marine ecosystems in french polynesia, a great mine for ... · lipoproteine lipase (lpl) : key...
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Marine ecosystems in French Polynesia, a great mine for
microbial metabolites of biotechnological interest
Xavier MOPPERT
Pacific Biotech Concarneau 16th October 2015
Plan
� PACIFIC BIOTECH
� Origin of strain collection
Pacific Biotech
• Exopolysaccharides
• Polyhydroxyalkanoates
• Pigments
• Secondary Metabolites with antibacterial activity
Concarneau 16th October 2015
� Conclusion and Perspectives
� Metabolites of interest
PACIFIC BIOTECH
� Created in 2006
� R&D and production activities (350k€ sales in 2014)
� A collection of over 1500 strains
Pacific Biotech Concarneau 16th October 2015
� Unique Biotechnological Start- up based on Tahiti
Strains origin
Pacific Biotech Concarneau 16th October 2015
� Kopara (microbial mats)
� Mangrove
� Deep sampling (SWAC)
Pacific Biotech
Strains origin
Concarneau 16th October 2015
� Corals and algae
� Invertebrates (Shells, Black Pearl Oysters, Urshins, …)
� Fishes , Sharks, eels, Rays…
Pacific Biotech
Strains origin
Concarneau 16th October 2015
� Production and commercialization of 3 EPS molecules (ExossineTM )
- Mo203 (Exo-HTM )
- Mo245 (Exo-TTM )
- Mo229 (Exo-PTM )
Induces hyaluronic acid production
Deep skin hydration
Pacific Biotech Concarneau 16th October 2015
Exopolysaccharides
Exopolysaccharides
Pacific Biotech
Exopolysaccharide chain
Fe2+
Nanosphere
Scanning Electronic Microscopy
EPS are good « chelators » (ion binding molecules)
Mo229 (Exo-PTM ) protects skin against pollution aggression
Concarneau 16th October 2015
Exopolysaccharides
Pacific Biotech Concarneau 16th October 2015
Cosmetic
ChylomicronsLipoproteins
Fatty acids Carried by
blood
LPL Stocked as triglycerides in adipose tissues
Lipoproteine lipase (LPL) : Key enzyme
A fall in LPL activity can also limit fat accumulation
Exopolysaccharides
Pacific Biotech Concarneau 16th October 2015
Cosmetic
High Mw RA19 EPS (3.106 Da)
Significant and dose-dependent inhibition of LPL activity
Exopolysaccharides
Pacific Biotech Concarneau 16th October 2015
Low Mw RA19 EPS (16.103 Da)
Significant and dose-dependent inhibition of LPL activity Conservation
Free-Radical depolymerization (French patent n° n°95 10045)
� A large number of scl-PHA producing bacteria
Polyhydroxyalkanoates (Biodegradable plastics)
Exploitation license
Pacific Biotech
� 3 mcl-PHA (elastomer) producing bacteria
R&D contract dealing with a special PHA for food packaging
Concarneau 16th October 2015
� Significant number of pigmented strains
Pigments
� Most interesting finding : Production of all trans - (3R, 3’R) Zeaxanthine
Pacific Biotech Concarneau 16th October 2015
Pigments
Pacific Biotech Concarneau 16th October 2015
� Macula : high concentration of Zeaxanthine
� Zeaxanthine protects eyes against UV radiation
� Only the all trans - (3R, 3’R) Zeaxanthine stereo-isomer shows anti-oxydant effect
� RA19 strain synthesizes the good stereoisomer
Advantage of Biotechnology vs chemical synthesis which would be lead to racemic mixture (yield /4)
Secondary Metabolites / antimicrobial acitivities
Pacific Biotech Concarneau 16th October 2015
Strains identified by 16 S DNA sequencing
Results
Strain Origin Sample Bacterial GenusInhibition Diameter (mm)
E. coli S. aureaus K. pneumoniae P. aeruginosa
381 Tahaa Kopara Bacillus 9 11 10 10
382 Tahaa Kopara Vibrio 10 13 -
407 Tahaa Lagoon Pseudomonas 11 - - -
550 Tahiti Eel Photobacterium - - - 10
PBT Tikehau Shell Pseudoalteromonas 10 13 11 10
747 Bora-Bora Deep Sea water(-900 m) Pseudoalteromonas - 13 9 -
804 Raivavae Kopara Pseudoalteromonas 10 11 11 -
1180 Moorea Lagoon Paracoccus 11 - - -
1200 Maiao Kopara Kosakonia - 11 - -
1249 Nuku Hiva Green Algae Bacillus 9 - 11 -
1344 Fakarava Kopara Serratia - - 12 -
1398 Fakarava Shark jaw Bacillus 11 - 9 -
� 18 % of active extracts
Antimicrobial activities
Pacific Biotech Concarneau 16th October 2015
Résultats
Souches MIC (mg/mL) vs pathogenic strainsE. coli S. aureus K. pneumoniae P. aeruginosa
382 3 ND 0,2 3407 1 ND ND ND550 ND ND ND 2PBT 0,2 0,1 1 3747 1 0,1 2 ND804 2 2 1 ND1180 0,1 ND ND ND1200 ND 1 ND ND1344 ND ND 1 ND1398 2 ND 3 ND
Pacific Biotech
Antimicrobial activities
Concarneau 16th October 2015
Spot turningup only for PBT strain
Résultats
Thin Layer Chromatography
Pacific Biotech Concarneau 16th October 2015
Strain PBT :Pseudoalteromonas piscicida sp
Results
Pacific Biotech
Activity againstPropionibacterium acne
Concarneau 16th October 2015
A few incrimated molecules :
� Depsipeptide(s)
� Glycolipide (Darapbour, et al., 2012)
� orange/brun pigment / pigmented brominated substances (Radjasa, et al., 2009)
Good candidate for more specific studies with partnership
� Modification of the growing media: Iron
Résultats PBT strain
Perspectives / Optimisation :
� Controled contamination
� Diaion HP20 resin type
Pacific Biotech Concarneau 16th October 2015
Determination of optimal growth parameters :� T° = 35°C � pH = 7,6
� Salinity = 20 g/L
µ = 0,086 h-1 G = 3,06 h
High potential of PACIFIC BIOTECH microorganism collection :
Bacterial Secondary Metabolites
AntibioticsAntifungal, …
Antifouling Bio pesticides Enzymes effectors
Conclusion and perspectives
4 strains with interesting antibacterial activity (2 Pseudoalteromonas, 1 Vibrio, 1 Paracoccus)
Pacific Biotech+ other promising metabolisms (Actinomycetes, Microalgae…)
Concarneau 16th October 2015
Pacific Biotech
Thankyou for your attention
Concarneau 16th October 2015