Identification of a suitable sterilisation method for collagen derived from a marine Demosponge

Collagen is widely used as a biomedical material, and its importance is likely to grow as research and understanding progresses in this field. As a biomedical material, ensuring the sterility of collagen before use as, or incorporation into, a medical device is paramount. However, common sterilisation techniques can induce changes in the physical structure and protein chemistry of collagen, potentially affecting the performance. In this preliminary study, the influence of autoclaving, gamma irradiation and ethylene oxide gas sterilisation on the denaturation temperature and helical content of the collagen was evaluated using differential scanning calorimetry and Fourier transform infrared spectroscopy. Early results indicate that all sterilisation techniques affect collagen properties but suggest that the least damaging of the techniques investigated was y irradiation.

Environmental selection on transcriptome-derived SNPs in a high gene flow marine fish, the Atlantic herring (Clupea harengus)

High gene flow is considered the norm for most marine organisms and is expected to limit their ability to adapt to local environments. Few studies have directly compared the patterns of differentiation at neutral and selected gene loci in marine organisms. We analysed a transcriptome-derived panel of 281 SNPs in Atlantic herring (Clupea harengus), a highly migratory small pelagic fish, for elucidating neutral and selected genetic variation among populations and to identify candidate genes for environmental adaptation. We analysed 607 individuals from 18 spawning locations in the northeast Atlantic, including two temperature clines (5-12 °C) and two salinity clines (5-35‰). By combining genome scan and landscape genetic analyses, four genetically distinct groups of herring were identified: Baltic Sea, Baltic-North Sea transition area, North Sea/British Isles and North Atlantic; notably, samples exhibited divergent clustering patterns for neutral and selected loci. We found statistically strong evidence for divergent selection at 16 outlier loci on a global scale, and significant correlations with temperature and salinity at nine loci. On regional scales, we identified two outlier loci with parallel patterns across temperature clines and five loci associated with temperature in the North Sea/North Atlantic. Likewise, we found seven replicated outliers, of which five were significantly associated with low salinity across both salinity clines. Our results reveal a complex pattern of varying spatial genetic variation among outlier loci, likely reflecting adaptations to local environments. In addition to disclosing the fine scale of local adaptation in a highly vagile species, our data emphasize the need to preserve functionally important biodiversity.

Alkaline protease production by marine fungus engyodontium BTMFS 10

The thesis entitled “Alkaline Protease Production by Marine Fungus Engyodontium BTMFS 10”.Proteases are the single class of enzymes, which occupy a pivotal position with respect to their application in both physiological and commercial filed. Protease in the industrial market is expected to increase further in the coming year. The current trend is to use microbial enzymes since they provide a greater diversity of catalytic activities and can be produced more economically. Main objective of theses studies are the optimization of various physicochemical factors in the solid state fermentation for the production of alkaline protease enzyme, characterization of the enzyme, evaluation of the enzyme for various industrial application. The result obtained the during the course of theses study indicate the scope for the utilization of this study Marine Fungus E. Album for extra cellular protease production employing solid state fermentation

Production, purification, genetic characterization and application studies of tannase enzyme from marine fungus Aspergillus awamori

Marine fungi remain totally unexplored as a source of industrial enzyme and prospective applications. Further tannase production by a marine organism has so far not been established. The primary objective of this study included the evaluation of the potential of Aspergillus awamori isolated from sea water as part of an earlier study and available in the culture collection of the Microbial technology laboratory for tannase production through different fermentation methods, optimization of bioprocess variables by statistical methods, purification and characterization of the enzyme, genetic study, and assessment of its potential applications.

Lipase production by marine fungus Aspergillus Awamori Nagazawa

The present study indicate the scope for the utilization of the marine fungus Aspergillus awamori Nagazawa BTMFW 032 for extracellular lipase production employing submerged fermentation. To the best of our knowledge this is the first report on lipase production by a marine fungus employing statistical modeling towards industrial production. The characterization of purified lipase produced by A. awamori showed stability in organic solvents, oxidizing agent and reducing agents, I,3-regiospecificity and hydrolytic activity. These properties make this lipase an ideal candidate for biocatalysis in organic media for the production of novel compounds such as biodiesel and sugar fatty esters. 91.4 % reduction in oil and grease content in ayurvedic oil by the treatment of A. awamori lipase indicates that there is a scope for this enzyme in the treatment of oil effluents and bioremediation. There is ample scope for further research on the biochemistry of the enzyme, structure elucidation and enzyme engineering towards a wide range of further applications, besides enriching scientific knowledge on marine enzymes.

Molecular cloning of alkaline protease gene from marine fungus Engyodontium album

Department of Biotechnology, Cochin University of Science and Technology

Molecular cloning and homology modelling of a subtilisin-like serine protease from the marine fungus, Engyodontium album BTMFS10

An alkaline protease gene (Eap) was isolated for the first time from a marine fungus, Engyodontium album. Eap consists of an open reading frame of 1,161 bp encoding a prepropeptide consisting of 387 amino acids with a calculated molecular mass of 40.923 kDa. Homology comparison of the deduced amino acid sequence of Eap with other known proteins indicated that Eap encode an extracellular protease that belongs to the subtilase family of serine protease (Family S8). A comparative homology model of the Engyodontium album protease (EAP) was developed using the crystal structure of proteinase K. The model revealed that EAP has broad substrate specificity similar to Proteinase K with preference for bulky hydrophobic residues at P1 and P4. Also, EAP is suggested to have two disulfide bonds and more than two Ca2? binding sites in its 3D structure; both of which are assumed to contribute to the thermostable nature of the protein.

Utilization of prawn waste for chitinase production by the marine fungus Beauveria bassiana by solid state fermentation

Prawn waste, a chitinous solid waste of the shell®sh processing industry, was used as a substrate for chitinase production by the marine fungus Beauveria bassiana BTMF S10, in a solid state fermentation (SSF) culture. The process parameters in¯uencing SSF were optimized. A maximum chitinase yield of 248.0 units/g initial dry substrate (U/gIDS) was obtained in a medium containing a 5:1 ratio (w/v) of prawn waste/sea water, 1% (w/w) NaCl, 2.5% (w/w) KH2PO4, 425±600 lm substrate particle size at 27 °C, initial pH 9.5, and after 5 days of incubation. The presence of yeast extract reduced chitinase yield. The results indicate scope for the utilization of shell®sh processing (prawn) waste for the industrial production of chitinase by using solid state fermentation.

Molecular cloning and homology modelling of a subtilisin-like serine protease from the marine fungus, Engyodontium album BTMFS10

An alkaline protease gene (Eap) was isolated for the first time from a marine fungus, Engyodontium album. Eap consists of an open reading frame of 1,161 bp encoding a prepropeptide consisting of 387 amino acids with a calculated molecular mass of 40.923 kDa. Homology comparison of the deduced amino acid sequence of Eap with other known proteins indicated that Eap encode an extracellular protease that belongs to the subtilase family of serine protease (Family S8). A comparative homology model of the Engyodontium album protease (EAP) was developed using the crystal structure of proteinase K. The model revealed that EAP has broad substrate specificity similar to Proteinase K with preference for bulky hydrophobic residues at P1 and P4. Also, EAP is suggested to have two disulfide bonds and more than two Ca2? binding sites in its 3D structure; both of which are assumed to contribute to the thermostable nature of the protein.

Vegetation and environmental dynamics in the southern Black Sea region since 18 kyr BP derived from the marine core 22-GC3

Sediments from the Black Sea, a region historically dominated by forests and steppe landscapes, are a valuable source of detailed information on the changes in regional terrestrial and aquatic environments at decadal to millennial scales. Here we present multi-proxy environmental records (pollen, dinoflagellate cysts, Ca, Ti and oxygen isotope data) from the uppermost 305 cm of the core 22-GC3 (42°13.53′N, 36°29.55′E) collected from a water depth of 838 m in the southern part of the Black Sea in 2007. The records span the last ~ 18 kyr (all ages are given in cal kyr BP). The pollen data reveal the dominance of the Artemisia-steppe in the region, suggesting rather dry/cold environments ~ 18–14.5 kyr BP. Warming/humidity increase during melt-water pulses (~ 16.1–14.5 kyr BP), indicated by δ18O records from the 22-GC3 core sediment and from the Sofular Cave stalagmite, is expressed in more negative δ13C values from the Sofular Cave, usually interpreted as the spreading of C3 plants. The records representing the interstadial complex (~ 14.5–12.9 kyr BP) show an increase in temperature and moisture, indicated by forest development, increased primary productivity and reduced surface run-off, whereas the switch from primary terrigenous to primary authigenic Ca origin occurs ~ 500 yr later. The Younger Dryas cooling is clearly demonstrated by more negative δ13C values from the Sofular Cave and a reduction of pines. The early Holocene (11.7–8.5 kyr BP) interval reveals relatively dry conditions compared to the mostly moist and warm middle Holocene (8.5–5 kyr BP), which is characterized by the establishment of the species-rich warm mixed and temperate deciduous forests in the low elevation belt, temperate deciduous beech-hornbeam forests in the middle and cool conifer forest in upper mountain belt. The border between the early and middle Holocene in the vegetation records coincides with the opening of the Mediterranean corridor at ~ 8.3 kyr BP, as indicated by a marked change in the dinocyst assemblages and in the sediment lithology. Changes in the pollen assemblages indicate a reduction in forest cover after ~ 5 kyr BP, which was likely caused by increased anthropogenic pressure on the regional vegetation.

Integrated marine gravity field in the Brazilian coast from altimeter-derived sea surface gradient and shipborne gravity

Sea surface gradients derived from the Geosat and ERS-1 satellite altimetry geodetic missions were integrated with marine gravity data from the National Geophysical Data Center and Brazilian national surveys. Using the least squares collocation method, models of free-air gravity anomaly and geoid height were calculated for the coast of Brazil with a resolution of 2` x 2`. The integration of satellite and shipborne data showed better statistical results in regions near the coast than using satellite data only, suggesting an improvement when compared to the state-of-the-art global gravity models. Furthermore, these results were obtained with considerably less input information than was used by those reference models. The least squares collocation presented a very low content of high-frequency noise in the predicted gravity anomalies. This may be considered essential to improve the high resolution representation of the gravity field in regions of ocean-continent transition. (C) 2010 Elsevier Ltd. All rights reserved.

Biotransformation of alpha-Bromoacetophenones by the Marine Fungus Aspergillus sydowii

The biotransformation reactions of alpha-bromoacetophenone (1), p-bromo-alpha-bromoacetophenone (2), and p-nitro-alpha-bromoacetophenone (3) by whole cells of the marine fungus Aspergillus sydowii Ce19 have been investigated. Fungal cells that had been grown in artificial sea water medium containing a high concentration of chloride ions (1.20 M) catalysed the biotransformation of 1 to 2-bromo-1-phenylethanol 4 (56%), together with the alpha-chlorohydrin 7 (9%), 1-phenylethan-1,2-diol 9 (26%), acetophenone 10 (4%) and phenylethanol 11 (5%) identified by GC-MS analysis. In addition, it was observed that the enzymatic reaction was accompanied by the spontaneous debromination of 1 to yield alpha-chloroacetophenone 5 (9%) and alpha-hydroxyacetophenone 6 (18%) identified by GC-FID analysis. When 2 and 3 were employed as substrates, various biotransformation products were detected but the formation of halohydrins was not observed. It is concluded that marine fungus A. sydowii Ce19 presents potential for the biotransformations of bromoacetophenone derivatives.

Filamentous fungi from the Atlantic marine sponge Dragmacidon reticulatum

Dragmacidon reticulatum is a marine sponge of wide occurrence in the Eastern and Western Atlantic. Little is known about D. reticulatum fungal diversity. Filamentous fungi recovered from D. reticulatum were assessed in the present study using a polyphasic taxonomic approach, including classical morphology, molecular biology and MALDI-TOF ICMS. Ninety-eight fungal strains were isolated from two D. reticulatum samples by using six different culture media, which were identified up to the genus level. Sixty-four distinct fungal ribotypes were obtained, distributed among twenty-four different genera belonging to the Ascomycota and Zygomycota. Representatives of Penicillium and Trichoderma were the most diverse and abundant fungi isolated. Amongst Penicillium spp. three isolates belonged to the same ribotype can be considered as a putative new species. Data derived from the present study highlight the importance of using a polyphasic approach to get an accurate identification in order to structure a reliable culture collection. © 2012 Springer-Verlag Berlin Heidelberg.