids ◽  
Biochemical Characterization ◽  
Genetic Manipulation ◽  
Lipid Bodies ◽  
Wild Type ◽  
Soil Alga ◽  
Algal Lipid ◽  
Low Levels

ABSTRACT When the unicellular green soil alga Chlamydomonas reinhardtii is deprived of nitrogen after entering stationary phase in liquid culture, the cells produce abundant cytoplasmic lipid bodies (LBs), as well as abundant starch, via a pathway that accompanies a regulated autophagy program. After 48 h of N starvation in the presence of acetate, the wild-type LB content has increased 15-fold. When starch biosynthesis is blocked in the sta6 mutant, the LB content increases 30-fold, demonstrating that genetic manipulation can enhance LB production. The use of cell wall-less strains permitted development of a rapid “popped-cell” microscopic assay to quantitate the LB content per cell and permitted gentle cell breakage and LB isolation. The highly purified LBs contain 90% triacylglycerol (TAG) and 10% free fatty acids (FFA). The fatty acids associated with the TAGs are ∼50% saturated (C16 and C18) fatty acids and ∼50% unsaturated fatty acids, half of which are in the form of oleic acid (C18:1). The FFA are ∼50% C16 and ∼50% C18. The LB-derived TAG yield from a liter of sta6 cells at 107 cells/ml after starvation for 48 h is calculated to approach 400 mg. The LB fraction also contains low levels of charged glycerolipids, with the same profile as whole-cell charged glycerolipids, that presumably form LB membranes; chloroplast-specific neutral glycerolipids (galactolipids) are absent. Very low levels of protein are also present, but all matrix-assisted laser desorption ionization-identified species are apparent contaminants. Nitrogen stress-induced LB production in C. reinhardtii has the hallmarks of a discrete pathway that should be amenable to additional genetic and culture condition manipulation.

Distinct Modulation of Wild-Type and Selective Gene Mutated Vitamin D Receptor by Essential Poly Unsaturated Fatty Acids

2021 ◽  
Vol 21 ◽  
Author(s):  
Hari Balaji ◽  
Selvaraj Ayyamperuma ◽  
Niladri Saha ◽  
Shyam Sundar Pottabathula ◽  
Jubie Selvaraj ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Vitamin D ◽  
Ligand Binding ◽  
Vitamin D Deficiency ◽  
Binding Affinity ◽  
Unsaturated Fatty Acids ◽  
Binding Domain ◽  
Ligand Binding Domain ◽  
Binding Energies ◽  
Wild Type

: Vitamin-D deficiency is a global concern. Gene mutations in the vitamin D receptor’s (VDR) ligand binding domain (LBD) variously alter the ligand binding affinity, heterodimerization with retinoid X receptor (RXR) and inhibit coactivator interactions. These LBD mutations may result in partial or total hormone unresponsiveness. A plethora of evidence report that selective long chain polyunsaturated fatty acids (PUFAs) including eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and arachidonic acid (AA) bind to the ligand-binding domain of VDR and lead to transcriptional activation. We therefore hypothesize that selective PUFAs would modulate the dynamics and kinetics of VDRs, irrespective bioactive of vitamin-D binding. The spatial arrangements of the selected PUFAs in VDR active site were examined by in-silico docking studies. The docking results revealed that PUFAs have fatty acid structure-specific binding affinity towards VDR. The calculated EPA, DHA & AA binding energies (Cdocker energy) were lesser compared to vitamin-D in wild type of VDR (PDB id: 2ZLC). Of note, the DHA has higher binding interactions to the mutated VDR (PDB id: 3VT7) when compared to the standard Vitamin-D. Molecular dynamic simulation was utilized to confirm the stability of potential compound binding of DHA with mutated VDR complex. These findings suggest the unique roles of PUFAs in VDR activation and may offer alternate strategy to circumvent vitamin-D deficiency.

EXTH-27. MOLECULAR CHARACTERIZATION AND PRECLINICAL DEVELOPMENT OF NOVEL SMALL MOLECULE INHIBITOR SPECIFIC FOR WILD-TYPE IDH1 FOR FERROPTOSIS INDUCTION IN GLIOBLASTOMA

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi169-vi169
Author(s):  
Kevin Murnan ◽  
Serena Tommasini-Ghelfi ◽  
Lisa Hurley ◽  
Corey Dussold ◽  
Daniel Wahl ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Cell Death ◽  
Plasma Membrane ◽  
Fatty Acid ◽  
Unsaturated Fatty Acids ◽  
De Novo ◽  
De Novo Lipogenesis ◽  
Therapeutic Modality ◽  
Wild Type ◽  
Genetic Ablation

Abstract Increased de novo synthesis, mobilization and uptake of fatty acids are required to provide sufficient lipids for membrane biogenesis in support of rapid tumor cell division and growth. In addition to their structural roles as components of the plasma membrane, fatty acid-derived lipids regulate ferroptotic cell death, a type of programmed cell death, when oxidized by iron-dependent lipoxygenase enzymes. De novo lipogenesis and the defense against oxidative lipid damage require large amounts of cytosolic NADPH. Our group has recently found that HGG up-regulate wild-type Isocitrate dehydrogenase 1 (referred to hereafter as ‘wt-IDH1high HGG’) to generate large quantities of cytosolic NADPH. RNAi-mediated knockdown of wt-IDH1, alone and in combination with radiation therapy (RT), slows the growth of patient-derived HGG xenografts, while overexpression of wt-IDH1 promotes intracranial HGG growth. Isotope tracer and liquid chromatography-based lipidomic studies indicated that wt-IDH1 supports the de novo biosynthesis of mono-unsaturated fatty acids (MUFAs) and promotes the incorporation of monounsaturated phospholipids into the plasma membrane, while displacing polyunsaturated fatty acid (PUFA) phospholipids. In addition, enhanced NADPH production in wt-IDH1high HGG increases glutathione (GSH) level, reduces reactive oxygen species (ROS), activates the phospholipid peroxidase glutathione peroxidase 4 (GPX4)-driven lipid repair pathway, and dampens the accumulation of PUFA-containing lipid peroxides, known executioners of ferroptosis. To pharmacologically target wt-IDH1,we have used and characterized wt-IDH1i-13, a first-in-class competitive α,β-unsaturated enone (AbbVie). wt-IDH1i-13 potently inhibits wt-IDH1 enzymatic activity, by covalently binding to the NADP+ binding pocket. Our data indicate that wt-IDH1i-13 promotes ferroptosis, which can be rescued by pre-treatment of cells with the peroxyl scavenger and ferroptosis inhibitor ferrostatin. wt-IDH1i-13 is brain-penetrant, and similar to genetic ablation, reduces progression and extends the survival of wt-IDH1high HGG bearing mice, alone and in combination with RT. These studies credential to wt-IDH1i-13 as a novel therapeutic modality for the treatment of wt-IDH1 gliomas.

scholarly journals Chlamydomonas reinhardtii Is a Potential Food Supplement with the Capacity to Outperform Chlorella and Spirulina

2020 ◽  
Vol 10 (19) ◽  
pp. 6736
Author(s):  
Randa Darwish ◽  
Mohamed A. Gedi ◽  
Patchaniya Akepach ◽  
Hirut Assaye ◽  
Abdelrahman S. Zaky ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Chlamydomonas Reinhardtii ◽  
Unsaturated Fatty Acids ◽  
Model Organism ◽  
Dry Weight ◽  
Essential Amino Acids ◽  
Food Supplement ◽  
Alpha Linolenic Acid ◽  
Total Carotenoids ◽  
Total Fatty Acids

Chlamydomonas reinhardtii is a green microalgae used as a model organism associated with biotechnological applications, yet its nutritional value has not been assessed. This study investigates the nutritional capacity of C. reinhardtii as an additional value for this species beyond its known potential in biofuels and bio-products production. The composition of key nutrients in C. reinhardtii was compared with Chlorella and Spirulina, the species widely regarded as a superfood. The results revealed that the protein content of C. reinhardtii (46.9%) was comparable with that of Chlorella (45.3) and Spirulina (50.4%) on a dry weight basis. C. reinhardtii contained all the essential amino acids with good scores based on FAO/WHO values (0.9–1.9) as in Chlorella and Spirulina. Unsaturated fatty acids predominated the total fatty acids profile of C. reinhardtii were ~74 of which ~48% are n-3 fatty acids. Alpha-linolenic acid (ALA) content in C. reinhardtii (42.4%) was significantly higher than that of Chlorella (23.4) and Spirulina (0.12%). For minerals, Spirulina was rich in iron (3.73 mg/g DW) followed by Chlorella (1.34 mg/g DW) and C. reinhardtii (0.96 mg/g DW). C. reinhardtii, unlike the other two species, consisted of selenium (10 µg/g DW), and had a remarkably lower heavy metal load. Moreover, C. reinhardtii contained relatively high concentrations of chlorophyll (a + b) and total carotenoids (28.6 mg/g DW and 6.9 mg/g DW, respectively) compared with Chlorella (12.0 mg/g DW and 1.8 mg/g DW, respectively) and Spirulina (8.6 mg/g DW and 0.8 mg/g DW, respectively). This study confirms that, based on its nutrient credentials, C. reinhardtii has great potential as a new superfood or ingredient for a food supplement.

scholarly journals Biochemical Characterization of Wild-Type and Mutant Isoamylases of Chlamydomonas reinhardtii Supports a Function of the Multimeric Enzyme Organization in Amylopectin Maturation

2001 ◽  
Vol 125 (4) ◽  
pp. 1723-1731 ◽  
Author(s):  
David Dauvillée ◽  
Christophe Colleoni ◽  
Gregory Mouille ◽  
Matthew K. Morell ◽  
Christophe d'Hulst ◽  
...  
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Biochemical Characterization ◽  
Wild Type ◽  
Enzyme Organization

scholarly journals Two Lipid Signals Guide Fruiting Body Development of Myxococcus xanthus

mBio ◽  
2014 ◽  
Vol 5 (1) ◽  
Author(s):  
Swapna Bhat ◽  
Tilman Ahrendt ◽  
Christina Dauth ◽  
Helge B. Bode ◽  
Lawrence J. Shimkets
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Fruiting Body ◽  
Myxococcus Xanthus ◽  
Chain Fatty Acid ◽  
Fatty Alcohols ◽  
Lipid Bodies ◽  
Wild Type ◽  
Bioactive Lipids ◽  
Content Type

ABSTRACTMyxococcus xanthusproduces several extracellular signals that guide fruiting body morphogenesis and spore differentiation. Mutants defective in producing a signal may be rescued by codevelopment with wild-type cells or cell fractions containing the signal. In this paper, we identify two molecules that rescue development of the E signal-deficient mutant LS1191 at physiological concentrations,iso15:0 branched-chain fatty acid (FA) and 1-iso15:0-alkyl-2,3-di-iso15:0-acyl glycerol (TG1), a development-specific monoalkyl-diacylglycerol. The physiological concentrations of the bioactive lipids were determined by mass spectrometry from developing wild-type cells using chemically synthesized standards. Synthetic TG1 restored fruiting body morphogenesis and sporulation and activated the expression of the developmentally regulated gene with locus tagMXAN_2146at physiological concentrations, unlike its nearly identical tri-iso15:0 triacylglycerol (TAG) counterpart, which has an ester linkage instead of an ether linkage.iso15:0 FA restored development at physiological concentrations, unlike palmitic acid, a straight-chain fatty acid. The addition of either lipid stimulates cell shortening, with an 87% decline in membrane surface area, concomitantly with the production of lipid bodies at each cell pole and in the center of the cell. We suggest that cells produce triacylglycerol from membrane phospholipids. Bioactive lipids may be released byprogrammedcelldeath (PCD), which claims up to 80% of developing cells, since cells undergoing PCD produce lipid bodies before lysing.IMPORTANCELike mammalian adipose tissue, many of theM. xanthuslipid body lipids are triacylglycerols (TAGs), containing ester-linked fatty acids. In both systems, ester-linked fatty acids are retrieved from TAGs with lipases and consumed by the fatty acid degradation cycle. Both mammals andM. xanthusalso produce lipids containing ether-linked fatty alcohols with alkyl or vinyl linkages, such as plasmalogens. Alkyl and vinyl linkages are not hydrolyzed by lipases, and no clear role has emerged for lipids bearing them. For example, plasmalogen deficiency in mice has detrimental consequences to spermatocyte development, myelination, axonal survival, eye development, and long-term survival, though the precise reasons remain elusive. Lipids containing alkyl- and vinyl-linked fatty alcohols are development-specific products inM. xanthus. Here, we show that one of them rescues the development of E signal-producing mutants at physiological concentrations.

scholarly journals Biochemical Characterization by GC-MS of Palm Kernel Oils Produced in Côte d’Ivoire

2021 ◽  
Vol 10 (3) ◽  
pp. 33
Author(s):  
Ahou Irène Kouadio
Keyword(s):  
Fatty Acids ◽  
Biochemical Composition ◽  
Unsaturated Fatty Acids ◽  
Biochemical Characterization ◽  
Elaeis Guineensis ◽  
Saturated Fatty Acids ◽  
Palm Kernel ◽  
Alpha Tocopherol ◽  
Undecylenic Acid ◽  
Human Diet

The objective of this study was to determine biochemical composition of palm kernel oils produced and consuming in Côte d’Ivoire in order to find out those more suitable for human diet. Our preliminary investigations showed that palm kernel oils consumed in Côte d’Ivoire were those extracted from varieties Dura and Tenera of oil palm (Elaeis guineensis Jacq.) Thus, the types of oils analyzed in this study were oil extracted from the variety Dura (OD) and oil extrated from the variety Tenera (OT). The GC-MS was used to determine the biochemical composition of these oils. The results obtained show that in each oil, seventeen fatty acids were identified by GC–MS. However, among these fatty acids, undecylenic acid was identified only in OD and heptanoic acid was identified only in OT. The two types of oil are rich in saturated fatty acids. However, OD had a relatively higher unsaturated fatty acids content. For the other compounds identified, OT had significantly the highest contents of polyphenols, α-tocopherol and sterols with the predominance of β-sitosterol. These results support that palm kernel oil extracted from the variety Tenera is rich in natural compounds that could be developed as nutraceuticals and phytomedicine. However, some unexpected compounds such as lactones were also identified in the two types of oils. Moreover, it is noted that these lactones were more abundant in oil extracted from the variety Dura (OD).

Sensitivity of Saccharomyces cerevisiae to tannic acid is due to iron deprivation

2001 ◽  
Vol 47 (4) ◽  
pp. 290-293 ◽  
Author(s):  
T Wauters ◽  
D Iserentant ◽  
H Verachtert
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Fatty Acids ◽  
Growth Rate ◽  
Tannic Acid ◽  
Unsaturated Fatty Acids ◽  
Acid Resistance ◽  
Maximal Growth ◽  
Wild Type ◽  
Iron Deprivation ◽  
Maximal Growth Rate

Tannic acid inhibited the growth of the yeast Saccharomyces cerevisiae. Growth medium supplementation with more nitrogen or metal ions showed that only iron ions could restore the maximal growth rate of S. cerevisiae. Tannic acid resistant mutants were previously isolated by screening for tannic acid resistance and were all cytoplasmic petite mutants. While the wild type was very sensitive to iron deprivation conditions when grown in aerobic conditions, the mutants, whether grown aerobically or anaerobically, showed the same growth rate under iron-limited conditions as under iron-repleted conditions. Also, the wild type grown anaerobically was not affected by iron-limited conditions. Cytoplasmic petite mutants obtained by ethidium bromide mutagenesis behaved like the other mutants. During iron limitation, the wild type showed a reduced oxygen uptake rate. Maximal growth rate of the wild type in iron-limited conditions could be restored by the addition to the media of unsaturated fatty acids and sterol. Iron deprivation caused by tannic acid may thus affect the synthesis of a functional respiratory chain as well as the synthesis of unsaturated fatty acids and (or) sterol.Key words: Saccharomyces cerevisiae, tannic acid resistance, iron deprivation, cytoplasmic petite mutant.

scholarly journals Cytoplasmic lipid bodies of neutrophils: formation induced by cis-unsaturated fatty acids and mediated by protein kinase C.

1991 ◽  
Vol 113 (1) ◽  
pp. 137-146 ◽  
Author(s):  
P F Weller ◽  
S W Ryeom ◽  
S T Picard ◽  
S J Ackerman ◽  
A M Dvorak
Keyword(s):  
Fatty Acids ◽  
Protein Kinase C ◽  
Fatty Acid ◽  
Protein Kinase ◽  
Unsaturated Fatty Acids ◽  
Lipid Body ◽  
Lipid Bodies ◽  
Myristate Acetate ◽  
Body Formation ◽  
Kinase C

Lipid bodies, nonmembrane-bound cytoplasmic inclusions, serve as repositories of esterified arachidonate and are increased in cells associated with inflammatory reactions. We have evaluated stimuli and mechanisms responsible for lipid body formation within human polymorphonuclear leukocytes (PMNs). Arachidonic acid and oleic acid stimulated dose-dependent formation of lipid bodies over 0.5-1 h. Other C20 and C18 fatty acids were less active and demonstrated rank orders as follows: cis-unsaturated fatty acids were much more active than trans-fatty acids, and activity diminished with decreasing numbers of double bonds. Lipid bodies elicited in vitro with cis-fatty acids were ultrastructurally identical to lipid bodies present in PMNs in vivo. Lipid body induction was not because of fatty acid-elicited oxidants or fatty acid-induced ATP depletion. Cis-fatty acid-induced activation of protein kinase C (PKC) was involved in lipid body formation as evidenced by the capacity of other PKC activators, 1-oleoyl-2-acetyl-glycerol and two active phorbol esters, phorbol myristate acetate, and phorbol 12,13 dibutyrate, but not an inactive phorbol, to induce lipid body formation. The PKC inhibitor, 1-O-hexadecyl-2-O-methyl-glycerol, inhibited PMN lipid body formation induced by oleic and arachidonic acids and by 1-oleoyl-2-acetyl-glycerol and phorbol myristate acetate. Other PKC inhibitors (staurosporine, H-7) also inhibited lipid body formation. Formation of lipid bodies in PMNs is a specific cellular response, stimulated by cis-fatty acids and diglycerides and apparently mediated by PKC, which results in the mobilization and deposition of lipids within discrete, ultrastructurally defined cytoplasmic domains.

scholarly journals BIOCHEMICAL CHARACTERIZATION OF Ferula rutbaensis MEDICINAL PLANT IN IRAQI WESTERN DESERT

2020 ◽  
Vol 17 (36) ◽  
pp. 831-844
Author(s):  
Kutayba Farhan DAWOOD ◽  
Ayoob Obaid ALFALAHI ◽  
Shamil Ismail NEAMAH ◽  
Omar Mahmood DHANNOON
Keyword(s):  
Fatty Acids ◽  
Bioactive Compounds ◽  
Unsaturated Fatty Acids ◽  
Biochemical Characterization ◽  
Western Desert ◽  
Morphological Identification ◽  
Plant Parts ◽  
Methanolic Extracts ◽  
Wide Range

Plants used in folk medicine not only represent rich sources for therapeutic materials, but it also plays a crucial role in developing completely or partially novel synthesized drugs. Mharut plant (Ferula rutbaensis) is an integral part of Bedouin therapeutic practices in the western desert of Anbar province-Iraq. Still, to date, this is the first study describing its phytochemical constituents. The plant was growing near the Iraq-Saudi Arabia borders and adapted to a wide range of soils. Traditionally, F. rutbaensis has been widely used to treat acne, stomach and bowel disorders, food poisoning and respiratory problems. Fresh plant samples were collected and morphologically characterized. Likewise, the ITS-based DNA barcoding technique was efficiently used to approve the morphological identification of F. rutbaensis. The GC-MS spectrum was adopted in the phytochemical characterization of aqueous and methanol extracts of fresh and dry plant parts. The aqueous extract of dry roots was the richest source for bioactive compounds than fresh or methanolic extracts of either fresh or dry plant parts. In general, the detected phytochemicals falling into fatty acids, terpenes, hydrocarbon alkanes, and esters. Notably, fatty acids in Oleic and Palmitic acids were the two most abundant bioactive compounds in both aqueous and methanolic extracts of plant fresh and dry roots. The detected unsaturated fatty acids and/or other bioactive components are laying behind the therapeutic properties of F. rutbaensis that can be useful ingredients to prepare Mharut-based cosmetics such as medical soaps, body lotions, skin conditioners and sunscreens. Additionally, some other components were found to have anti-inflammatory, antioxidants, and antimicrobial properties. Further investigations will be necessary to confirm the antimicrobial activity of F. rutbaensis extracts.

scholarly journals Amino Acid Substitutions in the Pore Helix of GluR6 Control Inhibition by Membrane Fatty Acids

2008 ◽  
Vol 132 (1) ◽  
pp. 85-99 ◽  
Author(s):  
Timothy J. Wilding ◽  
Elisabeth Fulling ◽  
Yun Zhou ◽  
James E. Huettner
Keyword(s):  
Fatty Acids ◽  
Unsaturated Fatty Acids ◽  
Kainate Receptors ◽  
Kainate Receptor ◽  
Strong Inhibition ◽  
Wild Type ◽  
Hek 293 ◽  
Channel Inhibition ◽  
293 Cells ◽  
Fatty Acid Inhibition

RNA editing at the Q/R site in the GluR5 and GluR6 subunits of neuronal kainate receptors regulates channel inhibition by lipid-derived modulators including the cis-unsaturated fatty acids arachidonic acid and docosahexaenoic acid. Kainate receptor channels in which all of the subunits are in the edited (R) form exhibit strong inhibition by these compounds, whereas wild-type receptors that include a glutamine (Q) at the Q/R site in one or more subunits are resistant to inhibition. In the present study, we have performed an arginine scan of residues in the pore loop of the GluR6(Q) subunit. Amino acids within the range from −19 to +7 of the Q/R site of GluR6(Q) were individually mutated to arginine and the mutant cDNAs were expressed as homomeric channels in HEK 293 cells. All but one of the single arginine substitution mutants yielded functional channels. Only weak inhibition, typical of wild-type GluR6(Q) channels, was observed for substitutions +1 to +6 downstream of the Q/R site. However, arginine substitution at several locations upstream of the Q/R site resulted in homomeric channels exhibiting strong inhibition by fatty acids, which is characteristic of homomeric GluR6(R) channels. Based on homology with the pore loop of potassium channels, locations at which R substitution induces susceptibility to fatty acid inhibition face away from the cytoplasm toward the M1 and M3 helices and surrounding lipids.

Sign in / Sign up

Export Citation Format

Share Document