Tetraether Lipids Research Articles

Constraining the sources of archaeal tetraether lipids in multiple cold seep provinces of the Cascadia Margin

Archaeal isoprenoid glycerol dialkyl glycerol tetraether lipid (iGDGT) abundance profiles and carbon isotopic compositions reflect the relative distributions of archaeal sources, including planktonic, benthic, and methane-cycling contributions. Here, we analyze the carbon isotope ratios of iGDGTs purified from sediments of three different cold seep sites in Cascadia Margin, off the coast of Washington, USA. Together with relative abundance and glycerol configurations, we use the carbon isotope ratios to estimate the contributions of multiple archaeal sources to the sedimentary iGDGT assemblages and their impact on values of the TEX86 and methane indices. Using a Bayesian mixing model, we robustly characterize three potential endmembers by determining their characteristic lipid distributions, inferred contributions to the total sediment inventory, and carbon isotopic signatures. Despite the geographic proximity of the sample locations, we find site-specific heterogeneity in relative iGDGT abundances and δ13C values. Planktonic and benthic methane-cycling sources predominate in all cases (contributing > 98 % of iGDGTs), while benthic non-methane cycling archaea contribute minimally to the sedimentary lipid pool. Environments with higher methane influence show an increased presence of anti-parallel iGDGTs, indicating that methane-cycling archaea may dominantly or exclusively synthesize iGDGTs in this configuration. Our results quantify the relationship between the methane index (MI) and methane impact in systems dominated by planktonic and benthic methane-cycling archaea. Within the framework of the TEX86 temperature proxy, this permits a quantitative demonstration that it is overly simplistic to apply a MI cutoff threshold as a binary indicator to determine methane influence, and caution is needed when taking this approach in paleoclimate reconstructions.

Metabolic imprints in the hydrogen isotopes of Archaeoglobus fulgidus tetraether lipids

The stable hydrogen isotope composition of archaeal lipids is emerging as a potential paleoenvironmental proxy, adding to the well-established application of plant leaf wax-derived n-alkanes in paleohydrological reconstruction. A handful of studies reported relatively invariant and depleted hydrogen isotope compositions for archaeal lipids despite the range of different organisms and growth conditions explored. However, how modes of metabolism and physiological state (growth phase) affect the hydrogen isotope signatures of archaeal lipids remains poorly understood, limiting our ability to interpret archaeal lipid biomarker records from the environment. Here we conducted water isotope label experiments with a metabolically flexible and well-studied model archaeon Archaeoglobus fulgidus and quantified the hydrogen isotope fractionation between lipids and water in response to different carbon substrates and electron donor–acceptor pairs at different growth phases. The 2H/1H fractionation between lipids and water (εL/W) was overall negative. Both carbon metabolism and growth phase affected the magnitude of isotope fractionation in A. fulgidus; however, the changes in εL/W values were relatively subtle where they ranged from –283 to –229 ‰ across all tested conditions, overlapping with the ranges observed for other archaea in previous studies. Isotope flux-balance model results suggest that ≥ 80 % and ≥ 50 % of lipid-bound H in A. fulgidus cultures directly reflect water isotope compositions (i.e., not via organic substrate or H2) during autotrophy and heterotrophy, respectively. The model results also suggest two main mechanisms of consistent 2H depletion observed in A. fulgidus tetraethers as well as other archaeal lipids reported in previous studies: 1) isotopic re-equilibration via upstream isomerization reactions involving C5 units and 2) downstream double bond reduction catalyzed by a flavoenzyme geranylgeranyl reductase. These results are consistent with previous isotope flux-balance model results for a different archaeon. Finally, we synthesized available data to compare εL/W patterns across all three domains of life: Eukarya, Archaea and Bacteria. Because they vary fundamentally in lipid biosynthesis pathways, we present comparative discussions in pairs, focusing on the shared biochemical mechanisms among isoprenoid lipids and potential signals of metabolic adaptations across prokaryotic lipids. Emerging patterns between diverse archaeal and eukaryotic isoprenoid lipids are consistent with the two proposed mechanisms for 2H depletion identified (isomerization and final saturation). The patterns between archaeal isoprenoids and bacterial fatty acids suggest that the general state of energy limitation may also contribute to large, negative values of εL/W observed in prokaryotic lipids. Altogether, these findings lend further support for the potential of archaeal lipid εL/W as a paleohydrological proxy and provide a broader insight into the 2H/1H fractionation mechanisms potentially shared among prokaryotic and eukaryotic lipid biomarkers.

Oxyplasma meridianum gen. nov., sp. nov., an extremely acidophilic organotrophic member of the order Thermoplasmatales.

A mesophilic, hyperacidophilic archaeon, strain M1T, was isolated from a rock sample from Vulcano Island, Italy. Cells of this organism were cocci with an average diameter of 1 µm. Some cells possessed filaments. The strain grew in the range of temperatures between 15 and 52 °C and pH 0.5-4.0 with growth optima at 40 °C and pH 1.0. Strain M1T was aerobic and chemoorganotrophic, growing on complex substrates, such as casamino acids, trypticase, tryptone, yeast and beef extracts. No growth at expenses of oxidation of elemental sulphur or reduced sulphur compounds, pyrite, or ferrous sulphate was observed. The core lipids were glycerol dibiphytanyl glycerol tetraether lipids (membrane spanning) with 0 to 4 cyclopentane moieties and archaeol, with trace amounts of hydroxy archaeol. The dominant quinone was MK-7 : 7. The genome size of M1T was 1.67 Mbp with a G+C content of 39.76 mol%, and both characteristics were well within the common range for Thermoplasmatales. The phylogenetic analysis based on 16S rRNA gene sequence placed the strain M1T within the order Thermoplasmatales with sequence identities of 90.9, 90.3 and 90.5% to the closest SSU rRNA gene sequences from organisms with validly published names, Thermoplasma acidophilum, Thermoplasma volcanium and Thermogymnomonas acidicola, respectively. Based on the results of our genomic, phylogenetic, physiological and chemotaxonomic studies, we propose that strain M1T (=DSM 116605T=JCM 36570T) represents a new genus and species, Oxyplasma meridianum gen. nov., sp. nov., within the order Thermoplasmatales.

Effects of producing high levels of hyperthermophile-specific C25,C25-archaeal membrane lipids in Escherichia coli

A hyperthermophilic archaeon, Aeropyrum pernix, synthesizes C25,C25-archaeal membrane lipids, or extended archaeal membrane lipids, which contain two C25 isoprenoid chains that are linked to glycerol-1-phosphate via ether bonds and are longer than the usual C20,C20-archaeal membrane lipids. The C25,C25-archaeal membrane lipids are believed to allow the archaeon to survive under harsh conditions, because they are able to form lipid membranes that are impermeable at temperatures approaching the boiling point. The effect that C25,C25-archaeal membrane lipids exert on living cells, however, remains unproven along with an explanation for why the hyperthermophilic archaeon synthesizes these specific lipids instead of the more common C20,C20-archaeal lipids or double-headed tetraether lipids. To shed light on the effects that these hyperthermophile-specific membrane lipids exert on living cells, we have constructed an E. coli strain that produces C25,C25-archaeal membrane lipids. However, a resultant low level of productivity would not allow us to assess the effects of their production in E. coli cells. Herein, we report an enhancement of the productivity of C25,C25-archaeal membrane lipids in engineered E. coli strains via the introduction of metabolic pathways such as an artificial isoprenol utilization pathway where the precursors of isoprenoids are synthesized via a two-step phosphorylation of prenol and isoprenol supplemented to a growth medium. In the strain with the highest titer, a major component of C25,C25-archaeal membrane lipids reached ∼11 % of total lipids of E. coli. It is noteworthy that the high production of the extended archaeal lipids did not significantly affect the growth of the bacterial cells. The permeability of the cell membrane of the strain became slightly lower in the presence of the exogenous membrane lipids with longer hydrocarbon chains, which demonstrated the possibility to enhance bacterial cell membranes by the hyperthermophile-specific lipids, along with the surprising robustness of the E. coli cell membrane.

Redox indication potential of isoprenoid tetraether lipids in marine environments: Insights from the East China Sea

The response of marine ammonia-oxidizing archaea (AOA) to environmental stress is reflected in changes in their membrane lipid composition, particularly the unique isoprenoid glycerol dialkyl glycerol tetraethers (iGDGTs). However, the influence of dissolved oxygen (DO) on the composition of iGDGTs in the ocean remains unclear. This study aims to investigate the link between DO levels and the fractional abundances of iGDGTs in the East China Sea (ECS) to establish a redox proxy. Results suggested that the absolute abundances of iGDGTs were influenced by Thaumarchaeota biomass when DO concentrations exceeded 2 mg/L. Vertical distributions of iGDGTs through the water column suggested their transport from bottom waters to sediments. Increasing proportions of iGDGT-0 and the sum of iGDGT-1, iGDGT-2 and iGDGT-3 from suspended particulate matters to surface sediments indicated their preferential preservation. DO concentrations (2–6 mg/L) in the water column showed a significant positive correlation with the relative abundance of crenarchaeol (cren%) but a negative correlation with iGDGT-0%, suggesting insufficient DO levels to promote AOA cyclization. However, bottom DO concentrations exhibited significant negative correlations with both cren% and iGDGT-0% in surface sediments, attributed to enhanced cyclization of iGDGTs and Euryarchaeota abundance within more reducing sediments, respectively. The consistent relationship between iGDGT-0% and DO in both water column and sediments enabled iGDGT-0% to be a potential redox proxy. Temporal variations in iGDGT-0% in the ECS in recent decades aligned well with in-situ DO monitoring data, further validating iGDGT-0% as a promising redox proxy.

Archaeosomes for Oral Drug Delivery: From Continuous Microfluidics Production to Powdered Formulations.

Archaeosomes were manufactured from natural archaeal lipids by a microfluidics-assisted single-step production method utilizing a mixture of di- and tetraether lipids extracted from Sulfolobus acidocaldarius. The primary aim of this study was to investigate the exceptional stability of archaeosomes as potential carriers for oral drug delivery, with a focus on powdered formulations. The archaeosomes were negatively charged with a size of approximately 100 nm and a low polydispersity index. To assess their suitability for oral delivery, the archaeosomes were loaded with two model drugs: calcein, a fluorescent compound, and insulin, a peptide hormone. The archaeosomes demonstrated high stability in simulated intestinal fluids, with only 5% of the encapsulated compounds being released after 24 h, regardless of the presence of degrading enzymes or extremely acidic pH values such as those found in the stomach. In a co-culture cell model system mimicking the intestinal barrier, the archaeosomes showed strong adhesion to the cell membranes, facilitating a slow release of contents. The archaeosomes were loaded with insulin in a single-step procedure achieving an encapsulation efficiency of approximately 35%. These particles have been exposed to extreme manufacturing temperatures during freeze-drying and spray-drying processes, demonstrating remarkable resilience under these harsh conditions. The fabrication of stable dry powder formulations of archaeosomes represents a promising advancement toward the development of solid dosage forms for oral delivery of biological drugs.

Millennial changes and cooling trends in land surface warm-season temperatures during the Holocene

The scarcity of proxies and calibration models for quantitatively reconstructing millennial timescale seasonal temperature tremendously constraints our understanding of the Holocene thermal variation and its driven mechanisms. Here, we established two global warm-season temperature models by applying deep learning neural network analysis to the branched tetraether membrane lipids originating from surface soil and lacustrine sediment bacteria. We utilized these optimal models in global well-dated lacustrine, peatland, and loess profiles covering the Holocene. All reconstructions of warm-season temperatures, consistent with climate model simulations, indicate cooling trends since the early Holocene, primarily induced by decreased solar radiation in the Northern Hemisphere due to the precession peak at the early. We further demonstrated that the membrane lipids can effectively enhance the future millennial seasonal temperature research, including winter temperatures, without being restricted by geographical location and sedimentary carrier.

Distributions of bacterial branched tetraether lipids associated with bacterial communities in hummock and hollow soils from Southern Tibetan peatland

Branched glycerol dialkyl glycerol tetraethers (brGDGTs), special membrane-spanning lipids derived from some bacteria, are widely used as molecular tools in paleoenvironmental studies to reconstruct terrestrial temperature and pH conditions. However, the impact of microtopography discrepancies on the accuracy and applicability of brGDGT-based proxies remains unclear. Here we investigated the distributional patterns of brGDGTs and bacterial communities within the hummock-hollow microtopography in the mid-southern Tibetan peatlands. Our results show that the methylation index of C5-methyl brGDGTs (MBT′5ME) values are significantly correlated with the local mean annual air temperature (MAAT). However, the peat-specific temperature calibration exhibits an underestimation of MAATpeat compared to the present MAAT. We find the MBT′5ME values are significantly lower in the moist hollows than that in the drier hummocks. This discrepancy could be primarily attributed to the shifts in bacterial communities and the physiological adaptation of brGDGT-producing bacteria driven by the high soil water content. These findings highlight the remarkable variability of MBT′5ME values at small spatial scales, which holds profound implications for paleotemperature reconstruction in alpine peatlands.

Strong linkage between benthic oxygen uptake and bacterial tetraether lipids in deep-sea trench regions

Oxygen in marine sediments regulates many key biogeochemical processes, playing a crucial role in shaping Earth’s climate and benthic ecosystems. In this context, branched glycerol dialkyl glycerol tetraethers (brGDGTs), essential biomarkers in paleoenvironmental research, exhibit an as-yet-unresolved association with sediment oxygen conditions. Here, we investigated brGDGTs in sediments from three deep-sea regions (4045 to 10,100 m water depth) dominated by three respective trench systems and integrated the results with in situ oxygen microprofile data. Our results demonstrate robust correlations between diffusive oxygen uptake (DOU) obtained from microprofiles and brGDGT methylation and isomerization degrees, indicating their primary production within sediments and their strong linkage with microbial diagenetic activity. We establish a quantitative relationship between the Isomerization and Methylation index of Branched Tetraethers (IMBT) and DOU, suggesting its potential validity across deep-sea environments. Increased brGDGT methylation and isomerization likely enhance the fitness of source organisms in deep-sea habitats. Our study positions brGDGTs as a promising tool for quantifying benthic DOU in deep-sea settings, where DOU is a key metric for assessing sedimentary organic carbon degradation and microbial activity.

Distribution of branched glycerol dialkyl glycerol tetraether (brGDGT) lipids from soils and sediments from the same watershed are distinct regionally (central Chile) but not globally

Quantitative reconstructions of past continental climates are vital for understanding contemporary and past climate change. Branched glycerol dialkyl glycerol tetraethers (brGDGTs) are unique bacterial lipids that have been proposed as universal paleothermometers due to their correlation with temperature in modern settings. Thus, brGDGTs may serve as a crucial paleotemperature proxy for understanding past climate variations and improving regional climate projections, especially in critical but under constrained regions. That said, complications can arise in their application due to varying source contributions (e.g., soils vs. peats vs. lacustrine). As such, this study investigates brGDGT distributions in Chilean lake surface sediments and corresponding watershed soils to determine the source of brGDGTs to lake sediments. Global datasets of brGDGTs in lake sediments and soils were additionally compiled for comparison. Distinct brGDGT distributions in Chilean lakes and soils indicate minimal bias from soil inputs to the lacustrine sediments as well as in situ lacustrine production of brGDGTs, which supports the use of brGDGTs in lake sediments as reliable paleotemperature proxies in the region. The ΣIIIa/ΣIIa ratio, initially promising as a brGDGT source indicator in marine settings, shows global complexities in lacustrine settings, challenging the establishment of universal thresholds for source apportionment. That said, we show that the ratio can be successfully applied in Chilean lake surface sediments. Direct comparisons with watershed soils and further research are crucial for discerning brGDGT sources in lake sediments and improving paleotemperature reconstructions on regional and global scales moving forward. Overall, this study contributes valuable insights into brGDGT variability, essential for accurate paleoreconstructions.

Multi-proxy temperature and environmental reconstruction during the Late Glacial and Early Holocene in the Bohemian Forest, Central Europe

Multi-proxy temperature reconstructions can provide robust insights into past environmental conditions. By combining different proxies we can disentangle the temperature signal from the indirect climate effects on the environment. This study uses a multi-proxy approach to reconstruct temperature and palaeoenvironmental conditions during the Late Glacial and Early Holocene (13.5–8 cal. ka BP) in the Bohemian Forest, Central Europe. We assessed the similarity of the temperature signal based on chironomids, isoprenoid glycerol dialkyl glycerol tetraether lipids (isoGDGTs), and pollen within a comparison with locally modeled temperature data generated by the CHELSA_Trace21k dataset. Pollen, macroscopic charcoal remains, and geochemistry were further used to reconstruct past environmental conditions such as vegetation dynamics, fire activity, the input of lithogenic material (Titanium), nutrient content (Total Nitrogen) and the sources of organic matter (C/N and δ13Corg). All temperature reconstructions based on independent proxies were positively correlated and followed the same long-term trend. However, results also showed that chironomids-inferred July temperature had lower amplitude variations compared to the other temperature curves. IsoGDGTs showed the most pronounced decrease in temperature values at the onset of the Younger Dryas (YD), corroborating that this cooling event was more marked during winter than summer. However, a decrease of less than 1 °C during summer and two short-term warm events at 12.6 and 12.2 cal ka BP provoked a modest and asynchronous response of the vegetation to the onset of the YD. Nevertheless, isoGDGTs appeared to react to changes in both temperature and organic carbon sources, particularly between 11.2 and 10.6 cal yr BP. These environmental changes, characterized by high values of the GDGT-0/crenarchaeol ratio, recorded an increase in methanogenic activity in the lake sediments, which likely altered the recorded climatic signal. The corresponding anoxic episodes in the lake sediments might be caused by an increasing input of organic carbon from the catchment, related to the development of the vegetation and catchment soils at the beginning of the Holocene. Finally, pollen-based temperature reconstruction showed a lag in the response to major climatic events, such as the onset of YD and Holocene. Our study increases the understanding of the climate-vegetation-environmental feedback during the Late Glacial and Early Holocene in the Bohemian Forest, Central Europe.

Absence of a water depth signal in archaeal tetraether lipids from surface lake sediments in areas with intensive human activity: A case study from Daihai Lake, Inner Mongolia

Indices based on isoprenoid glycerol dialkyl glycerol tetraethers (isoGDGTs) and hydroxylated GDGTs (OH-GDGTs), such as %Cren, Cren/Cren', and %OH-GDGTs, have been widely used as lake-level proxies in lake sediment cores. However, the interpretation of these indices is potentially complex and more research is needed to determine the contexts in which they can reliably be used for paleohydrological reconstruction. We analyzed the isoGDGTs and OH-GDGTs of multiple sample types (watershed surface soils, lake surface sediments, and sediments from a 70-cm sediment core) from Daihai Lake. Our results show that while the widely-used lake level proxies are not correlated with water depth for the surface sediments, this correlation exists in the samples from the sediment core over the past 60 years. We propose that archaeal GDGTs of modern surface sediments in Daihai Lake may be modified by intensive human activities, including eutrophication and potential terrestrial soil input, which prevents their use for lake-level reconstruction. Data from three other lakes (Qinghai, Lugu, and Yangzong) confirm that lake level proxies based on surface sediments are applicable for Qinghai Lake and Lugu Lake (both of which are in regions with relatively weak human activity), but not for Yangzong Lake (in a region with intensive human activity). Interestingly, the transition zone between Qinghai Lake/Lugu Lake and Daihai Lake/Yangzong Lake corresponds to the Hu Line, which divides China into two parts with contrasting human population densities. We propose that future research should consider these findings when using surface lake sediments for paleoenvironmental calibration in areas with intensive human activity, and that sediment cores may sometimes be more reliable for modern validation studies. Although more studies are needed to verify our conclusions, our findings contribute to the application of archaeal GDGTs to lake-level reconstruction, and they are potentially also significant for the validation of other proxies.

Peatland warming influences the abundance and distribution of branched tetraether lipids: Implications for temperature reconstruction

Branched glycerol dialkyl glycerol tetraethers (brGDGTs) are bacterial membrane lipids whose distribution in peatland soils serves as an important proxy for past climate changes due to strong linear correlations with temperature in modern environments. However, commonly used brGDGT-based temperature models are characterized by high uncertainty (ca. 4 °C) and these calibrations can show implausible correlations when applied at an ecosystem level. This lack of accuracy is often attributed to our limited understanding of the exact mechanisms behind the relationship between brGDGTs and temperature and the potential effect of temperature-independent factors on brGDGT distribution. Here, we examine the abundance and distribution of brGDGTs in a boreal peatland after four years of in-situ warming (+0, +2.25, +4.5, +6.75 and +9 °C). We observed that with warming, concentrations of total brGDGTs increased. Furthermore, we determined a shift in brGDGT distribution in the surface aerobic layers of the acrotelm (0–30 cm depth), whereas no detectable change was observed at deeper anaerobic depths (>40 cm), possibly due to limited microbial activity. The response of brGDGTs to warming was also reflected by a strong increase in the methylation index of 5-methyl brGDGTs (MBT′5Me), classically used as a temperature proxy. Further, the relationship between the MBT′5Me index and soil temperature differed between 0–10, 10–20 and 20–30 cm depth, highlighting depth-specific response of brGDGTs to warming, which should be considered in paleoenvironmental and paleoecological studies. As the bacterial community composition was generally unaltered, the rapid changes in brGDGT distribution argue for a physiological adaptation of the microorganisms producing these lipids. Finally, soil temperature and water table depth were better predictors of brGDGT concentration and distribution, highlighting the potential for these drivers to impact brGDGT-based proxies. To summarize, our results provide insights on the response of brGDGT source microorganisms to soil warming and underscore brGDGTs as viable temperature proxies for better understanding of climatic perturbation in peatlands.

In vitro and in ovo photodynamic efficacy of nebulized curcumin-loaded tetraether lipid liposomes prepared by DC as stable drug delivery system

Lung cancer is one of the most common causes of high mortality worldwide. Current treatment strategies, e.g., surgery, radiotherapy, chemotherapy, and immunotherapy, insufficiently affect the overall outcome. In this study, we used curcumin as a natural photosensitizer in photodynamic therapy and encapsulated it in liposomes consisting of stabilizing tetraether lipids aiming for a pulmonary drug delivery system against lung cancer. The liposomes with either hydrolyzed glycerol-dialkyl-glycerol tetraether (hGDGT) in different ratios or hydrolyzed glycerol-dialkyl-nonitol tetraether (hGDNT) were prepared by dual centrifugation (DC), an innovative method for liposome preparation. The liposomes’ physicochemical characteristics before and after nebulization and other nebulization characteristics confirmed their suitability. Morphological characterization using atomic force and transmission electron microscopy showed proper vesicular structures indicative of liposomes. Qualitative and quantitative uptake of the curcumin-loaded liposomes in lung adenocarcinoma (A549) cells was visualized and proven. Phototoxic effects of the liposomes were detected on A549 cells, showing decreased cell viability. The generation of reactive oxygen species required for PDT and disruption of mitochondrial membrane potential were confirmed. Moreover, the chorioallantoic membrane (CAM) model was used to further evaluate biocompatibility and photodynamic efficacy in a 3D cell culture context. Photodynamic efficacy was assessed by PET/CT after nebulization of the liposomes onto the xenografted tumors on the CAM with subsequent irradiation. The physicochemical properties and the efficacy of tetraether lipid liposomes encapsulating curcumin, especially liposomes containing hGDNT, in 2D and 3D cell cultures seem promising for future PDT usage against lung cancer.

Comprehensive molecular-isotopic characterization of archaeal lipids in the Black Sea water column and underlying sediments.

The Black Sea is a permanently anoxic, marine basin serving as model system for the deposition of organic-rich sediments in a highly stratified ocean. In such systems, archaeal lipids are widely used as paleoceanographic and biogeochemical proxies; however, the diverse planktonic and benthic sources as well as their potentially distinct diagenetic fate may complicate their application. To track the flux of archaeal lipids and to constrain their sources and turnover, we quantitatively examined the distributions and stable carbon isotopic compositions (δ13 C) of intact polar lipids (IPLs) and core lipids (CLs) from the upper oxic water column into the underlying sediments, reaching deposits from the last glacial. The distribution of IPLs responded more sensitively to the geochemical zonation than the CLs, with the latter being governed by the deposition from the chemocline. The isotopic composition of archaeal lipids indicates CLs and IPLs in the deep anoxic water column have negligible influence on the sedimentary pool. Archaeol substitutes tetraether lipids as the most abundant IPL in the deep anoxic water column and the lacustrine methanic zone. Its elevated IPL/CL ratios and negative δ13 C values indicate active methane metabolism. Sedimentary CL- and IPL-crenarchaeol were exclusively derived from the water column, as indicated by non-variable δ13 C values that are identical to those in the chemocline and by the low BIT (branched isoprenoid tetraether index). By contrast, insitu production accounts on average for 22% of the sedimentary IPL-GDGT-0 (glycerol dibiphytanyl glycerol tetraether) based on isotopic mass balance using the fermentation product lactate as an endmember for the dissolved substrate pool. Despite the structural similarity, glycosidic crenarchaeol appears to be more recalcitrant in comparison to its non-cycloalkylated counterpart GDGT-0, as indicated by its consistently higher IPL/CL ratio in sediments. The higher TEX86 , CCaT, and GDGT-2/-3 values in glacial sediments could plausibly result from selective turnover of archaeal lipids and/or an archaeal ecology shift during the transition from the glacial lacustrine to the Holocene marine setting. Our in-depth molecular-isotopic examination of archaeal core and intact polar lipids provided new constraints on the sources and fate of archaeal lipids and their applicability in paleoceanographic and biogeochemical studies.

Abstract 1983 Identifying the Role of the Rubredoxin-like Domain in Tetraether Lipid Synthesis

Abstract 1983 Identifying the Role of the Rubredoxin-like Domain in Tetraether Lipid Synthesis

Tetraether archaeal lipids promote long-term survival in extreme conditions.

The sole unifying feature of the incredibly diverse Archaea is their isoprenoid-based ether-linked lipid membranes. Unique lipid membrane composition, including an abundance of membrane-spanning tetraether lipids, impart resistance to extreme conditions. Many questions remain, however, regarding the synthesis and modification of tetraether lipids and how dynamic changes to archaeal lipid membrane composition support hyperthermophily. Tetraether membranes, termed glycerol dibiphytanyl glycerol tetraethers (GDGTs), are generated by tetraether synthase (Tes) by joining the tails of two bilayer lipids known as archaeol. GDGTs are often further specialized through the addition of cyclopentane rings by GDGT ring synthase (Grs). A positive correlation between relative GDGT abundance and entry into stationary phase growth has been observed, but the physiological impact of inhibiting GDGT synthesis has not previously been reported. Here, we demonstrate that the model hyperthermophile Thermococcus kodakarensis remains viable when Tes (TK2145) or Grs (TK0167) are deleted, permitting phenotypic and lipid analyses at different temperatures. The absence of cyclopentane rings in GDGTs does not impact growth in T. kodakarensis, but an overabundance of rings due to ectopic Grs expression is highly fitness negative at supra-optimal temperatures. In contrast, deletion of Tes resulted in the loss of all GDGTs, cyclization of archaeol, and loss of viability upon transition to the stationary phase in this model archaea. These results demonstrate the critical roles of highly specialized, dynamic, isoprenoid-based lipid membranes for archaeal survival at high temperatures.

Variations in isoprenoid tetraether lipids through the water column of the Western Pacific Ocean: Implications for sedimentary TEX86 records

The TetraEther indeX of 86 carbon atoms (TEX86) is widely used as a proxy to reconstruct past sea surface temperatures. Most current applications of TEX86 are primarily based on analyzing the composition of isoprenoid glycerol dialkyl glycerol tetraethers (isoGDGTs) that comprise TEX86 in sediments, with the assumption that the sedimentary isoGDGTs are mainly derived from the surface mixed layer. Here we report on the variations in the isoGDGT distribution, archaeal abundance and community through the water column of the Western Pacific Ocean, directly testing the export depth of isoGDGTs and constraining the temperature records of TEX86. Our data show that maximum isoGDGT concentrations occurred in subsurface waters (150–200 m) with maximum archaeal abundances. The ratio between isoGDGTs bearing 2 vs. 3 cyclopentane moieties, i.e. [2/3] ratio, increased with depth, which is likely related to the shift of the archaeal community from Ca. Nitrosopelagicus-dominance to norank_f__Nitrosopumilaceae-dominance. Models based on the [2/3] ratios in the water column predicted an average export depth of isoGDGTs to sediments of around 150–200 m, consistent with the robust relationship between the compiled sedimentary TEX86 and the annual mean subsurface temperature. Taken together, our findings support that TEX86 records subsurface rather than surface temperatures in the open ocean.

Archaea membranes in response to extreme acidic environments

Bipolar tetraether lipids (BTL), such as glycerol dialkyl calditol tetraether (GDNT) and glycerol dialkyl glycerol tetraether (GDGT), are the dominating lipid species in thermoacidophiles that inhabit at pH ≤ 4 and temperatures ≥65°C. BTL containing archaea membranes respond to environmental pH changes by varying the number of cyclopentane rings in the isoprenoids, the amount of GDNT relative to GDGT, the ratio of tetraethers to diethers, and the level of glycosylation in polar headgroups. These structural and compositional adjustments can alter the hydrogen bond networks in the membrane polar headgroup regions and the packing tightness and rigidity in the membrane hydrophobic core. It is likely that these changes in non-covalent interactions among archaea lipids are made to retain low membrane volume fluctuations and their low sensitivity to temperature, as illustrated in the case of liposomes made of the polar lipid fraction E (PLFE) of Sulfolobus acidocaldarius. As such, a low passive proton permeability and a near neutral intracellular pH can be maintained, and, as a result, optimal activities of soluble and membrane-bound proteins in thermoacidophiles can be retained in acidic growth conditions at elevated growth temperatures.

Structure-function relationships in pure archaeal bipolar tetraether lipids.

Archaeal bipolar tetraether lipids (BTLs) are among the most unusual lipids occurring in nature because of their presumed ability to span the entire membrane to form a monolayer structure. It is believed that because of their unique structural organization and chemical stability, BTLs offer extraordinary adaptation to archaea to thrive in the most extreme milieus. BTLs have also received considerable attention for development of novel membrane-based materials. Despite their fundamental biological significance and biotechnological interests, prior studies on pure BTLs are limited because of the difficulty to extract them in pure form from natural sources or to synthesize them chemically. Here we have utilized chemical synthesis to enable in-depth biophysical investigations on a series of chemically pure glycerol dialkyl glycerol tetraether (GDGT) lipids. The lipids self-assemble to form membrane-bound vesicles encapsulating polar molecules in aqueous media, and reconstitute a functional integral membrane protein. Structural properties of the membranes were characterized via small-angle X-ray scattering (SAXS) and cryogenic electron microscopy (cryo-EM). SAXS studies on bulk aqueous dispersions of GDGT lipids over 10-90 °C revealed lamellar and non-lamellar phases and their transitions. Next we asked whether vesicles overwhelmingly composed of a single GDGT species can undergo fusion as it is difficult to conceptualize such behavior with the assumption that such membranes have a monolayer structure. Interestingly, we observed that GDGT vesicles undergo fusion with influenza virus with lipid mixing kinetics comparable to that with vesicles composed of monopolar phospholipids. Our results suggest that GDGT membranes may consist of regions with a bilayer structure or form bilayer structures transiently which facilitate fusion and thus offer insight into how archaea may perform important physiological functions that require dynamical membrane behavior.