Comparative CO2 and SiO2 hydratase activity of an enzyme from the siliceous demosponge Suberitesdomuncula.

Silicase, an enzyme that catalyzes the hydrolysis of silicon-oxygen bonds, is a crucial player in breaking down silicates into silicic acid, particularly in organisms like aquatic sponges with siliceous skeletons. Despite its significance, our understanding of silicase remains limited. This study comprehensively examines silicase from the demosponge Suberites domuncula, focusing on its kinetics toward CO2 as a substrate, as well as its silicase and esterase activity. It investigates inhibition and activation profiles with a range of inhibitors and activators belonging to various classes. By comparing its esterase activity to human carbonic anhydrase II, we gain insights into its enzymatic properties. Moreover, we investigate silicase's inhibition and activation profiles, providing valuable information for potential applications. We explore the evolutionary relationship of silicase with related enzymes, revealing potential functional roles in biological systems. Additionally, we propose a biochemical mechanism through three-dimensional modeling, shedding light on its catalytic mechanisms and structural features for both silicase activity and CO2 hydration. We highlight nature's utilization of enzymatic expertise in silica metabolism. This study enhances our understanding of silicase and contributes to broader insights into ecosystem functioning and Earth's geochemical cycles, emphasizing the intricate interplay between biology and the environment.

Shallow Suberitida (Porifera, Demospongiae) from Peru.

This study describes 81 specimens belonging to Suberitida, collected during the projects Esponjas del Perú (ESPER), Esponjas da América do Sul (EsponjAS) and Semilla UCSUR 2019 (Demospongiae) along the coast of Peru, down to 30 m depth. Using morphological analyses, eight species were identified, one of which is new to science: Halichondria (H.) cristata, H. (H.) prostrata, Hymeniacidon perlevis, Johannesia reticulosa, Plicatellopsis expansa, Suberites aff. latus, Terpios cf. granulosus and Suberites inti sp. nov. Halichondria (H.) cristata, originally from Tierra del Fuego (SW Atlantic), was found widely distributed along the coast of Peru (06° S-14° S). The Magellanican H. (H.) prostrata and the formerly Chilean endemic P. expansa are extended up to Central Peru (12° S). Hymeniacidon perlevis, which presents a highly variable morphology (colour, shape, and spicule size), is firstly reported from the SE Pacific and its continuous occurrence in Peru (04° S-17° S) should be monitored given its supposed invasive potential. Johannesia reticulosa, previously known from Chile (20° S) and southern Peru (13° S), was found further north (11° S). Suberites latus and T. granulosus were originally recorded far-off from the Peruvian coast, in British Columbia and Hawaii, respectively. Thus, the occurrences of Suberites aff. latus and Terpios cf. granulosus are unexpected and should receive special attention in future molecular studies assessing their taxonomical status. Suberites inti sp. nov. characterised by its skeleton with ectosomal bouquets and multispicular choanosomal tracts, and two categories of straight tylostyles, is provisionally endemic to Paracas (13° S). With these results, the number of shallow Suberitida from Peru increases from 2 to 9. However, this number might rise as sampling in deeper environments could bring descriptions of new records.

Neosuberitenone, a New Sesterterpenoid Carbon Skeleton; New Suberitenones; and Bioactivity against Respiratory Syncytial Virus, from the Antarctic Sponge Suberites sp.

Respiratory syncytial virus (RSV) is a highly contagious human pathogen that poses a significant threat to children under the age of two, and there is a current need for new small molecule treatments. The Antarctic sponge Suberites sp. is a known source of sesterterpenes, and following an NMR-guided fractionation procedure, it was found to produce several previously unreported metabolites. Neosuberitenone (1), with a new carbon scaffold herein termed the 'neosuberitane' backbone, six suberitenone derivatives (2-7), an ansellane-type terpenoid (8), and a highly degraded sesterterpene (9), as well as previously reported suberitenones A (10) and B (11), were characterized. The structures of all of the isolated metabolites including absolute configurations are proposed on the basis of NMR, HRESIMS, optical rotation, and XRD data. The biological activities of the metabolites were evaluated in a range of infectious disease assays. Suberitenones A, B, and F (3) were found to be active against RSV, though, along with other Suberites sp. metabolites, they were inactive in bacterial and fungal screens. None of the metabolites were cytotoxic for J774 macrophages or A549 adenocarcinoma cells. The selectivity of suberitenones A, B, and F for RSV among other infectious agents is noteworthy.

A preliminary account of the Arctic/Subarctic Suberites (Porifera: Demospongiae) fauna.

Sponges of the genus Suberites are quite polymorphic and diverse, yet the delimitation of species within the group has always been challenging since there are only a few spicule types that show little, or sometimes no variation in closely allied species. Koltun (1966) created a variety, S. domuncula var. ficussomething of a dustbin assemblagewith a geographic distribution ranging from the North Atlantic, across the Arctic, to the North Pacific Oceans. Our study shows that in the Arctic/Subarctic region, boreal S. ficus is replaced by a mix of closely related species: in the western-ArcticS. lutkenii, in the eastern-ArcticS. cebriones. A defining featurecentrotylote microxeassets the northern species group apart from the boreal S. ficus and all other congeners known outside the Arctic/Subarctic seas. Altogether, we report seven species and one variety belonging to Suberites from the European Arctic/Subarctic. Five species are Arctic endemics: S. lutkenii, S. spermatozoon, S. montiniger, S. glasenappii, and S. cebriones. One species, S. virgultosus, is a typical boreal. S. syringella is apparently a species complex.

Demosponges from the sublittoral and shallow-circalittoral (lt;24m depth) Antarctic Peninsula with a description of four new species and notes on br />in situ identification characteristics.

Specimens were collected by SCUBA diving from 22 sites along the Antarctic Peninsula, spanning latitudes from Diomedea Island (62°12.185'S) to Jenny Island (67°43.325'S). The aims of the survey were to record the sponge biodiversity of the sublittoral and shallow-circalittoral and to provide information on in situ identification features. In total 24 sponge species were recorded, ranging from 0 to 12 species per site. The most widespread species were Dendrilla antarctica (10 sites), Sphaerotylus antarcticus (11 sites); and Mycale (Oxymycale) acerata (13 sites). Four species new to science were discovered and are described here: Megaciella cardenasi sp. nov., Crella (Crella) hennequinae sp. nov., Hymedesmia (Hymedesmia) noramaloneae sp. nov., Clathria (Clathria) priestleyae sp. nov. Suberites topsenti (Burton, 1929) is reassigned to Hemimycale topsenti (Burton, 1929) comb. nov. General habitat notes are provided for the survey sites, many of which were previously unsurveyed.

The Challenge of the Sponge Suberites domuncula (Olivi, 1792) in the Presence of a Symbiotic Bacterium and a Pathogen Bacterium.

Sponges, which are in close contact with numerous bacteria in prey/predator, symbiotic and pathogenic relationships, must provide an appropriate response in such situations. This starts with a discriminating recognition of the partner either by a physical contact or through secreted molecules or both. We investigated the expression of the Toll-like receptor, Caspase 3/7, Tumor Necrosis Factor receptor-associated factor 6, Bcl-2 homology protein-2 and macrophage expressed genes of axenic sponge cells in the presence of a symbiotic bacterium (Endozoicomonas sp. Hex311), a pathogen bacterium (Pseudoalteromonas sp. 1A1), their exoproducts and lipopolysaccharides. The vast majority of answers are in line with what could be observed with the symbiotic bacterium. The pathogenic bacterium seems to profit from the eukaryotic cell: suppression of the production of the antibacterial compound, inhibition of the apoptosis caspase-dependent pathway, deregulation of bacterial recognition. This work contributes new scientific knowledge in the field of immunology and apoptosis in early branching metazoan harboring within its tissue and cells a large number of symbiotic bacteria.

A new species of Acanthella Schmidt, 1862 (Porifera, Demospongiae, Dictyonellidae) from northeast region, Brazil.

Acanthella Schmidt, 1862 species are characterized by choanosomal skeleton of dendritic tracts cored by interwoven sinuously bent strongyles or strongyloxeas and echinated by straight styles, oxeas or anisoxeas (Van Soest et al. 2002). The genus Acanthella comprises 33 valid species, with six known from the Atlantic Ocean (Van Soest et al. 2018). Previously, the only record for Brazil was A. flagelliformis (Van Soest Stentoft, 1988) (Muricy 2018). Species of the genus include a transition from shrub like 'Axinellas' and herbaceous species similar to the genus Suberites Nardo, 1833 (Schmidt 1862; Vosmaer 1912). Due to its heterogeneous complex of species, Acanthella has been confused with other axinellids. Similarities were seen in genera such as Phakellia (Van Soest et al. 2002), which shares spicule complement and comparable choanosomal skeletons. In the present study, we describe a new species of Acanthella from oceanographic expeditions of the northeast region of Brazil. Two specimens of Acanthella sp. nov. were collected by dredging at Ceará and Pernambuco State, Brazil. All specimens were preserved in 92% ethanol and deposited in the Porifera Collections at the Universidade Federal de Pernambuco (UFPEPOR). The taxonomic identification was carried out through analysis of spicules morphologies, using Light Microscopy (LM) and Scanning Electron Microscopy (SEM), following methods described by Hajdu et al. (2011).

Establishment of Transgenesis in the Demosponge Suberites domuncula.

Sponges (Porifera) represent one of the most basally branching animal clades with key relevance for evolutionary studies, stem cell biology, and development. Despite a long history of sponges as experimental model systems, however, functional molecular studies are still very difficult to perform in these animals. Here, we report the establishment of transgenic technology as a basic and versatile experimental tool for sponge research. We demonstrate that slice explants of the demosponge Suberites domuncula regenerate functional sponge tissue and can be cultured for extended periods of time, providing easy experimental access under controlled conditions. We further show that an engineered expression construct driving the enhanced green fluorescence protein (egfp) gene under control of the Suberites domuncula ß-actin locus can be transfected into such tissue cultures, and that faithfully spliced transcripts are produced from such transfected DNA. Finally, by combining fluorescence-activated cell sorting (FACS) with quantitative PCR, we validate that transfected cells can be specifically reisolated from tissue based on their fluorescence. Although the number of detected enhanced green fluorescent protein (EGFP)-expressing cells is still limited, our approach represents the first successful introduction and expression of exogenous DNA in a sponge. These results represent a significant advance for the use of transgenic technology in a cornerstone phylum, for instance for the use in lineage tracing experiments.

A new species of habitat-forming Suberites (Porifera, Demospongiae, Suberitida) in the Benguela upwelling region (South Africa).

S. dandelenae sp. nov. is described from the west coast of South Africa and occurs at depths of 80-500 m among unconsolidated sediments. Specimens can reach 40 cm in length and in some areas off South Africa, up to 18 tons/km2 can be collected in a single demersal trawl. Morphologically, the sponge is straw yellow, massive with rounded lobes and has a velvety surface: it is characterized by subradiate, irregular reticulation of bundles of tylostyles and tylostrongyles. Specimens of S. dandelenae sp. nov. have three size classes of tylostyles with the largest tylostyle lengths being 516 µm (441-614 µm), medium tylostyle lengths being 352 µm (307-422 µm) and the shortest tylostyle lengths being 215 µm (153-288 µm). Apart from the presence of tylostyles, some specimens of S. dandelenae sp. nov. also possess centrotylostongyles/oxeas, tylostrongyles and microacanthostrongyles spicules. We have used morphological characters to distinguish this species and a molecular marker (cox1) to conform that all specimens are the same species. At the spicular level, S. dandelenae sp. nov. is characterized by a complex of spicule types that vary with specimen size. Following a histological investigation and re-description of the holotypes of S. ficus (Johnston, 1842) and S. tylobtusus Lévi, 1958, and comparisons with S. carnosus (Johnston, 1842), S. stilensis Burton, 1933, and other Suberites species described from the African region, it is clear that the new species is different in spicule morphology, spicule size and external morphology. For example, microacanthostrongyles are not present in S. tylobtusus and S. carnosus, whilst S. ficus possesses a second, non-spinose category of microstrongyles. Suberites tylobtusus has tylostyles that are sometimes polytylote, with heads either well formed, pear shaped or reduced, in only one size catogory. The 'tylobtuse' condition of the tylostyles is also different to the kidney-shaped and centrotylostrongyles found in S. dandelenae sp. nov. Suberites stilensis Burton, 1933 has larger and thicker tylostyles (800 µm length x 10 µm thick) than those of S. dandelenae sp. nov. A comparative analysis of partial cox1 sequences from morphologically diverse specimens of S. dandelenae sp. nov. with published material indicates that all specimens comprise a monophyletic clade. The combined morphological and genetic data support the designation of Suberites dandelenae sp. nov.

Recombinant silicateins as model biocatalysts in organosiloxane chemistry.

The family of silicatein enzymes from marine sponges (phylum Porifera) is unique in nature for catalyzing the formation of inorganic silica structures, which the organisms incorporate into their skeleton. However, the synthesis of organosiloxanes catalyzed by these enzymes has thus far remained largely unexplored. To investigate the reactivity of these enzymes in relation to this important class of compounds, their catalysis of Si-O bond hydrolysis and condensation was investigated with a range of model organosilanols and silyl ethers. The enzymes' kinetic parameters were obtained by a high-throughput colorimetric assay based on the hydrolysis of 4-nitrophenyl silyl ethers. These assays showed unambiguous catalysis with kcat/Km values on the order of 2-50 min-1 µM-1 Condensation reactions were also demonstrated by the generation of silyl ethers from their corresponding silanols and alcohols. Notably, when presented with a substrate bearing both aliphatic and aromatic hydroxy groups the enzyme preferentially silylates the latter group, in clear contrast to nonenzymatic silylations. Furthermore, the silicateins are able to catalyze transetherifications, where the silyl group from one silyl ether may be transferred to a recipient alcohol. Despite close sequence homology to the protease cathepsin L, the silicateins seem to exhibit no significant protease or esterase activity when tested against analogous substrates. Overall, these results suggest the silicateins are promising candidates for future elaboration into efficient and selective biocatalysts for organosiloxane chemistry.

Collagen from the Marine Sponges Axinella cannabina and Suberites carnosus: Isolation and Morphological, Biochemical, and Biophysical Characterization.

In search of alternative and safer sources of collagen for biomedical applications, the marine demosponges Axinella cannabina and Suberites carnosus, collected from the Aegean and the Ionian Seas, respectively, were comparatively studied for their insoluble collagen, intercellular collagen, and spongin-like collagen content. The isolated collagenous materials were morphologically, physicochemically, and biophysically characterized. Using scanning electron microscopy and transmission electron microscopy the fibrous morphology of the isolated collagens was confirmed, whereas the amino acid analysis, in conjunction with infrared spectroscopy studies, verified the characteristic for the collagen amino acid profile and its secondary structure. Furthermore, the isoelectric point and thermal behavior were determined by titration and differential scanning calorimetry, in combination with circular dichroism spectroscopic studies, respectively.

The lipopolysaccharide lipid A structure from the marine sponge-associated bacterium Pseudoalteromonas sp. 2A.

Suberites domuncula is a marine demosponge harbouring a large bacterioflora, including commensal, opportunistic and pathogenic bacteria, among which, species of the Gram-negative genus Pseudoalteromonas were identified. The sponge-bacteria interaction mechanisms are still not fully understood. As the main component of the Gram-negative bacterial outer membrane, the lipopolysaccharide (LPS) may play a role in such a crucial relationship. Moreover, the LPS is known to be the most versatile bioactive macromolecule of Gram-negative bacteria and its lipid A structure is responsible for the immunological activity of the whole LPS on eukaryotic host cells. Here it is reported the structural characterisation of the LPS lipid A moiety isolated from the S. domuncula-associated commensal bacterium, Pseudoalteromonas sp. 2A. Chemical and MALDI mass spectrometry analyses, performed on both the LPS and the isolated lipid A as well as on the intact bacterial cells, highlighted a complex family of penta-acylated lipid A species carrying two phosphate units on the disaccharide backbone.

Organic crystal lattices in the axial filament of silica spicules of Demospongiae.

The skeletal system of Demospongiae consists of siliceous spicules, which are composed of an axial channel containing an organic axial filament (AF) surrounded by a compact layer of hydrated amorphous silica. Here we report the ultrastructural investigations of the AF of siliceous spicules from two Demospongiae: Suberites domuncula and Tethya aurantium. Electron microscopy, electron diffraction and elemental mapping analyses on both longitudinal and transversal cross-sections yield that spicules's AF consist of a three-dimensional crystal lattice of six-fold symmetry. Its structure, which is the result of a biological growth process, is a crystalline assembly characterized by a lattice of organic cages (periodicity in the range of 6nm) filled with enzymatically-produced silica. In general, the six-fold lattice symmetry is reflected by the morphology of the AF, which is characterized by six-fold facets. This seems to be the result of a lattice energy minimization process similar to the situation found during the growth of inorganic crystals. Our structural exploitation of three-dimensional organic lattices generated by biological systems is expected to contribute for explaining the relation between axial filament's ultrastructure and spicule's ultimate morphology.

Tubulin Binding and Polymerization Promoting Properties of Tubulin Polymerization Promoting Proteins Are Evolutionarily Conserved.

Tubulin polymerization promoting proteins (TPPPs) constitute a eukaryotic protein family. There are three TPPP paralogs in the human genome, denoted as TPPP1-TPPP3. TPPP1 and TPPP3 are intrinsically unstructured proteins (IUPs) that bind and polymerize tubulin and stabilize microtubules, but TPPP2 does not. Vertebrate TPPPs originated from the ancient invertebrate TPPP by two-round whole-genome duplication; thus, whether the tubulin/microtubule binding function of TPPP1 and TPPP3 is a newly acquired property or was present in the invertebrate orthologs (generally one TPPP per species) has been an open question. To answer this question, we investigated a TPPP from a simple and early branching animal, the sponge Suberites domuncula. Bioinformatics, biochemical, immunochemical, spectroscopic, and electron microscopic data showed that the properties of the sponge protein correspond to those of TPPP1; namely, it is an IUP that strongly binds tubulin and induces its polymerization, proving that these features of animal TPPPs have been evolutionarily conserved.

Lipopolysaccharides from Commensal and Opportunistic Bacteria: Characterization and Response of the Immune System of the Host Sponge Suberites domuncula.

Marine sponges harbor a rich bacterioflora with which they maintain close relationships. However, the way these animals make the distinction between bacteria which are consumed to meet their metabolic needs and opportunistic and commensal bacteria which are hosted is not elucidated. Among the elements participating in this discrimination, bacterial cell wall components such as lipopolysaccharides (LPS) could play a role. In the present study, we investigated the LPS chemical structure of two bacteria associated with the sponge Suberites domuncula: a commensal Endozoicomonas sp. and an opportunistic Pseudoalteromonas sp. Electrophoretic patterns indicated different LPS structures for these bacteria. The immunomodulatory lipid A was isolated after mild acetic acid hydrolysis. The electrospray ionization ion-trap mass spectra revealed monophosphorylated molecules corresponding to tetra- and pentaacylated structures with common structural features between the two strains. Despite peculiar structural characteristics, none of these two LPS influenced the expression of the macrophage-expressed gene S. domuncula unlike the Escherichia coli ones. Further research will have to include a larger number of genes to understand how this animal can distinguish between LPS with resembling structures and discriminate between bacteria associated with it.

Functional and Structural Characterization of FAU Gene/Protein from Marine Sponge Suberites domuncula.

Finkel-Biskis-Reilly murine sarcoma virus (FBR-MuSV) ubiquitously expressed (FAU) gene is down-regulated in human prostate, breast and ovarian cancers. Moreover, its dysregulation is associated with poor prognosis in breast cancer. Sponges (Porifera) are animals without tissues which branched off first from the common ancestor of all metazoans. A large majority of genes implicated in human cancers have their homologues in the sponge genome. Our study suggests that FAU gene from the sponge Suberites domuncula reflects characteristics of the FAU gene from the metazoan ancestor, which have changed only slightly during the course of animal evolution. We found pro-apoptotic activity of sponge FAU protein. The same as its human homologue, sponge FAU increases apoptosis in human HEK293T cells. This indicates that the biological functions of FAU, usually associated with "higher" metazoans, particularly in cancer etiology, possess a biochemical background established early in metazoan evolution. The ancestor of all animals possibly possessed FAU protein with the structure and function similar to evolutionarily more recent versions of the protein, even before the appearance of true tissues and the origin of tumors and metastasis. It provides an opportunity to use pre-bilaterian animals as a simpler model for studying complex interactions in human cancerogenesis.

Potential biological role of laccase from the sponge Suberites domuncula as an antibacterial defense component.

BACKGROUND: Laccases are copper-containing enzymes that catalyze the oxidation of a wide variety of phenolic substrates. METHODS: We describe the first poriferan laccase from the marine demosponge Suberites domuncula. RESULTS: This enzyme comprises three characteristic multicopper oxidase homologous domains. Immunohistological studies revealed that the highest expression of the laccase is in the surface zone of the animals. The expression level of the laccase gene is strongly upregulated after exposure of the animals to the bacterial endotoxin lipopolysaccharide. To allow the binding of the recombinant enzyme to ferromagnetic nanoparticles, a recombinant laccase was prepared which contained in addition to the His-tag, a Glu-tag at the N-terminus of the enzyme. The recombinant laccase was enzymatically active. The apparent Michaelis constant of the enzyme is 114 µM, using syringaldazine as substrate. Exposure of E. coli to the nanoparticles, coated with Glu-tagged laccase, and to the mediator 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) in the presence of lignin, as the oxidizable substrate, resulted in an almost complete inhibition of colony formation. Quantitative studies of the effect of the laccase-coated iron oxide nanoparticles were performed using E. coli grown in suspension in reaction tubes within a magnetic nanoparticle separator. CONCLUSIONS: This newly designed magnetic nanoparticle separator allowed a removal of the nanoparticles after terminating the reaction. Using this system, a strong dose-dependent inhibition of the growth of E. coli by the laccase iron oxide nanoparticles was determined. GENERAL SIGNIFICANCE: From our data we conclude that the sponge laccase is involved in the anti-bacterial defense of the sponge organism.

Localization and characterization of ferritin in Demospongiae: a possible role on spiculogenesis.

Iron, as inorganic ion or as oxide, is widely used by biological systems in a myriad of biological functions (e.g., enzymatic, gene activation and/or regulation). In particular, marine organisms containing silica structures--diatoms and sponges--grow preferentially in the presence of iron. Using primary sponge cell culture from S. domuncula-primmorphs--as an in vitro model to study the Demospongiae spiculogenesis, we found the presence of agglomerates 50 nm in diameter exclusively inside sponge specialized cells called sclerocytes. A clear phase/material separation is observed between the agglomerates and the initial stages of intracellular spicule formation. STEM-HRTEM-EDX analysis of the agglomerates (30-100 nm) showed that they are composed of pseudohexagonal nanoparticles between 5 and 15 nm in size, displaying lattice parameters corresponding to hematite (Fe2O3) and mixed iron oxide phases typically attributed to ferritin. Further analysis, using western blotting, inductively coupled plasma mass spectrometry (ICP-MS), sequence alignment analysis, immunostaining and magnetic resonance imaging (MRI), of mature spicule filaments confirm the presence of ferritin within these organic structures. We suggest that S. domuncula can be classified as a dual biomineralizating organism, i.e., within the same cellular structure two distinct biomineralizing processes can occur as a result of the same cellular/metabolic function, spiculogenesis.

Modulation of the initial mineralization process of SaOS-2 cells by carbonic anhydrase activators and polyphosphate.

Ca-phosphate/hydroxyapatite (HA) crystals constitute the mineral matrix of vertebrate bones, while Ca-carbonate is the predominant mineral of many invertebrates, like mollusks. Recent results suggest that CaCO3 is also synthesized during early bone formation. We demonstrate that carbonic anhydrase-driven CaCO3 formation in vitro is activated by organic extracts from the demosponge Suberites domuncula as well as by quinolinic acid, one component isolated from these extracts. Further results revealed that the stimulatory effect of bicarbonate (HCO3 (-)) ions on mineralization of osteoblast-like SaOS-2 cells is strongly enhanced if the cells are exposed to inorganic polyphosphate (polyP), a linear polymer of phosphate linked by energy-rich phosphodiester bonds. The effect of polyP, administered as polyP (Ca²âº salt), on HA formation was found to be amplified by addition of the carbonic anhydrase-activating sponge extract or quinolinic acid. Our results support the assumption that CaCO3 deposits, acting as bio-seeds for Ca-carbonated phosphate formation, are formed as an intermediate during HA mineralization and that the carbonic anhydrase-mediated formation of those deposits is under a positive-negative feedback control by bone alkaline phosphatase-dependent polyP metabolism, offering new targets for therapy of bone diseases/defects.

Characterization and anti-inflammatory effects of iodinated acetylenic acids isolated from the marine sponges Suberites mammilaris and Suberites japonicus.

The previously unknown compounds 1-4, acetylenic acids with one or two iodine atom(s), were isolated from the marine sponges Suberites mammilaris and Suberites japonicus. Their complete structures were determined using NMR and mass spectrometry. The methylated compounds 1a and 2a exhibited a strong NO inhibitory effect on RAW264.7 cells, while methylated 3a and 4a were inactive in RAW264.7 cells, but highly active in BV2 microglia cells.