marine ecosystems in french polynesia, a great mine for ... · lipoproteine lipase (lpl) : key...

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


� Unique Biotechnological Start- up based on Tahiti

Strains origin


� Kopara (microbial mats)

� Mangrove

� Deep sampling (SWAC)

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Strains origin


� Corals and algae

� Invertebrates (Shells, Black Pearl Oysters, Urshins, …)

� Fishes , Sharks, eels, Rays…

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Strains origin


� Production and commercialization of 3 EPS molecules (ExossineTM )

- Mo203 (Exo-HTM )

- Mo245 (Exo-TTM )

- Mo229 (Exo-PTM )

Induces hyaluronic acid production

Deep skin hydration


Exopolysaccharides

Exopolysaccharides

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Exopolysaccharide chain

Fe2+

Nanosphere

Scanning Electronic Microscopy

EPS are good « chelators » (ion binding molecules)

Mo229 (Exo-PTM ) protects skin against pollution aggression


Exopolysaccharides


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


Cosmetic

High Mw RA19 EPS (3.106 Da)

Significant and dose-dependent inhibition of LPL activity

Exopolysaccharides


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

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� 3 mcl-PHA (elastomer) producing bacteria

R&D contract dealing with a special PHA for food packaging


� Significant number of pigmented strains

Pigments

� Most interesting finding : Production of all trans - (3R, 3’R) Zeaxanthine


Pigments


� 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


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


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

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Antimicrobial activities


Spot turningup only for PBT strain

Résultats

Thin Layer Chromatography


Strain PBT :Pseudoalteromonas piscicida sp

Results

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Activity againstPropionibacterium acne


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


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…)


Pacific Biotech

Thankyou for your attention


marine ecosystems in french polynesia, a great mine for ... · lipoproteine lipase (lpl) : key...

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