Marine Sponge Axinella Research Articles

Effect of denaturing agents on a lectin isolated from marine sponge Axinella donnaniand its potential cytotoxic activity

Marine sponges are oldest metazoans which were found to have structural and biologically diverse lectins. A lectin isolated from marine sponge Axinella donnani (ADL) were evaluated for its stability against denaturing agents such as Urea, Thiourea and Guanidine hydrochloride. Urea and Thiourea decreases the stability of ADL in higher concentration (5 mM) while Guanidine hydrochloride does not show any activity. Finally, cyt otoxicity of ADL at different concentrations (0.25 500 mg/mL) was tested against MCF 7, A431 and HeLa cell lines by MTT assay. ADL exhibited potential cytotoxicity against these cell lines after 24 hours in a dose dependent manner. Thus, ADL could be a pot ent cytotoxic agent in clinical applications.

Axinellasins A-D, Immunosuppressive Cycloheptapeptide Diastereomers, Discovered via a Precursor Ion Scanning-Supercritical Fluid Chromatography Strategy from the Marine Sponge Axinella species.

The precursor ion scanning-supercritical fluid chromatography (PI-SFC) method was applied to explore new methionine sulfoxide-containing cycloheptapeptides, axinellasins A-D (1-4), from the marine sponge Axinella sp. Their structures, including absolute configurations, were elucidated by detailed spectroscopic analyses and X-ray crystallography. The total synthesis of 4 was completed via an Fmoc solid/solution-phase synthesis. Compounds 1-4 exhibited immunosuppressive effects via inhibition of T and B cell proliferation, and 1 and 4 showed better inhibitory activities than their corresponding diastereomers.

10Z‑Hymenialdisine inhibits angiogenesis by suppressing NF‑κB activation in pancreatic cancer cell lines.

10Z-Hymenialdisine is a natural product derived from the marine sponge Axinella carteri. 10Z-Hymenialdisine has anti-inflammatory effects exerted through NF-κB; however, it is unclear whether 10Z-Hymenialdisine has anti-angiogenic effects in cancer cells. In the present study, both the anti-angiogenic and antimetastatic effects of this compound in pancreatic cancer were investigated. It was initially confirmed that 10Z-Hymenialdisine significantly inhibited the proliferation of pancreatic cancer cells. Next, using both reverse transcription-quantitative PCR and ELISA, it was demonstrated that 10Z-Hymenialdisine significantly suppressed the expression of VEGF and IL-8 mRNAs and proteins in pancreatic cancer. Immunohistochemical analysis revealed that 10Z-Hymenialdisine inhibited NF-κB activity in pancreatic cancer cell lines. It was also identified that 10Z-Hymenialdisine inhibited tube formation in EA.hy926 cells. In vivo, 10Z-Hymenialdisine significantly inhibited the growth of BxPC-3 pancreatic cancer cells that were subcutaneously injected into model mice. In conclusion, the present study demonstrated that 10Z-Hymenialdisine exerted anti-angiogenic effects by suppressing NF-κB activity and angiogenic factors, such as VEGF and IL-8, in pancreatic cancer cell lines. 10Z-Hymenialdisine has potential applications as a novel therapeutic agent for the treatment of pancreatic cancer.

10Z-Hymenialdisine Inhibits Angiogenesis by Suppressing NF-κB Activation in Pancreatic Cancer Cell Lines

Introduction: 10Z-Himenialdicin is a natural product derived from the marine sponge Axinella carteri. It has reported that 10Z-Himenialdicin has an anti-angiogenetic effect through the NF-κB which produces angiogenic factors such as VEGF and IL-8. Although there is no previous report which revealed the anti-angiogenetic effects of 10Z-Himenialdicin on cancer cells, there is a potential of inhibiting the anti-angiogenetic and prevent the metastasis in pancreatic cancer.

Cultivation of fractionated cells from a bioactive-alkaloid-bearing marine sponge Axinella sp.

Sponges are among the most primitive multicellular organisms and well-known as a major source of marine natural products. Cultivation of sponge cells has long been an attractive topic due to the prominent evolutionary and cytological significance of sponges and as a potential approach to supply sponge-derived compounds. Sponge cell culture is carried out through culturing organized cell aggregates called 'primmorphs.' Most research culturing sponge cells has used unfractionated cells to develop primmorphs. In the current study, a tropical marine sponge Axinella sp., which contains the bioactive alkaloids, debromohymenialdisine (DBH), and hymenialdisine (HD), was used to obtain fractionated cells and the corresponding primmorphs. These alkaloids, DBH and HD, reportedly show pharmacological activities for treating osteoarthritis and Alzheimer's disease. Three different cell fractions were obtained, including enriched spherulous cells, large mesohyl cells, and small epithelial cells. These cell fractions were cultivated separately, forming aggregates that later developed into different kinds of primmorphs. The three kinds of primmorphs obtained were compared as regards to appearance, morphogenesis, and cellular composition. Additionally, the amount of alkaloid in the primmorphs-culture system was examined over a 30-d culturing period. During the culturing of enriched spherulous cells and developed primmorphs, the total amount of alkaloid declined notably. In addition, the speculation of alkaloid secretion and some phenomena thatoccurred during cell culturing are discussed.

Recifin A, Initial Example of the Tyr-Lock Peptide Structural Family, Is a Selective Allosteric Inhibitor of Tyrosyl-DNA Phosphodiesterase I.

Tyrosyl-DNA phosphodiesterase 1 (TDP1) is a molecular target for the sensitization of cancer cells to the FDA-approved topoisomerase inhibitors topotecan and irinotecan. High-throughput screening of natural product extract and fraction libraries for inhibitors of TDP1 activity resulted in the discovery of a new class of knotted cyclic peptides from the marine sponge Axinella sp. Bioassay-guided fractionation of the source extract resulted in the isolation of the active component which was determined to be an unprecedented 42-residue cysteine-rich peptide named recifin A. The native NMR structure revealed a novel fold comprising a four strand antiparallel β-sheet and two helical turns stabilized by a complex disulfide bond network that creates an embedded ring around one of the strands. The resulting structure, which we have termed the Tyr-lock peptide family, is stabilized by a tyrosine residue locked into three-dimensional space. Recifin A inhibited the cleavage of phosphodiester bonds by TDP1 in a FRET assay with an IC50 of 190 nM. Enzyme kinetics studies revealed that recifin A can specifically modulate the enzymatic activity of full-length TDP1 while not affecting the activity of a truncated catalytic domain of TDP1 lacking the N-terminal regulatory domain (Δ1-147), suggesting an allosteric binding site for recifin A on the regulatory domain of TDP1. Recifin A represents both the first of a unique structural class of knotted disulfide-rich peptides and defines a previously unseen mechanism of TDP1 inhibition that could be productively exploited for potential anticancer applications.

Integrating Molecular Networking and 1H NMR Spectroscopy for Isolation of Bioactive Metabolites from the Persian Gulf Sponge Axinella sinoxea.

The geographic position, highly fluctuating sea temperatures and hypersalinity make Persian Gulf an extreme environment. Although this unique environment has high biodiversity dominated by invertebrates, its potential in marine biodiscovery has largely remained untapped. Herein, we aimed at a detailed analysis of the metabolome and bioactivity profiles of the marine sponge Axinella sinoxea collected from the northeast coast of the Persian Gulf in Iran. The crude extract and its Kupchan subextracts were tested in multiple in-house bioassays, and the crude extract and its CHCl3-soluble portion showed in vitro antibacterial activity against Methicillin-resistant Staphylococcus aureus (MRSA) and Enterococcus faecium (Efm). A molecular networking (MN)-based dereplication strategy by UPLC-MS/MS revealed the presence of phospholipids and steroids, while 1H NMR spectroscopy indicated the presence of additional metabolites, such as diketopiperazines (DKPs). Integrated MN and 1H NMR analyses on both the crude and CHCl3 extracts combined with an antibacterial activity-guided isolation approach afforded eight metabolites: a new diketopiperazine, (-)-cyclo(L-trans-Hyp-L-Ile) (8); a known diketopiperazine, cyclo(L-trans-Hyp-L-Phe) (7); two known phospholipids, 1-O-hexadecyl-sn-glycero-3-phosphocholine (1) and 1-O-octadecanoyl-sn-glycero-3-phosphocholine (2); two known steroids, 3β-hydroxycholest-5-ene-7,24-dione (3) and (22E)-3β-hydroxycholesta-5,22-diene-7,24-dione (4); two known monoterpenes, loliolide (5) and 5-epi-loliolide (6). The chemical structures of the isolates were elucidated by a combination of NMR spectroscopy, HRMS and [α]D analyses. All compounds were tested against MRSA and Efm, and compound 3 showed moderate antibacterial activity against MRSA (IC50 value 70 μg/mL). This is the first study that has dealt with chemical and bioactivity profiling of A. sinoxea leading to isolation and characterization of pure sponge metabolites.

Cloning and expression of ATP N-glycosidase from the freshwater sponge Ephydatia muelleri

Previously we had discovered unusual enzymatic activity in the marine sponge Axinella polypoides, ATP N-glycosidase (Reintamm et al., 2003). We show here that the Ephydatia muelleri mRNA encoding protein with PNP_UDP_1 (phosphorylase superfamily) signature is the secreted ATP N-glycosidase. The functionality of the protein was established by recombinant expression in Pichia pastoris. In addition to the enzymatic domain, the full-length protein contains the N-terminal cysteine-rich domain belonging to the subfamily SCP_HrTT-1 (cd05559) of the SCP (sperm coating protein) superfamily (cl00133).

Two new pyrrolo-2-aminoimidazoles from a Myanmarese marine sponge, Clathria prolifera.

Marine organisms such as marine sponges and soft corals are valuable sources of pharmacologically active secondary metabolites. In our ongoing research on the discovery of new secondary metabolites from marine organisms, two new pyrrolo-2-aminoimidazoles, clathriroles A (1) and B (2), were isolated from the water-soluble portion prepared from the methanol and acetone (2:1) extract of the marine sponge, Clathria prolifera, collected in Myanmar. The chemical structures of the isolated compounds were determined using extensive spectroscopic techniques, including NMR, HRESIMS, IR, and optical rotation, and comparisons with the reported literature. The spectroscopic analyses of 1 and 2 suggested that 1 is an enantiomer of antifungal N-methylmanzacidin C isolated from the marine sponge Axinella brevistyla, whereas 2 is a diastereomer of manzacidin D at C-11 isolated from the marine sponge Astrosclera willeyana. To the best of our knowledge, this is the first report of the isolation of the pyrrolo-2-aminoimidazole compounds from C. prolifera. Furthermore, in contrast to the potency of N-methylmanzacidin C against Saccharomyces cerevisiae, the antifungal assay revealed that 1 and 2 lack any activity against this strain. Thus, these observations may suggest that the absolute configurations at both C-9 and C-11 play an important role in controlling the antifungal activity of this type of compound.

Antibacterial Activity Symbiotic Fungi of Marine Sponge Axinella sp., Aspergillus Sydowii on Four Growth Medium

Many infectious diseases caused by Escherichia coli and Staphylococcus aureus which turned into a resistant pathogen. A symbiotic fungi of marine sponge Axinella sp., Aspergillus sydowii from the waters of Riung, East Nusa Tenggara, Indonesia showed antibacterial activity, cultured on the four media, MEB (ST), Noni Juice Media (MG), avocado leaves media (AL), and Soursop leaves media (SR). The symbiotic fungi was cultured for 14 days on each media. The largest weight of symbiotic fungi biomass on ST media 138,95gr and at least 99,12gr of AL media. Purification of bioactive compound is carried out using separatory funnel, and column chromatography. The highest rendemen of extracts on SR media was 3,67%, while the lowest in ST media was 1,22%. The bioactive test used diffusion agar method. Fungi extracts from four mediums have bioactivity against, E. coli and S. aureus. The biggest inhibition zone obtained from the extract of MG KN-15-3-1-3, with inhibition zone 10.71mm and 10.98mm against E. coli and S. aureus.

Arvoredol—An unusual chlorinated and biofilm inhibiting polyketide from a marine Penicillium sp. of the Brazilian coast

Penicillium sp. F37 has been isolated from the marine sponge Axinella corrugata and shown to be closely related to Penicillium maximae. From the culture of Penicillium sp. F37 arvoredol, a novel chlorinated polyketide with 6,7-dihydro-4(5H)-benzofuranone moiety has been isolated and characterized by spectroscopic methods Arvoredol prevented biofilm formation of the human pathogen Staphylococcus epidermidis at a concentration of 125μgmL−1 by 40%. It was also active against colorectal carcinoma HCT116 cells with a MIC of 7.9μgmL−1.

Axinellin A and B: Two New Pyrrolactam Alkaloids from Axinella sp.

Two new pyrrolactam alkaloids, axinelline A (1) and B (2), were isolated from the n-BuOH extract of marine sponge Axinella sp. The structures of these compounds were elucidated on the basis of spectroscopic methods (HR-ESI-MS, 1D and 2D NMR).

Identifying a potential receptor for the antibacterial peptide of sponge Axinella donnani endosymbiont

Marine sponges and their associated bacteria are rich sources of novel secondary metabolites with therapeutic usefulness. In our earlier work, we have identified a novel antibacterial peptide from the marine sponge Axinella donnani endosymbiotic bacteria. In this work, we have carried out a comparative genomic analysis and identified a set of 60 proteins as probable receptor which is common in all the strains. The analysis on binding substrate showed that β barrel assembly machinery (BamA) of the outer membrane protein 85 (omp85) superfamily is a potential receptor protein for the antibacterial peptide. It plays a central role in OMP biogenesis, especially in cell viability. Further, the triplet and quartet motifs RGF and YGDG, respectively in L6 loop are conserved over all the strains and these conserved residues interact with antibacterial peptide to inhibit the BamA function, which is essential for OMP biogenesis.

Hexabromobenzens 농후 배양에 따른 해면(Axinella sp.) 공생 미생물의 군집구조 변화

할로겐화 또는 탈할로겐화에 관여하는 미생물을 특정하고자 해양의 주된 할로겐화합물 생산자인 해면 공생미생물의 군집구조에 미치는 HBB (hexabromobenzene)의 영향을 밝히고자 했다. 혐기 marine broth 2216에서 100ppm HBB첨가 유무에 관계없이 검출된 균주들은 산호나 해면에서 유래한 클론들과 높은 유사도를 보였으며 그 중 Deltaproteobacteria에 속하는 Desulfovibrio marinisediminis와 99% 유사도를 보이는 클론이 우점하는 것으로 나타났다. Clostridia계통의 Fusibacter paucivorans와 유사도가 높은 클론과 Lentisphaerae에 속하는 균주들은 HBB에 의해 감소하는 반면 Clostridia계통의 Vallitalea guaymasensis와 유사도가 높은 균주들은 HBB가 있는 경우에만 검출되었다. 알려진 균주들과의 비교 결과로 볼 때 D. marinisediminis 및 V. guaymasensis 계통의 균주들이 할로겐화 및 탈할로겐화에 관여하는 것으로 추정된다. Bacteria associated with marine sponges seemed to be concerned in halogenation/dehalogenation process of natural compounds. In the present study, the effect of hexabromobenzene (HBB) on the community structure of bacteria associated with a marine sponge Axinella sp. from Chuuk State under anaerobic condition was investigated. Regardless of 100 ppm HBB, most of detected microorganisms displayed high similarity with clones reported from coral or sponges. Amongst, Desulfovibrio marinisediminis like clones were dominant. Clones affiliated with Lentisphaerae and Fusibacter paucivorans (Clostridia) were detected at the conditions without HBB but clones affiliated with Vallitalea guaymasensis (Clostridia) increased its proportion with HBB. From these results and previous reports clones affiliated with D. marinisediminis and V. guaymasensis seemed to be concerned in halogenation/dehalogenation process.

Marking spherulous cell in marine sponge Axinella sp. using a 3,3′- diaminobenzidine (DAB) staining method

Debromohymenialdisine(DBH) and hymenialdisine(HD) are two bioactive alkaloids which has been isolated from marine sponge Axinella sp.living in the South China Sea,and these two alkaloids are located particularly in spherulous cells in Axinella sp..Establishment of a specific marker for this type of cells by cytochemical staining can provide an indication method for cell differentiation,and can be applied in the metabolism regulation in in vitro cell culture.The 3,3′-diaminobenzidine(DAB) staining is commonly used to detect peroxidase activity.In this study,sponge cells were fractionated and stained by DAB.A distinguishing feature of dark brown granules in cytoplasm was observed in spherulous cells.Native-PAGE was carried out with protein extracts from fresh tissue and different cell fractions of marine sponge to examine the peroxidase responsible for DAB staining.By transmission electron microscope(TEM),two kinds of inclusions were found inside the spherulous cells,the larger ones of which showed consistency in dimen-sion and amount with the positive stained granules under light microscopy.In addition,this staining method for spheru-lous cell identification was also applied in in vitro cell culture(primmorph culture).The proportion of both granularly positive-stained cells and the spherulous cells synchronously varied during a 20-day culture period,and the ratio of positive-stained spherulous cells was kept above 93%.The above results suggested that the cytochemical method of DAB staining was reliable and could be applied in both fresh-tissue dissociation and in vitro cell culture,with the dis-tinctive characteristic of positive stained granules in cytoplasm marking spherulous cells in sponge Axinella sp..

A new taurine derivative from South China Sea marine sponge Axinella sp.

A new (1) and two known (2 and 3) acylated taurine derivatives were isolated from the South China Sea marine sponge Axinella sp. The structures of the compounds were determined on the basis of spectral analysis. Compounds 1–3 did not inhibit the growth of hepatoma carcinoma cell (HepG2), lung carcinoma cell (A549), human breast carcinoma (MCF-7), no-small cell lung cancer (NCI-H460) and human nasopharyngeal carcinoma cell lines.

Donnazoles A and B from Axinella donnani sponge: very close derivatives from the postulated intermediate ‘pre-axinellamine’

Two novel dimeric pyrrole-aminoimidazole (PAI) alkaloids (−) donnazoles A–B (1–2) were isolated from the marine sponge Axinella donnani collected off the Mauritius Island. Their structures were elucidated mainly by 2D NMR. Both compounds bear the crucial substituted cyclopentane of ‘pre-axinellamine’, the hypothetical common intermediate of all dimeric PAIs. Additionally, the essential trans-6,7 ring junction of palau′amine’s congeners is already established in 1 and 2. The absolute configurations of 1 and 2 were deduced via NOE correlations in combination with the comparison of their circular dichroism data with the one of (−) sceptrin. The absolute configuration of (−) donnazoles A (1) and B (2) is coherent with the absolute configuration of the known dimeric members of PAI alkaloids.

Molecular cloning, structure, and reactivity of the second bromoperoxidase from Ascophyllum nodosum

The sequence of bromoperoxidase II from the brown alga Ascophyllum nodosum was determined from a full length cloned cDNA, obtained from a tandem mass spectrometry RT-PCR-approach. The clone encodes a protein composed of 641 amino-acids, which provides a mature 67.4kDa-bromoperoxidase II-protein (620 amino-acids). Based on 43% sequence homology with the previously characterized bromoperoxidase I from A. nodosum, a tertiary structure was modeled for the bromoperoxidase II. The structural model was refined on the basis of results from gel filtration and vanadate-binding studies, showing that the bromoperoxidase II is a hexameric metalloprotein, which binds 0.5 equivalents of vanadate as cofactor per 67.4kDa-subunit, for catalyzing oxidation of bromide by hydrogen peroxide in a bi-bi-ping-pong mechanism (kcat=153s−1, 22°C, pH 5.9). Bromide thereby is converted into a bromoelectrophile of reactivity similar to molecular bromine, based on competition kinetic data on phenol bromination and correlation analysis. Reactivity provided by the bromoperoxidase II mimics biosynthesis of methyl 4-bromopyrrole-2-carboxylate, a natural product isolated from the marine sponge Axinella tenuidigitata.

In silico biological activity of steroids from the marine sponge Axinella carteri

The steroidal composition of the marine sponge Axinella carteri, collected from Lakshadweep archipelago in the Arabian Sea was found to be a prolific source for a variety of rare steroids. The GC–MS study afforded us with the occurrence of 8 steroids belonging to four different types of nucleus, including the unique occurrence of an unconventional steroid A-norcholestan-2-one in Axinella carteri. The biological activity was predicted using the soft ware PASS. It was found that majority of the steroids extracted from Axinella carteri exhibited the potential of being agonistic to nerve growth factor (NGF).

Biological activities of ACL-I and physicochemical properties of ACL-II, lectins isolated from the marine sponge Axinella corrugata

Lectin II from the marine sponge Axinella corrugata (ACL-II) was purified by affinity chromatography on rabbit erythrocytic stroma incorporated into a polyacrylamide gel, followed by gel filtration on Ultrogel AcA 44 column. Purified ACL-II is a lectin with an Mr of 80 kDa and 78 kDa, estimated by SDS-PAGE and by FPLC on Superose 12 HR column, respectively. ACL-II mainly agglutinates native rabbit erythrocytes and this hemagglutinating activity is independent of Ca(2+), Mg(2+) and Mn(2+), but is inhibited by d-galactose, chitin and N-acetyl derivatives, with the exception of GalNAc. ACL-II is stable for up to 65 °C for 30 min, with a better stability at a pH range of 2 to 6. In contrast, ACL-I displays a strong mitogenic and cytotoxic effect.