Lipid Biosynthesis Pathways Research Articles

Transcriptional Regulation of T-Cell Lipid Metabolism: Implications for Plasma Membrane Lipid Rafts and T-Cell Function.

It is well established that cholesterol and glycosphingolipids are enriched in the plasma membrane (PM) and form signaling platforms called lipid rafts, essential for T-cell activation and function. Moreover, changes in PM lipid composition affect the biophysical properties of lipid rafts and have a role in defining functional T-cell phenotypes. Here, we review the role of transcriptional regulators of lipid metabolism including liver X receptors α/β, peroxisome proliferator-activated receptor γ, estrogen receptors α/β (ERα/β), and sterol regulatory element-binding proteins in T-cells. These receptors lie at the interface between lipid metabolism and immune cell function and are endogenously activated by lipids and/or hormones. Importantly, they regulate cellular cholesterol, fatty acid, glycosphingolipid, and phospholipid levels but are also known to modulate a broad spectrum of immune responses. The current evidence supporting a role for lipid metabolism pathways in controlling immune cell activation by influencing PM lipid raft composition in health and disease, and the potential for targeting lipid biosynthesis pathways to control unwanted T-cell activation in autoimmunity is reviewed.

Vegetable oil production in vegetative plant tissues

Vegetable oils, which are stored in seeds as triacylglycerol (TAG), represent a valuable source of food, fuel, and industrial raw materials. Given the increasing demand for vegetable oils, it is essential to increase vegetable oil production in a sustainable manner. To boost the supply of vegetable oil, strategies to produce oils in the vegetative tissues of biomass crops are being investigated. Producing oils in leaves is challenging, due to the major role of leaves in producing sugars, via photosynthesis, that are then transported to storage tissues. Several strategies have been developed to overcome this limitation, such as increasing fatty acid and TAG biosynthesis, stabilizing lipid droplets, and reducing carbon flux to starch biosynthesis in the leaves. For example, overexpressing the WRINKLED 1 (WRI1) transcription factor, which upregulates fatty acid biosynthesis; DIACYLGLYCEROL ACYLTRANSFERASE 1 (DGAT1) enzymes, which participate in the last step of TAG biosynthesis; and oleosin, which stabilizes oil droplets and simultaneously suppressing the expression of genes encoding enzymes involved in lipogenesis and lipases led to the accumulation of significant levels of oil in leaves in tobacco, Arabidopsis, and sugarcane. To facilitate the sustainable production of biofuels and industrial raw materials, efforts are underway to modulate metabolic flux between the carbohydrate and lipid biosynthesis pathways in non-edible biomass plants that can grow on barren or reclaimed lands.

Mutants of Yarrowia lipolytica NCIM 3589 grown on waste cooking oil as a biofactory for biodiesel production

BackgroundOleaginous yeasts are fast emerging as a possible feedstock for biodiesel production. Yarrowia lipolytica, a model oleaginous yeast is known to utilize a variety of hydrophobic substrates for lipid accumulation including waste cooking oil (WCO). Approaches to increase lipid content in this yeast include metabolic engineering which requires manipulation of multiple genes in the lipid biosynthesis pathway. A classical and cost-effective approach, namely, random chemical mutagenesis on the yeast can lead to increased production of biodiesel as is explored here.ResultsIn this study, chemical mutagenesis using the alkylating agent, N- methyl-N′-nitro-N-nitrosoguanidine (MNNG) as well as an additional treatment with cerulenin, a fatty acid synthase inhibitor generated 800 mutants of Y. lipolytica NCIM 3589 (761 MNNG treated and 39 MNNG + cerulenin treated). A three-stage screening using Sudan Black B plate technique, Nile red fluorimetry and total lipid extraction using solvent was performed, which enabled selection of ten high lipid yielding mutants. Time course studies of all the ten mutants were further undertaken in terms of biomass, lipid yield and lipid content to select three stable mutants (YlB6, YlC7 and YlE1) capable of growing and accumulating lipid on WCO, with lipid contents of 55, 60 and 67% as compared to 45% for the wild type. The mutants demonstrated increased volumetric lipid productivities (0.062, 0.044 and 0.041 g L−1 h−1) as compared to the wild type (0.033 g L−1 h−1). The fatty acid profile of the three mutants consisted of a high content of C16 and C18 saturated and monounsaturated fatty acids and was found to be suitable for biodiesel production. The fuel properties, namely, density, kinematic viscosity, total acid number, iodine value of the three mutants were evaluated and found to lie within the limits specified by internationally accepted standards. Additionally, it was noted that the mutants demonstrated better cetane numbers and higher heating values than the wild type strain.ConclusionThe chemical mutagenesis strategy adopted in this study resulted in the successful isolation of three stable high SCO yielding mutants. The mutants, namely, YlB6, YlC7 and YlE1 exhibited a 1.22, 1.33 and 1.49-fold increase in lipid contents when grown on 100 g L−1 waste cooking oil than the parental yeast strain. The fatty acid methyl ester (FAME) profiles of all the three mutants was determined to be suitable for biodiesel suggesting their potential applicability while simultaneously addressing the management of waste cooking oil.

Aberrant lipid metabolism in cancer cells - the role of oncolipid-activated signaling.

Metabolic activity of malignant cells is very different from that of their nontransformed equivalents, which establishes metabolic reprogramming as an important hallmark of every transformed cell. In particular, the current arena of research in this field aims to understand the regulatory effect of oncogenic signaling on metabolic rewiring in transformed cells in order to exploit this for therapeutic benefit. Alterations in lipid metabolism are one of the main aspects of metabolic rewiring of transformed cells. Up-regulation ofseveral lipogenic enzymes has been reported to be a characteristic of various cancer types. Lysophosphatidic acid (LPA), a simple byproduct of the lipid biosynthesis pathway, has gained immense importance due to its elevated level in several cancers and associated growth-promoting activity. Importantly, a current study revealed its role in increased denovo lipid synthesis through up-regulation of sterol regulatory element-binding protein 1, a master regulator of lipid metabolism. This review summarizes the recent insights in the field of oncolipid LPA-mediated signaling in regard to lipid metabolism in cancers. Future work in this domain is required to understand the up-regulation of the denovo synthesis pathway and the role of its end products in malignant cells. This will open a new arena of research toward the development of specific metabolic inhibitors that can add to the pre-existing chemotherapeutics in order to increase the efficacy of clinical output in cancer patients.

Scopoletin Supplementation Ameliorates Steatosis and Inflammation in Diabetic Mice.

Scopoletin is a bioactive component in many edible plants and fruits. This study investigated the effects of scopoletin on hepatic steatosis and inflammation in a high-fat diet fed type 1 diabetic mice by comparison with metformin. Scopoletin (0.01%, w/w) or metformin (0.5%, w/w) was provided with a high-fat diet to streptozotocin-induced diabetic mice for 11weeks. Both scopoletin and metformin lowered blood glucose and HbA1c , serum ALT, TNF-α and IL-6 levels, glucose intolerance, and hepatic lipid accumulation compared with the diabetic control group. Scopoletin or metformin down-regulated hepatic gene expression of triglyceride (Pparg, Plpp2, and Dgat2) and cholesterol (Hmgcr) synthesis as well as inflammation (Tlr4, Myd88, Nfkb1, Tnfa, and Il6), while it up-regulated Cyp7a1 gene. Hepatic PPARγ and DGAT2 protein levels were also down-regulated in scopoletin or metformin group compared with the control group. Scopoletin or metformin also inhibited hepatic fatty acid synthase and phosphatidate phosphohydrolase activities. These results suggest that scopoletin protects against diabetes-induced steatosis and inflammation by inhibiting lipid biosynthesis and TLR4-MyD88 pathways. Copyright © 2017 John Wiley & Sons, Ltd.

CgMED3 Changes Membrane Sterol Composition To Help Candida glabrata Tolerate Low-pH Stress.

Candida glabrata is a promising microorganism for organic acid production. The present study aimed to investigate the role of C. glabrata Mediator complex subunit 3 (CgMed3p) in protecting C. glabrata under low-pH conditions. To this end, genes CgMED3A and CgMED3B were deleted, resulting in the double-deletion Cgmed3ABΔ strain. The final biomass and cell viability levels of Cgmed3ABΔ decreased by 64.5% and 35.8%, respectively, compared to the wild-type strain results at pH 2.0. In addition, lack of CgMed3ABp resulted in selective repression of a subset of genes in the lipid biosynthesis and metabolism pathways. Furthermore, C18:1, lanosterol, zymosterol, fecosterol, and ergosterol were 13.2%, 80.4%, 40.4%, 78.1%, and 70.4% less abundant, respectively, in the Cgmed3ABΔ strain. In contrast, the concentration of squalene increased by about 44.6-fold. As a result, membrane integrity, rigidity, and H+-ATPase activity in the Cgmed3ABΔ strain were reduced by 62.7%, 13.0%, and 50.3%, respectively. In contrast, overexpression of CgMED3AB increased the levels of C18:0, C18:1, and ergosterol by 113.2%, 5.9%, and 26.4%, respectively. Moreover, compared to the wild-type results, dry cell weight and pyruvate production increased, irrespective of pH buffering. These results suggest that CgMED3AB regulates membrane composition, which in turn enables cells to tolerate low-pH stress. We propose that regulation of CgMed3ABp may provide a novel strategy for enhancing low-pH tolerance and increasing organic acid production by C. glabrataIMPORTANCE The objective of this study was to investigate the role of Candida glabrata Mediator complex subunit 3 (CgMed3ABp) and its regulation of gene expression at low pH in C. glabrata We found that CgMed3ABp was critical for cellular survival and pyruvate production during low-pH stress. Measures of the levels of plasma membrane fatty acids and sterol composition indicated that CgMed3ABp could play an important role in regulating homeostasis in C. glabrata We propose that controlling membrane lipid composition may enhance the robustness of C. glabrata for the production of organic acids.

Proteomic analysis of naturally occurring boron tolerant plant Gypsophila sphaerocephala L. in response to high boron concentration

Gypsophila sphaerocephala is a naturally Boron (B) tolerant species that can grow around the B mining areas in Turkey, where the B concentration in soil reaches a lethal dose for plants (up to ∼8900mgkg−1 (∼140mM). While its interesting survival capacity in extremely B containing soils, any molecular research has been conducted to understand its high tolerance mechanism yet. In the present study, we have performed a proteomic analysis of this plant to understand its high tolerance towards B-stress. Seedlings of G. sphaerocephala were collected from B mining area and were adapted to greenhouse conditions. An excessive level of Boric acid (3mM)was applied to the plantlets for 24h. Total proteins were precipitated by using TCA/Acetone method. 2D-PAGE (two-dimensional polyacrylamide gel electrophoresis) analysis of the proteins was carried out. Out of 121 protein spots, 14 were differentially expressed between the control and B-exposed G. sphaerocephala roots. The peptide profile of each protein was determined by MALDI-TOF mass spectrometer after in-gel trypsin digestion. The identified proteins are involved in different mechanisms in the cell such as in antioxidant mechanism, energy metabolism, protein degradation, lipid biosynthesis and signaling pathways, indicating that G. sphaerocephala has multiple cooperating mechanisms to protect itself from high B levels. Overall, this study sheds light on to the possible regulatory switches (gene/s) controlling the B-tolerance proteins and their possible roles in plant’s defense mechanism.

Abstract 2507: Using 1H NMR metabolomics to study breast cancer and endothelial cell metabolic interactions

Abstract Breast cancer is the second most commonly diagnosed cancer among women worldwide. Patient death is typically caused by metastasis development rather than the primary tumor. Metastasis in breast cancer has been shown to occur via blood and lymphatic vessels. Research shows that breast cancer cells ‘educate’ lymphatic and blood endothelial cells to support tumor growth by stimulating growth factor secretion; specifically VEGF, PDGF-BB and EGF secretion. In addition, cell metabolism is altered during malignant transformation. Cancer cells have increased energy and macromolecule biosynthesis requirements to sustain rapid proliferation. Increased angiogenesis observed in tumors points to a need for an increased nutrient supply. However, it remains unclear how tumor endothelial cell metabolism is altered and how metabolism regulates tumor angiogenesis. Therefore, this study aims to use 1H NMR metabolomics to identify breast cancer-endothelial cell metabolic interactions in a high-throughput manner. Firstly, we studied if in vitro co-culture with breast cancer altered endothelial cell metabolism. We identified distinct metabolic profiles for HUVECs grown in monoculture or co-culture with three different breast cancer cell lines. Metabolism of HUVECs co-cultured with MCF7 and SKBR3 breast cancer cell lines was significantly altered compared to mono-culture control. The most impacted metabolic pathways by co-culture were amino acid metabolism, energy metabolism and lipid biosynthesis pathways. Culturing HUVECs with MDA-MB-231 triple negative breast cancer cells did not have a significant impact on HUVEC metabolism, however. Together, these results indicate that culturing HUVECs with certain breast cancer lines causes significant changes in endothelial metabolism. Alterations in endothelial cell metabolism in response to breast cancer co-culture can further our understanding of tumor-vascular interactions and may lead to identification of metabolic biomarkers or therapeutic targets that can disrupt tumor angiogenesis. Citation Format: Suehelay Acevedo-Acevedo, Sean P. Palecek. Using 1H NMR metabolomics to study breast cancer and endothelial cell metabolic interactions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2507. doi:10.1158/1538-7445.AM2017-2507

ACCase and rbcL gene expression as a function of nutrient and metal stress for enhancing lipid productivity in Chlorella sorokiniana

Development of novel and scalable strategies for improving lipid and biomass yield is a prerequisite for sustainable biodiesel production from microalgae. Currently, nutrient stress is the widely employed lipid enhancement strategy in microalgae, yet, it is associated with compromised biomass productivity. In this study, an alternative approach for lipid enhancement in Chlorella sorokiniana is proposed to alleviate the constraint of low biomass production under nutrient stress. This is achieved through combining nutrient stress along with selected metals and EDTA stress. A substantial increase in lipid yield (77.03mgL−1d−1) was achieved when the nutrient (nitrogen and phosphorus) stress BG11 media was supplemented with a combination of iron (15mgL−1), magnesium (125mgL−1), calcium (18mgL−1) and EDTA (3mgL−1). This was further validated by quantifying the expression levels of two key genes involved in photosynthesis (rbcL) and lipid biosynthesis (accD) pathways. Under this developed strategy a 5.15-fold increase in rbcL and 9.79-fold increases in accD gene expression were noted in comparison to the culture grown in BG11 medium. A significant correlation could also be drawn between the expression of rbcL and accD genes to biomass yields, photosynthetic performance and lipid productivity. The proposed strategy could be easily applied at various commercial scale microalgal cultivation systems for enhancing lipid content without compromising the biomass yield.

Comparative transcriptome analysis of lipid biosynthesis in seeds and non-seed tissues of sea buckthorn

Sea buckthorn (Hippophae L.) is a woody crop with an unusual ability to thrive in very dry, nutrient-poor habitats. Seeds and berry pulp are rich in bioactive compounds with powerful medicinal properties, and are being used worldwide to an increasing extent. However, genomic information is still lacking for this crop, which limits the possibilities of raise the oil quality and yield with advanced breeding program. In this study, (1) sea buckthorn de novo transcriptome libraries for seeds, pulp, leaf, stem and root tissues were first constructed simultaneously, and 62,166 unigenes were assembled. Sixty-six differential expression genes (e-value cutoff <1 × 10−30) were found to be involved in lipid biosynthesis pathway, (2) based on the comparative transcriptome data, we first revealed the differentially expressed genes of oil biosynthesis and accumulation in seeds and berry pulp, and the high unigene expression of source gene ‘GPD1’ (3.61) contributed to synthesis more G3P of TAG precursor in pulp, and the high expression of sink genes ‘DGAT1 and DGAT2’ (14.57 and 12.57, respectively) accelerated high TAG accumulation in pulp, compared to their expressions in seeds (0, 1.24 and 6.85, respectively). The oil content in pulp (36.90%) was also higher than that in seed (14.15%), (3) qRT-PCR verification results showed the expressions of GPD1, DGAT1 and DGAT2 genes (1.46, 0.93 and 2.13 fold change, respectively) in pulp were also higher than those in seeds (0.47, 0.21 and 1.02 fold change, respectively). Our results suggest that the high coordinated expression of source ‘GPD1’ and sink ‘DGAT1 and DGAT2’ genes resulted in strong oil biosynthesis and accumulation in the pulp with high-oil pulp, compared to its seed with low-oil seeds. These transcriptome data provide genomic information for identification of genetic regulation system for sea buckthorn, especially understanding lipid biosynthesis mechanism in seeds and pulp. The detection of these three source and sink genes of lipid biosynthesis provided scientific basis for improving low oil contents in seeds and pulp of sea buckthorn berries.

Knowns and unknowns of membrane lipid synthesis in streptomycetes

Bacteria belonging to the genus Streptomyces are among the most prolific producers of antibiotics. Research on cellular membrane biosynthesis and turnover is lagging behind in Streptomyces compared to related organisms like Mycobacterium tuberculosis. While natural products discovery in Streptomyces is evidently a priority in order to discover new antibiotics to combat the increase in antibiotic resistant pathogens, a better understanding of this cellular compartment should provide insights into the interplay between core and secondary metabolism. However, some of the pathways for membrane lipid biosynthesis are still incomplete. In addition, while it has become clear that remodelling of the membrane is necessary for coping with environmental stress and for morphological differentiation, the detailed mechanisms of these adaptations remain elusive. Here, we aim to provide a summary of what is known about the polar lipid composition in Streptomyces, the biosynthetic pathways of polar lipids, and to highlight current gaps in understanding function, dynamics and biosynthesis of these essential molecules.

Transcriptome and functional analysis reveals hybrid vigor for oil biosynthesis in oil palm

Oil palm is the most productive oil crop in the world and composes 36% of the world production. However, the molecular mechanisms of hybrids vigor (or heterosis) between Dura, Pisifera and their hybrid progeny Tenera has not yet been well understood. Here we compared the temporal and spatial compositions of lipids and transcriptomes for two oil yielding organs mesocarp and endosperm from Dura, Pisifera and Tenera. Multiple lipid biosynthesis pathways are highly enriched in all non-additive expression pattern in endosperm, while cytokinine biosynthesis and cell cycle pathways are highly enriched both in endosperm and mesocarp. Compared with parental palms, the high oil content in Tenera was associated with much higher transcript levels of EgWRI1, homolog of Arabidopsis thaliana WRINKLED1. Among 338 identified genes in lipid synthesis, 207 (61%) has been identified to contain the WRI1 specific binding AW motif. We further functionally identified EgWRI1-1, one of three EgWRI1 orthologs, by genetic complementation of the Arabidopsis wri1 mutant. Ectopic expression of EgWRI1-1 in plant produced dramatically increased seed mass and oil content, with oil profile changed. Our findings provide an explanation for EgWRI1 as an important gene contributing hybrid vigor in lipid biosynthesis in oil palm.

In silico discovery and in vitro activity of inhibitors against Mycobacterium tuberculosis 7,8-diaminopelargonic acid synthase (Mtb BioA).

Computer-aided drug discovery and development approaches such as virtual screening, molecular docking, and in silico drug property calculations have been utilized in this effort to discover new lead compounds against tuberculosis. The enzyme 7,8-diaminopelargonic acid aminotransferase (BioA) in Mycobacterium tuberculosis (Mtb), primarily involved in the lipid biosynthesis pathway, was chosen as the drug target due to the fact that humans are not capable of synthesizing biotin endogenously. The computational screening of 4.5 million compounds from the Enamine REAL database has ultimately yielded 45 high-scoring, high-affinity compounds with desirable in silico absorption, distribution, metabolism, excretion, and toxicity properties. Seventeen of the 45 compounds were subjected to bioactivity validation using the resazurin microtiter assay. Among the 4 actives, compound 7 ((Z)-N-(2-isopropoxyphenyl)-2-oxo-2-((3-(trifluoromethyl)cyclohexyl)amino)acetimidic acid) displayed inhibitory activity up to 83% at 10 μg/mL concentration against the growth of the Mtb H37Ra strain.

Efficient gene targeting in non-homologous end-joining-deficient Lipomyces starkeyi strains.

Microbial lipids are sustainable feedstock for the production of oleochemicals and biodiesel. Oleaginous yeasts have recently been proposed as alternative lipid producers to plants and animals to promote sustainability in the chemical and fuel industries. The oleaginous yeast Lipomyces starkeyi has great industrial potential as an excellent lipid producer. However, improvement of its lipid productivity is essential for the cost-effective production of oleochemicals and fuels. Genetic and metabolic engineering of L. starkeyi via gene manipulation techniques may result in improvements in lipid production and our understanding of the mechanisms behind lipid biosynthesis pathways. We previously described an integrative transformation system using a drug-resistant marker for L. starkeyi. However, gene-targeting frequencies were very low because non-homologous recombination is probably predominant in L. starkeyi. Genetic engineering tools for L. starkeyi have not been sufficiently developed. In this study, we describe a new genetic tool and its application in L. starkeyi. To develop a highly efficient gene-targeting system for L. starkeyi, we constructed a series of mutants by disrupting genes for LsKu70p, LsKu80p, and/or LsLig4p, which share homology with other yeasts Ku70p, Ku80p, and Lig4p, respectively, being involved in non-homologous end-joining pathway. Deletion of the LsLIG4 gene dramatically improved the homologous recombination efficiency (80.0%) at the LsURA3 locus compared with that in the wild-type strain (1.4%), when 2000-bp homologous flanking regions were used. The homologous recombination efficiencies of the double mutant ∆l sku70∆lslig4 and the triple mutant ∆lsku70∆lsku80∆lslig4 were also markedly enhanced. Therefore, the L. starkeyi ∆lslig4 background strains have promise as efficient recipient strains for genetic and metabolic engineering approaches in this yeast.

Advances in culture and genetic modification approaches to lipid biosynthesis for biofuel production and in silico analysis of enzymatic dominions in proteins related to lipid biosynthesis in algae

SUMMARYBiofuels have been shown to be a promising and highly attractive alternative for minimizing the use of fossil fuels, and microalgae have positioned themselves as potential candidates for production of lipids and other substances of commercial interest. We briefly review recent advances made in microalgae culture conditions and genetic manipulation for improving lipid yields for biofuel production – with both approaches showing similar yields of triacylglycerides, indicating that more work is required for improving lipid yield and accumulation in algae. Aiming at gaining knowledge of algae genetic manipulation and exploring future use of this information for modifying the lipid biosynthesis pathway, we investigated whether some characteristics of enzymes involved in lipid biosynthesis could relate to lipid yield and accumulation in algae. We made an in silico analysis of amino acid sequence of enzymatic domains and modeled tertiary structure of three proteins involved in the biosynthesis of lipids in microalgae: acetyl‐CoA carboxylase, Acyl‐CoA: diacylglycerol acyltransferase, and glycerol‐3‐phosphate acyltransferase. Our results suggest that, based on primary amino acid sequences and tertiary structure of proteins shared by certain algae, it is likely that these proteins may relate to lipid yield and accumulation, which makes bioinformatics a powerful tool for in silico study of proteins and for selecting genes involved in lipid biosynthesis that could be useful for heterologous transformation in algae with the long term objective of improving their yield, accumulation, and fatty acid composition by genetic engineering.

Comparative genomics and expression profile of lipid biosynthesis pathway genes in Camellia sinensis

Comparative genomics and expression profile of lipid biosynthesis pathway genes in Camellia sinensis

RNAi-mediated down-regulation of the expression of OsFAD2-1: effect on lipid accumulation and expression of lipid biosynthetic genes in the rice grain.

BackgroundThe bran from polished rice grains can be used to produce rice bran oil (RBO). High oleic (HO) RBO has been generated previously through RNAi down-regulation of OsFAD2-1. HO-RBO has higher oxidative stability and could be directly used in the food industry without hydrogenation, and is hence free of trans fatty acids. However, relative to a classic oilseed, lipid metabolism in the rice grain is poorly studied and the genetic alteration in the novel HO genotype remains unexplored.ResultsHere, we have undertaken further analysis of role of OsFAD2-1 in the developing rice grain. The use of Illumina-based NGS transcriptomics analysis of developing rice grain reveals that knockdown of Os-FAD2-1 gene expression was accompanied by the down regulation of the expression of a number of key genes in the lipid biosynthesis pathway in the HO rice line. A slightly higher level of oil accumulation was also observed in the HO-RBO.ConclusionProminent among the down regulated genes were those that coded for FatA, LACS, SAD2, SAD5, caleosin and steroleosin. It may be possible to further increase the oleic acid content in rice oil by altering the expression of the lipid biosynthetic genes that are affected in the HO line.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-016-0881-6) contains supplementary material, which is available to authorized users.

Oil accumulation in non-seed tissue: transcriptome analysis of Chinese tallow

Chinese tallow ( Triadica sebifera ) is an oil-seed producing tree with a great potential as a biofuel crop and does not compete with food crops for land. The fruits are unique in that they contain two kinds of lipid, a highly un-saturated oil and a saturated fat separated in the seed and an external tallow layer respectively. However, until recently it was unclear whether the tallow is a non-seed tissue or an external fat deposition. We have addressed this important question by scanning electron microscopy to show that it is a cellular tissue and provided biochemical evidence for an active storage oil or triacylglycerol (TAG) biosynthetic machinery in the non-seed tallow tissue. We also showed that tallow tissue accumulates more TAG and has a different fatty acid profile compared to the seed TAG. As a sequence resource to aid the development of this oil crop, we generated a transcriptome resource by sequencing three component fruit tissues of tallow, seed and fruit coat. This enabled the reconstruction of lipid biosynthesis pathway in this potential biofuel crop. The transcriptome resource has thus far allowed identification of key differences in oil synthesis and storage mechanisms between the seed and tallow tissues. This non-seed oil accumulating tissue can be used as model system to gain further knowledge of such tissues to facilitate efficient utilization of non-seed biomass for oil production.

Highly conserved nucleotide phosphatase essential for membrane lipid homeostasis in Streptococcus pneumoniae.

Proteins belonging to the DHH family, a member of the phosphoesterase superfamily, are produced by most bacterial species. While some of these proteins are well studied in Bacillus subtilis and Escherichia coli, their functions in Streptococcus pneumoniae remain unclear. Recently, the highly conserved DHH subfamily 1 protein PapP (SP1298) has been reported to play an important role in virulence. Here, we provide a plausible explanation for the attenuated virulence of the papP mutant. Recombinant PapP specifically hydrolyzed nucleotides 3'-phosphoadenosine-5'-phosphate (pAp) and 5'-phosphoadenylyl-(3'->5')-adenosine (pApA). Deletion of papP, potentially leading to pAp/pApA accumulation, resulted in morphological defects and mis-localization of several cell division proteins. Incubation with both polar solvent and detergent led to robust killing of the papP mutant, indicating that membrane integrity is strongly affected. This is in line with previous studies showing that pAp inhibits the ACP synthase, an essential enzyme involved in lipid precursor production. Remarkably, partial inactivation of the lipid biosynthesis pathway, by inhibition of FabF or depletion of FabH, phenocopied the papP mutant. We conclude that pAp and pApA phosphatase activity of PapP is required for maintenance of membrane lipid homeostasis providing an explanation how inactivation of this protein may attenuate pneumococcal virulence.

Acylcarnitines participate in developmental processes associated to lipid metabolism in plants.

Plant acylcarnitines are present during anabolic processes of lipid metabolism. Their low contents relatively to the corresponding acyl-CoAs suggest that they are associated to specific pools of activated fatty acids. The non-proteinaceous amino acid carnitine exists in plants either as a free form or esterified to fatty acids. To clarify the biological significance of acylcarnitines in plant lipid metabolism, we have analyzed their content in plant extracts using an optimized tandem mass spectrometry coupled to liquid chromatography method. We have studied different developmental processes (post-germination, organogenesis, embryogenesis) targeted for their high requirement for lipid metabolism. The modulation of the acylcarnitine content was compared to that of the lipid composition and lipid biosynthetic gene expression level in the analyzed materials. Arabidopsis mutants were also studied based on their alteration in de novo fatty acid partitioning between the prokaryotic and eukaryotic pathways of lipid biosynthesis. We show that acylcarnitines cannot specifically be associated to triacylglycerol catabolism but that they are also associated to anabolic pathways of lipid metabolism. They are present during membrane and storage lipid biosynthesis processes. A great divergence in the relative contents of acylcarnitines as compared to the corresponding acyl-CoAs suggests that acylcarnitines are associated to very specific process(es) of lipid metabolism. The nature of their involvement as the transport form of activated fatty acids or in connection with the management of acyl-CoA pools is discussed. Also, the occurrence of medium-chain entities suggests that acylcarnitines are associated with additional lipid processes such as protein acylation for instance. This work strengthens the understanding of the role of acylcarnitines in plant lipid metabolism, probably in the management of specific acyl-CoA pools.