Marine Sponges – Molecular Biology and Biotechnology References 243
Part A | 8
employing degenerate primers to target the adenylation domain of potential NRPS genes (MTF2
5
0 -GCNGGYGGYGCNTAYGTNCC-3
0 and MTR 5
0 -
CCNCGDATYTTNACYTG-3
0 [8.274]). This resulted
in the cloning of 32 putative partial NRPS genes
which, following BLAST analyses and phylogenetic
tree construction, revealed a high level of diversity with
many sharing homology (amino acid sequence identities ranging from 4099%) with genes known to
be involved in antimicrobial compound biosynthesis,
and the cloned PCR products being related to varying
degrees to gene products from at least 21 bacterial genera from 4 bacterial phyla. Examples of partial genes
cloned here, which were similar known antimicrobial
biosynthetic genes, include a sponge-derived gene sequence with similarity to the biosynthetic genes responsible for the production of the cytotoxic cyanobacterial
product, microcystin and putative NRPS genes, which
shared significant homology with genes involved in the
biosynthesis of antibacterial (fusaricidin) and antifungal (bacillomycin L, mycosubtilin) compounds.
The high levels of bacterial diversity associated
with R. ramosa together with the identification of diverse biosynthetic gene fragments of interest and the
previous isolation of bacteria showing antimicrobial activities from R. ramosa make this species of particular
research interest. Moreover, the wide diversity of potential non-ribosomal peptides that may be produced by
bacteria associated with the sponge is also especially
interesting. A PCR-based approach has recently also
been employed to demonstrate a ribosomal origin for
the bioactive metabolites polytheonamides A and B isolated from the marine sponge Theonella swinhoei. The
biosynthetic gene cluster was cloned from the sponge
metagenome, revealing a bacterial gene architecture involving six candidate enzymes which appear capable
of carrying out up to 48 posttranslational modifications.
This work, together with the work reported here, further
highlights the biosynthetic scope of ribosomal systems
within sponge metagenomes, and may ultimately provide further opportunities for peptide bioengineering
strategies [8.275].
8.7 Conclusions
The results presented here clearly identify the considerable promise of metagenomic technologies and techniques to discover and exploit novel genes and gene products with potential commercial value. However, much
work remains to be done to determine if the enzymes and
compounds discovered here can realize that potential.
References
8.1
A.C. Maloof, C.V. Rose, R. Beach, B.M. Samuels,
C.C. Calmet, D.H. Erwin, G.R. Poirier, N. Yao,
F.J. Simons: Possible animal-body fossils in preMarinoan limestones from South Australia, Nat.
Geosci. 3(9), 653–659 (2010)
8.2
E.R. Meesters, P. Knijn, R. Willemsen, G. Pennartz,
Roebers, R.W.M. van Soest: Subrubble communities
of Curaçao and Bonaire coral reefs, Coral Reefs 10,
189–197 (1991)
8.3
M.C. Diaz, B.B. Ward: Sponge-mediated nitrification in tropical benthic communities, Mar. Ecol.
Prog. Ser. 156, 97–107 (1997)
8.4
R.W. van Soest, N. Boury-Esnault, J. Vacelet,
M. Dohrmann, D. Erpenbeck, N.J. De Voogd,
N. Santodomingo, B. Vanhoorne, M. Kelly,
J.N. Hooper: Global diversity of sponges (Porifera),
PLoS ONE 7(4), e35105 (2012)
8.5
R.W.M. van Soest, N. Boury-Esnault, J.N.A. Hooper,
K. Rützler, N.J. de Voogd, B. Alvarez de Glasby,
E. Hajdu, A.B. Pisera, R. Manconi, C. Schoenberg,
D. Janussen, K.R. Tabachnick, M. Klautau, B. Picton, M. Kelly, J. Vacelet, M. Dohrmann: World
Porifera database (2012), accessed at http://www.
marinespecies.org/porifera
8.6
E. Gazave, P. Lapébie, E. Renard, J. Vacelet,
C. Rocher, A.V. Ereskovsky, D.V. Lavrov, C. Borchiellini: Molecular phylogeny restores the suprageneric subdivision of homoscleromorph sponges
(Porifera, Homoscleromorpha), PLoS ONE 12, e14290
(2010)
8.7
M.W. Taylor, R. Radax, D. Steger, M. Wagner: Sponge-associated microorganisms: Evolution, ecology, and biotechnological potential, Microbiol. Mol. Biol. Rev. 71, 295–347 (2007)
8.8
W.E.G. Müller, I.M. Müller: Origin of the metazoan
immune system: identification of the molecules
and their functions in sponges, Integr. Comp. Biol.
43, 281–292 (2003)
8.9
U. Hentschel, J. Piel, S.M. Degnan, M.W. Taylor: Genomic insights into the marine sponge
Part A | 8
employing degenerate primers to target the adenylation domain of potential NRPS genes (MTF2
5
0 -GCNGGYGGYGCNTAYGTNCC-3
0 and MTR 5
0 -
CCNCGDATYTTNACYTG-3
0 [8.274]). This resulted
in the cloning of 32 putative partial NRPS genes
which, following BLAST analyses and phylogenetic
tree construction, revealed a high level of diversity with
many sharing homology (amino acid sequence identities ranging from 4099%) with genes known to
be involved in antimicrobial compound biosynthesis,
and the cloned PCR products being related to varying
degrees to gene products from at least 21 bacterial genera from 4 bacterial phyla. Examples of partial genes
cloned here, which were similar known antimicrobial
biosynthetic genes, include a sponge-derived gene sequence with similarity to the biosynthetic genes responsible for the production of the cytotoxic cyanobacterial
product, microcystin and putative NRPS genes, which
shared significant homology with genes involved in the
biosynthesis of antibacterial (fusaricidin) and antifungal (bacillomycin L, mycosubtilin) compounds.
The high levels of bacterial diversity associated
with R. ramosa together with the identification of diverse biosynthetic gene fragments of interest and the
previous isolation of bacteria showing antimicrobial activities from R. ramosa make this species of particular
research interest. Moreover, the wide diversity of potential non-ribosomal peptides that may be produced by
bacteria associated with the sponge is also especially
interesting. A PCR-based approach has recently also
been employed to demonstrate a ribosomal origin for
the bioactive metabolites polytheonamides A and B isolated from the marine sponge Theonella swinhoei. The
biosynthetic gene cluster was cloned from the sponge
metagenome, revealing a bacterial gene architecture involving six candidate enzymes which appear capable
of carrying out up to 48 posttranslational modifications.
This work, together with the work reported here, further
highlights the biosynthetic scope of ribosomal systems
within sponge metagenomes, and may ultimately provide further opportunities for peptide bioengineering
strategies [8.275].
8.7 Conclusions
The results presented here clearly identify the considerable promise of metagenomic technologies and techniques to discover and exploit novel genes and gene products with potential commercial value. However, much
work remains to be done to determine if the enzymes and
compounds discovered here can realize that potential.
References
8.1
A.C. Maloof, C.V. Rose, R. Beach, B.M. Samuels,
C.C. Calmet, D.H. Erwin, G.R. Poirier, N. Yao,
F.J. Simons: Possible animal-body fossils in preMarinoan limestones from South Australia, Nat.
Geosci. 3(9), 653–659 (2010)
8.2
E.R. Meesters, P. Knijn, R. Willemsen, G. Pennartz,
Roebers, R.W.M. van Soest: Subrubble communities
of Curaçao and Bonaire coral reefs, Coral Reefs 10,
189–197 (1991)
8.3
M.C. Diaz, B.B. Ward: Sponge-mediated nitrification in tropical benthic communities, Mar. Ecol.
Prog. Ser. 156, 97–107 (1997)
8.4
R.W. van Soest, N. Boury-Esnault, J. Vacelet,
M. Dohrmann, D. Erpenbeck, N.J. De Voogd,
N. Santodomingo, B. Vanhoorne, M. Kelly,
J.N. Hooper: Global diversity of sponges (Porifera),
PLoS ONE 7(4), e35105 (2012)
8.5
R.W.M. van Soest, N. Boury-Esnault, J.N.A. Hooper,
K. Rützler, N.J. de Voogd, B. Alvarez de Glasby,
E. Hajdu, A.B. Pisera, R. Manconi, C. Schoenberg,
D. Janussen, K.R. Tabachnick, M. Klautau, B. Picton, M. Kelly, J. Vacelet, M. Dohrmann: World
Porifera database (2012), accessed at http://www.
marinespecies.org/porifera
8.6
E. Gazave, P. Lapébie, E. Renard, J. Vacelet,
C. Rocher, A.V. Ereskovsky, D.V. Lavrov, C. Borchiellini: Molecular phylogeny restores the suprageneric subdivision of homoscleromorph sponges
(Porifera, Homoscleromorpha), PLoS ONE 12, e14290
(2010)
8.7
M.W. Taylor, R. Radax, D. Steger, M. Wagner: Sponge-associated microorganisms: Evolution, ecology, and biotechnological potential, Microbiol. Mol. Biol. Rev. 71, 295–347 (2007)
8.8
W.E.G. Müller, I.M. Müller: Origin of the metazoan
immune system: identification of the molecules
and their functions in sponges, Integr. Comp. Biol.
43, 281–292 (2003)
8.9
U. Hentschel, J. Piel, S.M. Degnan, M.W. Taylor: Genomic insights into the marine sponge