Novel Antibacterial Proteins from the Microbial Communities Associated with the Sponge
            <i>Cymbastela concentrica</i>
            and the Green Alga
            <i>Ulva australis</i>

Yung, Pui Yi; Burke, Catherine; Lewis, Matt; Kjelleberg, Staffan; Thomas, Torsten

doi:10.1128/aem.02038-10

Cited by 35 publications

(43 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In this respect functional metagenomic based approaches have proven useful in the identification of lipases from metagenomes from various environments, with new families being proposed [14,15]. A number of lipolytic genes have also been cloned from marine sources, with novel lipases being identified from the microbiota of the sponges Aplysina aerophoba , Hyrtios erecta and Cymbastela [16-18]. Recently fifteen different lipolytic genes, encoding proteins of between 32 and 68% amino acid identity with existing proteins in the database have been reported from a metagenomic library constructed from South China Sea marine sediment [19].…”

Section: Introductionmentioning

confidence: 99%

Isolation identification and biochemical characterization of a novel halo-tolerant lipase from the metagenome of the marine sponge Haliclona simulans

et al. 2012

View full text Add to dashboard Cite

BackgroundLipases (EC 3.1.1.3) catalyze the hydrolysis of triacyl glycerol to glycerol and are involved in the synthesis of both short chain and long chain acylglycerols. They are widely used industrially in various applications, such as baking, laundry detergents and as biocatalysts in alternative energy strategies. Marine ecosystems are known to represent a large reservoir of biodiversity with respect to industrially useful enzymes. However the vast majority of microorganisms within these ecosystems are not readily culturable. Functional metagenomic based approaches provide a solution to this problem by facilitating the identification of novel enzymes such as the halo-tolerant lipase identified in this study from a marine sponge metagenome.ResultsA metagenomic library was constructed from the marine sponge Haliclona simulans in the pCC1fos vector, containing approximately 48,000 fosmid clones. High throughput plate screening on 1% tributyrin agar resulted in the identification of 58 positive lipase clones. Following sequence analysis of the 10 most highly active fosmid clones the pCC1fos53E1 clone was found to contain a putative lipase gene lpc53E1, encoded by 387 amino acids and with a predicted molecular mass of 41.87 kDa. Sequence analysis of the predicted amino acid sequence of Lpc53E1 revealed that it is a member of the group VIII family of lipases possessing the SXTK motif, related to type C β-lactamases. Heterologous expression of lpc53E1 in E. coli and the subsequent biochemical characterization of the recombinant protein, showed an enzyme with the highest substrate specificity for long chain fatty acyl esters. Optimal activity was observed with p- nitrophenyl palmitate (C16) at 40°C, in the presence of 5 M NaCl at pH 7; while in addition the recombinant enzyme displayed activity across broad pH (3–12) and temperature (4 -60°C) ranges and high levels of stability in the presence of various solvents at NaCl concentrations as high as 5 M and at temperatures ranging from 10 to 80°C. A maximum lipase activity of 2,700 U/mg was observed with 10 mM p-nitrophenyl palmitate as substrate, in the presence of 5 mM Ca2+ and 5 M NaCl, and a reaction time of 15 min at pH 7 and 40°C; while KM and Vmax values were calculated to be 1.093 mM-1 and 50 μmol/min, respectively.ConclusionWe have isolated a novel halo tolerant lipase following a functional screen of a marine sponge fosmid metagenomic library. The activity and stability profile of the recombinant enzyme over a wide range of salinity, pH and temperature; and in the presence of organic solvent and metal ions suggests a utility for this enzyme in a variety of industrial applications.

show abstract

Section: Introductionmentioning

confidence: 99%

Isolation identification and biochemical characterization of a novel halo-tolerant lipase from the metagenome of the marine sponge Haliclona simulans

et al. 2012

View full text Add to dashboard Cite

show abstract

“…, antibacterial and antinematode) this equates to 11 active clones with unique genomic regions for the 2880 clones screened (~0.4%), which is considerably higher than previous metagenomic screens (see above). For example, a hit-rate of 0.001% was recently achieved for a screen of the metagenome created from the microbial community of U. australis [17], from which some of the isolates used in this study were derived. Thus our results would suggest that a pre-selection of bioactive-producing genomes helps with improving hit-rates and that the low abundance of organisms that encode such activities could be a limitation to the success of metagenomic screens.…”

Section: Resultsmentioning

confidence: 99%

Assessing the Effectiveness of Functional Genetic Screens for the Identification of Bioactive Metabolites

et al. 2012

Self Cite

View full text Add to dashboard Cite

A common limitation for the identification of novel activities from functional (meta) genomic screens is the low number of active clones detected relative to the number of clones screened. Here we demonstrate that constructing libraries with strains known to produce bioactives can greatly enhance the screening efficiency, by increasing the “hit-rate” and unmasking multiple activities from the same bacterial source.

show abstract

“…Microbial associates of sponges gained significance only when a remarkable similarity was found between the compounds isolated predominantly from sponges and those found in terrestrial of entirely different taxa (Perry et al 1998). The rapid development of pharmaceutical markets and technological development has increased the demand for the production of novel products from sponges toward human health concerns (Yung et al 2011). At the same time, due considerations are to given to limit and regulate the mass collection of sponges from their natural habitat as suggested by Sipkema et al (2011).…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization and bioactivity profile of the tropical sponge-associated bacterium Shewanella algae VCDB

et al. 2014

View full text Add to dashboard Cite

The pigmented, rod-shaped, Gram-negative, motile bacteria isolated from marine sponge Callyspongia diffusa exhibiting bioactivity was characterized as Shewanella algae (GenBank: KC623651). The 16S rRNA gene sequence-based phylogenetic analysis showed its similarity with the member of Shewanella and placed in a separate cluster with the recognized bacteria S. algae (PSB-05 FJ86678) with which it showed 99.0 % sequence similarity. Growth of the strain was optimum at temperature 30°C, pH 8.0 in the presence of 2.0-4.0 % of NaCl. High antibiotic activity against microbes such as Escherichia coli (MTCC 40), S. typhii (MTCC 98), P. vulgaris (MTCC 426), V. fluvialis, V. anguillarum, E. cloacae, and L. lactis was recorded. The growth of fungal pathogens such as Aspergillus niger, Aspergillus fumigatus, Saccharomyces cerevisiae, and Colletotrichum gloeosporioides was effectively controlled.

show abstract

Novel Antibacterial Proteins from the Microbial Communities Associated with the Sponge Cymbastela concentrica and the Green Alga Ulva australis

Cited by 35 publications

References 55 publications

Isolation identification and biochemical characterization of a novel halo-tolerant lipase from the metagenome of the marine sponge Haliclona simulans

Isolation identification and biochemical characterization of a novel halo-tolerant lipase from the metagenome of the marine sponge Haliclona simulans

Assessing the Effectiveness of Functional Genetic Screens for the Identification of Bioactive Metabolites

Molecular characterization and bioactivity profile of the tropical sponge-associated bacterium Shewanella algae VCDB

Contact Info

Product

Resources

About