Long-chain Polyamines Research Articles

Preparation of thick silica coatings on carbon fibers with fine-structured silica nanotubes induced by a self-assembly process.

A facile method to coat carbon fibers with a silica shell is presented in this work. By immobilizing linear polyamines on the carbon fiber surface, the high catalytic activity of polyamines in the sol–gel-processing of silica precursors is used to deposit a silica coating directly on the fiber’s surface. The surface localization of the catalyst is achieved either by attaching short-chain polyamines (e.g., tetraethylenepentamine) via covalent bonds to the carbon fiber surface or by depositing long-chain polyamines (e.g., linear poly(ethylenimine)) on the carbon fiber by weak non-covalent bonding. The long-chain polyamine self-assembles onto the carbon fiber substrate in the form of nanoscopic crystallites, which serve as a template for the subsequent silica deposition. The silicification at close to neutral pH is spatially restricted to the localized polyamine and consequently to the fiber surface. In case of the linear poly(ethylenimine), silica shells of several micrometers in thickness can be obtained and their morphology is easily controlled by a considerable number of synthesis parameters. A unique feature is the hierarchical biomimetic structure of the silica coating which surrounds the embedded carbon fiber by fibrillar and interconnected silica fine-structures. The high surface area of the nanostructured composite fiber may be exploited for catalytic applications and adsorption purposes.

3D-Membrane Stacks on Supported Membranes Composed of Diatom Lipids Induced by Long-Chain Polyamines.

Long-chain polyamines (LCPAs) are intimately involved in the biomineralization process of diatoms taking place in silica deposition vesicles being acidic compartments surrounded by a lipid bilayer. Here, we addressed the question whether and how LCPAs interact with lipid membranes composed of glycerophospholipids and glyceroglycolipids mimicking the membranes of diatoms and higher plants. Solid supported lipid bilayers and monolayers containing the three major components that are unique in diatoms and higher plants, i.e., monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), and sulfoquinovosyldiacylglycerol (SQDG), were prepared by spreading small unilamellar vesicles. The integrity of the membranes was investigated by fluorescence microscopy and atomic force microscopy showing continuous flat bilayers and monolayers with small protrusions on top of the membrane. The addition of a synthetic polyamine composed of 13 amine groups separated by a propyl spacer (C3N13) results in flat but three-dimensional membrane stacks within minutes. The membrane stacks are connected with the underlying membrane as verified by fluorescence recovery after photobleaching experiments. Membrane stack formation was found to be independent of the lipid composition; i.e., neither glyceroglycolipids nor negatively charged lipids were required. However, the formation process was strongly dependent on the chain length of the polyamine. Whereas short polyamines such as the naturally occurring spermidine, spermine, and the synthetic polyamines C3N4 and C3N5 do not induce stack formation, those containing seven and more amine groups (C3N7, C3N13, and C3N18) do form membrane stacks. The observed stack formation might have implications for the stability and expansion of the silica deposition vesicle during valve and girdle band formation in diatoms.

Biosynthesis of polyamines and polyamine-containing molecules.

Polyamines are evolutionarily ancient polycations derived from amino acids and are pervasive in all domains of life. They are essential for cell growth and proliferation in eukaryotes and are essential, important or dispensable for growth in bacteria. Polyamines present a useful scaffold to attach other moieties to, and are often incorporated into specialized metabolism. Life has evolved multiple pathways to synthesize polyamines, and structural variants of polyamines have evolved in bacteria, archaea and eukaryotes. Among the complex biosynthetic diversity, patterns of evolutionary reiteration can be distinguished, revealing evolutionary recycling of particular protein folds and enzyme chassis. The same enzyme activities have evolved from multiple protein folds, suggesting an inevitability of evolution of polyamine biosynthesis. This review discusses the different biosynthetic strategies used in life to produce diamines, triamines, tetra-amines and branched and long-chain polyamines. It also discusses the enzymes that incorporate polyamines into specialized metabolites and attempts to place polyamine biosynthesis in an evolutionary context.

Synthesis and NMR analysis of 13C and 15N-labeled long-chain polyamines (LCPAs)

Site specific isotope labels in long-chain polyamines (LCPAs) for the NMR analysis of bioinspired silica nanocomposites.

Insight into the Supramolecular Architecture of Intact Diatom Biosilica from DNP-Supported Solid-State NMR Spectroscopy.

Diatom biosilica is an inorganic/organic hybrid with interesting properties. The molecular architecture of the organic material at the atomic and nanometer scale has so far remained unknown, in particular for intact biosilica. A DNP-supported ssNMR approach assisted by microscopy, MS, and MD simulations was applied to study the structural organization of intact biosilica. For the first time, the secondary structure elements of tightly biosilica-associated native proteins in diatom biosilica were characterized in situ. Our data suggest that these proteins are rich in a limited set of amino acids and adopt a mixture of random-coil and β-strand conformations. Furthermore, biosilica-associated long-chain polyamines and carbohydrates were characterized, thereby leading to a model for the supramolecular organization of intact biosilica.

Chemoselective silicification of synthetic peptides and polyamines.

Biosilicification sets the standard for the localized in vitro precipitation of silica at low orthosilicate concentrations in aqueous environment under ambient conditions. Numerous parameters must be controlled for the development of new technologies in designing inventive nanosilica structures, which are able to challenge the biological templates. A long neglected requirement that came into focus in the recent years are the cellular techniques of preventing unintentional lithification of cellular structures since numerous cellular components such as membranes, DNA, and proteins are known to precipitate nanosilica. The diatom metabolism makes use of techniques that restrict silicification to an armor of silica around the cell wall while avoiding the petrifying gaze of Medusa, which turns the whole cell into stone. Step by step, biochemistry unveils the hierarchical interplay of an arsenal of low-molecular weight molecules, proteins, and the cytoskeletal architecture and it becomes clearer why the organisms invest much metabolic effort for an obviously simple chemical reaction like the precipitation of amorphous silica. The discrimination between different soluble components in the silicification process (chemoselective silicification) is not only vitally important for the diatom but poses an interesting challenge for in vitro experiments. Until now, silica precipitation studies were mainly focused on the amount, the morphology, and composition of the precipitate while disregarding a quantitative analysis of the remaining soluble components. Here, we turn the tables and quantify the soluble components by 1H NMR in the progress of precipitation and present experiments which quantify the additivity, and potential cooperativity of long chain polyamines (LCPAs) and cationic peptides in the silicification process.

Electrostatic interplay: The interaction triangle of polyamines, silicic acid, and phosphate studied through turbidity measurements, silicomolybdic acid test, and 29Si NMR spectroscopy

The discovery of long-chain polyamines as biomolecules that are tightly associated to biosilica in diatom cell walls has inspired numerous in vitro studies aiming to characterize polyamine–silica interactions. The determination of these interactions at the molecular level is of fundamental interest on one hand for the understanding of cell wall biogenesis in diatoms and on the other hand for designing bioinspired materials synthesis approaches. The present contribution deals with the influence of amines and polyamines upon the initial self-assembly processes taking place during polyamine-mediated silica formation in solution. The influence of phosphate upon these processes is studied. For this purpose, sodium metasilicate solutions containing additives such as polyallylamine, allylamine and others in the presence/absence of phosphate were investigated. The analyses are based mainly on turbidity measurements yielding information about the early aggregation steps which finally give rise to the formation and precipitation of silica.

Assembly of multiple components in a hybrid microcapsule: designing a magnetically separable Pd catalyst for selective hydrogenation.

In analogy to the role of long-chain polyamines in biosilicification, poly-L-lysine facilitates the assembly of nanocomponents to design multifunctional microcapsule structures. The method is demonstrated by the fabrication of a magnetically separable catalyst that accommodates Pd nanoparticles (NPs) as active catalyst, Fe3O4 NPs as magnetic component for easy recovery of the catalyst, and silica NPs to impart stability and selectivity to the catalyst. In addition, polyamines embedded inside the microcapsule prevent the agglomeration of Pd NPs and thus result in efficient catalytic activity in hydrogenation reactions, and the hydrophilic silica surface results in selectivity in reactions depending on the polarity of substrates.

Enabling Antibacterial Coating via Bioinspired Mineralization of Nanostructured ZnO on Fabrics under Mild Conditions

Herein, we present an environmentally benign method capable of mineralization and deposition of nanomaterials to introduce antibacterial functionalities into cotton fabrics under mild conditions. Similar to the way in which many naturally occurring biominerals evolve around the living organism under ambient conditions, this technique enables flexible substrates like the cotton fabric to be coated with inorganic-based functional materials. Specifically, our strategy involves the use of long-chain polyamines known to be responsible in certain biomineralization processes, to nucleate, organize, and deposit nanostructured ZnO on cotton bandage in an aqueous solution under mild conditions of room temperature and neutral pH. The ZnO-coated cotton bandages as characterized by SEM, confocal micro-Raman spectroscopy, XRD, UV-DRS, and fluorescence microscopy demonstrate the importance of polyamine in generating a stable and uniform coating of spindle-shaped ZnO particles on individual threads of the fabric. As the coating process requires only mild conditions, it avoids any adverse effect on the thermal and mechanical properties of the substrate. Furthermore, the ZnO particles on cotton fabric show efficient antibacterial activity against both gram-positive and gram-negetive bacteria. Therefore, the developed polyamine mediated bioinspired coating method provides not only a facile and "green" synthesis for coating on flexible substrate but also the fabrication of antibacterial enabled materials for healthcare applications.

Diatom microfossils from cretaceous and eocene sediments contain native silica precipitating long‐chain polyamines

Organic molecules from known biological sources (biomarkers) that are preserved over geological time are critical tools in the study of past conditions and events on earth. Polar molecules are typically recycled rapidly in marine environments and do not survive burial within aquatic sediments in unambiguously recognizable form. As such, geological biomarkers are formed almost exclusively from precursor biomolecules that have been altered, limiting their utility as paleoproxies. Here, we report that nitrogen-rich aliphatic long-chain polyamines (LCPAs), biosynthesized by diatoms in species-specific assemblages for the precipitation of nanopatterned siliceous cell walls (frustules), are preserved unaltered in the oldest available diatom fossils dating to the Lower Cretaceous (early Albian, 115-110Ma). We further show that the cumulative LCPA pool accounts for 60% of the total C and 80% of the total N preserved in the Cretaceous age sediments. We suggest that silica glass formation by diatoms constitutes an important preservation mechanism for source-specific, polar biomolecules, protecting them indefinitely by encapsulation within the silicified frustule. LCPAs are a unique, source-specific carbon and nitrogen archive of diatom biomass, offering a promising tool for reconstruction of global cycles of carbon and nitrogen over geological timescales.

Controlled morphogenesis of amorphous silica and its relevance to biosilicification

Biogenetic biosilica displays intricate patterns that are structured on a nanometer-to-micrometer scale. At the nanoscale, it involves the polymerization products of silica, apparently mediated by the interaction between different biomolecules with special functional groups. In this paper, using tetraethyl orthosilicate [TEOS, Si(OCH2CH3)4] as a silica source, phospholipid (PL) and dodecylamine (DA) were introduced as model organic additives to investigate their influence on the formation and morphology of silica in the mineralization process. Morphology, structure, and composition of the products were characterized using a range of techniques including FESEM, TEM, SAXRD, TG-DTA, solid-state 29Si NMR, FTIR, and nitrogen physisorption. The FESEM and TEM analyses demonstrate that increasing PL concentrations at constant DA content leads to the formation of siliceous elongated structures. Localized enlargement can also be observed during further growth of elongated structures, displaying some features of the earliest recognizable stage of valve development in diatoms. In addition, in the presence or absence of PL, a series of control experiments using ammonia instead of DA show that no elongated structures are obtained, suggesting that the formation of elongated silica structures results from the cooperative interactions between PL and DA molecules. Because both organic amines (e.g., long-chain polyamines, LCPA) and phospholipid membranes (e.g., silicalemma) are of special importance for biosilicification in diatoms and sponges, our results imply that phospholipids are involved in the formation of organic aggregates, and thus influence the amines-mediated silica deposition. As such, our results may provide a new insight into the mechanism of biosilicification.

Widespread distribution and molecular diversity of diatom frustule bound aliphatic long chain polyamines (LCPAs) in marine sediments

The siliceous skeletal remains of diatoms are a primary component of sediments, covering vast areas of the sea floor. We used high performance liquid chromatography–electrospray ionization–tandem mass spectrometry (HPLC–ESI–MSn) under high resolution time-of-flight MS (TOF-MS) conditions, as well as nuclear magnetic resonance spectroscopy (NMR) and Fourier transform infrared spectroscopy (FTIR), to investigate the distribution and molecular diversity of long chain polyamines (LCPAs) entrapped in sedimentary diatom biosilica in sediments from the Indian sector of the Southern Ocean, the Bering Sea and the Northeast Pacific. This revealed the existence of complex polyamine populations with both known and unknown structural features, including the presence of ketone and carboxylic acid groups in the LCPA backbone, functional groups that likely modulate the silica precipitating properties of LCPAs in the environment and which may constitute points of initial silica deposition (Wallace, A.F., De Yoreso, J.J., Dove, P.M., 2009. Kinetics of silica nucleation on carboxy- and amine-terminated surfaces: insights for biomineralization. Journal of the American Chemical Society 131, 5244–5250). Although various LCPA species overlapped between samples (putrescine-based LCPAs with various degrees of methylation and N-methyl propylamine repeat units were ubiquitous in all samples), clear regional differences were shown in composition and structural characteristics, likely reflecting the various diatom species contributing to the total LCPA pool for each sedimentary environment. We further show, using scanning electron microscopy and energy dispersive X-ray analysis (SEM–EDX) that LCPAs purified from diatom skeletal remains rapidly re-direct the formation of silica nanospheres in vitro, and become embedded in the precipitated silicified matrix, forming an LCPA–silica composite material. The results show that aliphatic polyamines produced by diatoms for morphogenesis of their cell walls are widespread in sediments where the fossil frustules are preserved. These newly assigned components constitute useful biological markers of diatom input to sediments.

Analysis of natural diatom communities reveals novel insights into the diversity of long chain polyamine (LCPA) structures involved in silica precipitation

The intricately nanopatterned siliceous frustules of diatoms are formed under the control of template organic molecules, some of which are tightly incorporated into the frustule during formation. Long chain polyamines (LCPAs) attract dissolved silicic acid species, enhance silica precipitation and play a role in frustule nanopattern formation. LCPAs thus play a central biogeochemical role, linking the cycling of organic matter with the cycling of silicic acid in the photic zone of the upper ocean. Their distribution and molecular diversity in the environment are as yet unknown. We used high performance liquid chromatography electrospray ionization–mass spectrometry (HPLC–ESI–MSn) to investigate the distribution and molecular diversity of aliphatic LCPAs extracted from the frustules of diatoms in mixed marine plankton communities collected within the euphotic zone of the northeastern Pacific coast, Bering Sea, and Puget Sound estuary. An astonishing variety of over 100 natural LCPAS with complex distribution patterns was found in each sample. Structural variation included degree of methylation, overall chain length, presence of secondary amine functionalities, incorporation of quaternary ammonium and occurrence of sulfonated derivatives. Putrescine-based LCPAs with various degrees of methylation and N-methyl propylamine repeat units were ubiquitous. Although some LCPA species overlapped between samples, there were regional differences in composition and structural characteristics. The structural variety of LCPAs employed by diatoms to produce their nanopatterned silicified exoskeleton could inform biomimetic studies of silica precipitation.

Oligoamine analogues in combination with 2-difluoromethylornithine synergistically induce re-expression of aberrantly silenced tumour-suppressor genes

Epigenetic gene silencing is an important mechanism in the initiation and progression of cancer. Abnormal DNA CpG island hypermethylation and histone modifications are involved in aberrant silencing of tumour-suppressor genes. LSD1 (lysine-specific demethylase 1) was the first enzyme identified to specifically demethylate H3K4 (Lys4 of histone H3). Methylated H3K4 is an important mark associated with transcriptional activation. The flavin adenine dinucleotide-binding amine oxidase domain of LSD1 is homologous with two polyamine oxidases, SMO (spermine oxidase) and APAO (N1-acetylpolyamine oxidase). We have demonstrated previously that long-chain polyamine analogues, the oligoamines, are inhibitors of LSD1. In the present paper we report the synergistic effects of specific oligoamines in combination with DFMO (2-difluoromethylornithine), an inhibitor of ornithine decarboxylase, in human colorectal cancer cells. DFMO treatment depletes natural polyamines and increases the uptake of exogenous polyamines. The combination of oligoamines and DFMO results in a synergistic re-expression of aberrantly silenced tumour-suppressor genes, including SFRP2 (secreted frizzled-related protein 2), which encodes a Wnt signalling pathway antagonist and plays an anti-tumorigenic role in colorectal cancer. The treatment-induced re-expression of SFRP2 is associated with increased H3K4me2 (di-methyl H3K4) in the gene promoter. The combination of LSD1-inhibiting oligoamines and DFMO represents a novel approach to epigenetic therapy of cancer.

The Role of Proteins in Biosilicification

Although the use of silicon dioxide (silica) as a constituent of living organisms is mainly restricted to diatoms and sponges, the ways in which this process is controlled by nature continue to inspire and fascinate. Both diatoms and sponges carry out biosilificiation using an organic matrix but they adopt very different strategies. Diatoms use small and heavily modified peptides called silaffins, where the most characteristic feature is a modulation of charge by attaching long chain polyamines (LCPAs) to lysine groups. Free LCPAs can also cooperate with silaffins. Sponges use the enzyme silicatein which is homologous to the cysteine protease cathepsin. Both classes of proteins form higher-order structures which act both as structural templates and mechanistic catalysts for the polycondensation reaction. In both cases, additional proteins are continuously being discovered which modulate the process further. This paper concentrates on the role of these proteins in the biosilification process as well as in various applications, highlighting areas where focus on specific protein properties may provide further insight. The field of biosilification is a crossroads of different disciplines, where insight into the energetics and mechanisms of molecular self-assembly combine with fundamental biology, complex multicomponent colloidal systems, and an impressive array of potential technological applications.

Oriented Morphogenesis of ZnO Nanostructures from Water-Soluble Zinc Salts under Environmentally Mild Conditions and Their Optical Properties

Herein, we report a bottom-up, mineralization strategy, which borrows key principles from biomineralization processes, to synthesize nanostructured materials. A long-chain polyamine simultaneously mineralizes and assembles ZnO nanoparticles directly from water-soluble zinc salts under sustainable synthesis conditions. These thus-generated oriented structures undergo interesting morphogenesis that is controlled by changing the ratio of polyamine/Zn(2+) ions. As the ratio increases, the morphology changes from a spherical shape to oval-, dumbbell-, and finally hexagonal-rod-shaped structures that contain unique hollow rod structures. Using XPS, XRD, FT-IR, Raman spectroscopy, DLS, and confocal fluorescence microscopic analysis, we elucidate the mechanism of structural evolution; this mechanism involves the initial formation of a zinc/amine complex that is furnished with polyamine chains. These chains facilitate the condensation process to form ZnO nanoparticles and their assembly in aqueous medium at neutral pH. Further, the presence of defects in the thus-morphogenized ZnO structures leads to blue luminescence and efficient photoinduced activity, assisted by the surface-hole-trapping effect of polyamines.

Infiltration of Silica Inside Fibrillar Collagen

Diatoms frustules are created under the control of biomolecules (silaffins, silacidins and long-chain polyamines) at close to physiologic conditions.[1–4] The mechanism of biosilica formation was traditionally based on the ability of zwitterionic water-soluble proteins to create macromolecular assemblies for silica polymerization.[5–7] Recent discoveries of water-insoluble collagen matrices within certain sponge biosilica spicules[8], chitin-based scaffolds in sponge and diatom biosilica formations[9,10], as well as cingulins within diatom girdle bands[4], revive the use of insoluble biomimetic organic templates for morphogenesis of non-porous silica structures. The use of fibrillar collagen as templates for biosilica synthesis was unsuccessful in the past as only extrafibrillar silica deposition was observed.[11,12] Intrafibrillar mineralization of collagen has important implications from a biophysical perspective.[13] Here, we report a collagen biosilicification scheme based on fusion of stabilized polysilicic acid into a fluidic precursor phase upon their infiltration into polyamine-enriched collagen. The latter serves as a template and catalyst for polymerization of the precursor phase into silica that faithfully reproduces the collagen tertiary architecture. Our findings provide a new concept in biosilica materials synthesis which does not require phosphate supplements.

Isolation, Amino Acid Sequence and Biological Activities of Novel Long‐Chain Polyamine‐Associated Peptide Toxins from the Sponge Axinyssa aculeata

A novel family of functionalized peptide toxins, aculeines (ACUs), was isolated from the marine sponge Axinyssa aculeate. ACUs are polypeptides with N-terminal residues that are modified by the addition of long-chain polyamines (LCPA). Aculeines were present in the sponge extract as a complex mixture with differing polyamine chain lengths and peptide structures. ACU-A and B, which were purified in this study, share a common polypeptide chain but differ in their N-terminal residue modifications. The amino acid sequence of the polypeptide portion of ACU-A and B was deduced from 3' and 5' RACE, and supported by Edman degradation and mass spectral analysis of peptide fragments. ACU induced convulsions upon intracerebroventricular (i.c.v.) injection in mice, and disrupted neuronal membrane integrity in electrophysiological assays. ACU also lysed erythrocytes with a potency that differed between animal species. Here we describe the isolation, amino acid sequence, and biological activity of this new group of cytotoxic sponge peptides.

Molecular machines encoded by bacterially-derived multi-domain gene fusions that potentially synthesize, N-methylate and transfer long chain polyamines in diatoms

Silica glass formation in diatoms requires the biosynthesis of unusual, very long chain polyamines (LCPA) composed of iterated aminopropyl units. Diatoms processively synthesize LCPA, N-methylate the amine groups and transfer concatenated, N-dimethylated aminopropyl groups to silaffin proteins. Here I show that diatom genomes possess signal peptide-containing gene fusions of bacterially-derived polyamine biosynthetic enzymes S-adenosylmethionine decarboxylase (AdoMetDC) and an aminopropyltransferase, sometimes fused to a eukaryotic histone N-methyltransferase domain, that potentially synthesize and N-methylate LCPA. Fusions of similar, alternatively configured domains but with a catalytically dead AdoMetDC and in one case a Tudor domain, may N-dimethylate and transfer multiple aminopropyl unit polyamines onto silaffin proteins.

Biomimetic Silicification of Fibrous Chitin from Diatoms

Poly-N-acetyl-d-glucosamine (p-GlcNAc, chitin) is a biopolymer found in various organisms and in forms such as sheets, membranes, and fibers. Its presence in numerous calcium-based biominerals is well-known. Recently, chitin was even found in silica-based biominerals. While it is well-established that various proteins and even long-chain polyamines are involved in silica biomineralization events, the role of polysaccharides is rather poorly understood. Furthermore, silica-chitin nanocomposites and silica-coated chitin constructs are increasingly important, e.g., in materials science and for biomedical applications. Therefore, we have investigated the interaction between p-GlcNAc in fibers and silicic acid/silica species in vitro, using 13C and 29Si NMR as well as Raman spectroscopy, in combination with microscopic techniques. Furthermore, the kinetics of silica condensation in the presence/absence of p-GlcNAc fibers could be analyzed using the molybdenum blue test.