the microflora associated with the brown alga Ascophyllum nodosum Marjolaine Martin Microbiology and Genomics Unit Gembloux AgroBio Tech University of Liège Belgium Frankfurt Wednesday 19 ID: 205938
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Slide1
Biotechnological potential of the microflora associated with the brown alga Ascophyllum nodosum
Marjolaine Martin
Microbiology and Genomics Unit
Gembloux
Agro-Bio Tech, University of
Liège
, Belgium
Frankfurt, Wednesday 19
th
of August 2015
World Congress and Expo on Applied MicrobiologySlide2
Marine Microorganisms
Planktonic
Biofilms
Hostile environment
Molecules with
strong bioactivity
Slide3
Marine Macroalgae
Rhodophyta
Chlorophyta
Phaeophyta
Ascophyllum
nodosumSlide4
Bacteria-seaweed interactions
Anti-
microbials
Growth factors &
vitamines
Hydrolases
Review Martin
et al
, AMAB., 2014Slide5
CellulasesXylanases
Arabinanases
Amylases
Beta-
glucosidases
Agarases
Iota-
carrageenases
Kappa-
carrageenases
Estérases/Lipases
Alginate
lyases
Sulfatases
Halotolerance
Cold-active
pH-stable
Activity
Robustness
PiezotolerantSlide6
Seeking for bacterial enzymes
Functional
metagenomics
Total bacterial DNA
Culturing
Cultivable microorganismsSlide7
Ascophyllum
nodosum
Metagenomic library
from algal biofilms
Functional
Screening
Functional
MetagenomicsSlide8
Results functional metagenomicsWinter library
Summer library
180 Mb screened
100 Mb screened
> 10
carboxyesterases
1 beta-
glucosidase
1
endo
-
1,4-glucanase
6
carboxyesterases
First
metagenomic
functional screening on alga-associated bacteria
Martin
et al
, Appl.
Env
.
Microbiol
., 2014Slide9
Cold-active
Halotolerant
pH-stable
Martin
et al
, Appl.
Env
.
Microbiol
., 2014
FIRST
cellulase
identified by marine functional
metagenomics
!Slide10
functional metagenomics on alga-associated bacteria...Is a good way to identify
novel marine bacterial enzymes
Is a
great
ressource
of enzymes with biotechnological potentialIs very fastidious
when not automatedCan be problematic for the expression of some genes
Didn’t allow us the identification of
algal-polysaccharidasesSlide11
Seeking for bacterial enzymes
Functional
metagenomics
Total bacterial DNA
Culturing
Cultivable microorganismsSlide12
Screening cultivable microorganisms
324 bacteria
were isolated from
Ascophylum
nodosum
triplicatesThese 324 bacteria were screened for algal-polysaccharidases
Liquefaction
of the surrounded jellified medium
Medium is jellified with
agar, iota- or kappa-
carrageenan
, or alginate saltsSlide13
Isolated polysaccharolytic strains
Flavobacteria
(41)
Gammaproteobacteria
(37)
Flavobacteria7 Families
Algibacter, Cellulophaga, Maribacter
,
Zobellia
spp.
Cobetia
,
Colwellia
,
Marinomonas,
Paraglaciecola, Pseudoalteromonas, Shewanella, Vibrio spp.
41 acting on agars/carrageenans
23
acting on
agars/
carrageenans
26
acting
on
alginates
35
acting
on
alginates
11 strains
<97%
16S
rRNA
identities
2 strains <
97%
16S
rRNA
identities
78
polysaccharolytic strains
Novel polysaccharolytic genera
Multiple putative novel speciesSlide14
PCoA of the genera found on the three algae samples
-0.3
-0.2
-0.1
0.0
0.1
0.2
-0.2
-0.1
0.0
0.1
0.2
PCoA
- 56%
PCoA
- 44%
S
ample
1
S
ample
2
S
ample
3
Algibacter
Amphritea
Bacillus
Cellulophaga
Cobetia
Colwellia
Dokdonia
Formosa
Glaciecola
Granulosicoccus
Hydrogenophaga
Leucothrix
Loktanella
Maribacter
Marinobacter
Marinomonas
Neptunomonas
Octadecabacter
Phenylobacterium
Photobacterium
Polaribacter
Pseudoalteromonas
Pseudomonas
Psychrobacter
Psychromonas
Psychroserpens
Roseobacter
Ruegeria
Salmonella
Shewanella
Sulfitobacter
Tenacibaculum
Vibrio
Winogradskyella
Zobellia
A common core of polysaccharolytic generaSlide15
Culturing polysaccharolytic alga-associated bacteria...Shows that polysaccharolytic strains are abundant
in the
cultivable alga-associated
microflora
(25%)
Allow the identification of multiple novel polysaccharolytic speciesAllow the identification of novel polysaccharolytic genera
Suggests that cultivable polysaccharolytic strains are essential for the seaweed holobiontSlide16
Ascophyllum nodosum associated microflora...
Has been
underexplored yet
Is a great resource of
novel
enzymes with particular propertiesIs a great resource of
novel bacterial strains with biotechnological potentialSlide17
Acknowledgments
Thank YOU for your attention !
marjolaine.martin@ulg.ac.be
Gurvan Michel
Tristan
Barbeyron
Murielle Jam
Micheline Vandenbol
Promoter
Renée Martin
Michèle Nuttinck