Lyon – INSA Team
SIX BILLION DOLLARS
SIX BILLION DOLLARS 24 000 Ferraris !
SIX BILLION DOLLARS DISINFECTANTS
WHY SO MUCH DISINFECTANT ? BIOFILM
BIOFILMS : THE INDUSTRIAL ENEMY ! More pumping power / Pressure drops Source: N. Zelver, from biofilmbook.com Leaks Maintenance + Environmental Impact A pipeline leak spews oil into a central Alberta lake A boom stretches out Source : “Microbiology control for oil and gas applications” June 2012, courtesy of Dow and Schlumberger to contain the pipeline leak. Source : Daveberta.ca Source : Calgary Herald
BIOFILMS : THE INDUSTRIAL ENEMY ! More pumping power / Pressure drops Source: N. Zelver, from biofilmbook.com Leaks Maintenance + Environmental Impact Source : “Microbiology control for oil and gas applications” June 2012, courtesy of Dow and Schlumberger Human health Source : Biomerieux
MEETING WITH INDUSTRIALISTS Cosmetics and health industry Oil industry Manufacturer of enzyme cocktails • Biofilm issues Understand • Cleaning processes • Industrial installations
EXISTING SOLUTIONS Toxic Chemical reagents Not fully Physical efficient Enzymatic Stabilization Biological cocktails
OUR OBJECTIVES - Reduce the cost - Improve the efficiency - Be Environmentally friendly
Project
FIRST STEP : DESTROY THE BIOFILM WEAPON TARGET S. aureus biofilm Biocide RBSb LYSOSTAPHIN Lyso agent LYSOSTAPHIN BioBrick™ BBa_K802000 Scattering RBSb DISPERSIN Disp Bacillus agent DISPERSIN BioBrick™ BBa_K802001
Bacillus subtilis « TORPEDO » Context Project Houry, Gohar, Deschamps, Tischenko, Aymerich, Gruss & Briandet , Proc Natl Acad Sci U S A, 2012.
Bacillus subtilis « TORPEDO » Biocide and scattering agents secreted inside the biofilm and not above, thanks to the swimming capacity of Bacillus .
CONFOCAL LASER SCANNING MICROSCOPY RESULTS CONTROL LYSOSTAPHIN S. aureus biofilm 30 µm S. aureus biofilm treated with B. subtilis + Lysostaphin BioBrick™
CONFOCAL LASER SCANNING MICROSCOPY RESULTS CONTROL DISPERSIN S. aureus biofilm S. aureus biofilm treated with B. subtilis + Dispersin BioBrick™
CONFOCAL LASER SCANNING MICROSCOPY RESULTS CONTROL LYSOSTAPHIN + DISPERSIN S. aureus biofilm BIOFILM ELIMINATION : 99 %
CONCLUSION OF THE FIRST STEP LYSOSTAPHIN + DISPERSIN : BETTER RESULT, CLEANER SURFACE KILL MODULE !
SECOND STEP Avoid new biofilm formation STICK COAT Option Option
TWO OPTIONS : WHY ? STICK Poultry and animal farming , Oil industry COAT Chemical industry
TOGGLE SWITCH SYSTEM SURFACTIN + BIOFILM BioBricks™ BBa_K802009 P xyl RBSb RBSb RBSb sfp abrB lacI
STICK Option Repression No surfactant, No biofilm repressor P xyl sfp abrB lacI IPTG STICK xylR (biofilm) P lac
COAT Option COAT Surfactant Biofilm (planktonic cells) regulator repressor P xyl sfp abrB lacI XYLOSE xylR P lac
STICK RESULTS P xyl RBSb RBSb RBSb sfp abrB lacI D abrB D abrB/ BBa_K802009 Biofilm No biofilm abrB is functional in our part
STICK : CONFOCAL MICROSCOPY RESULTS PROTECTIVE BIOFILM CONTROL E.coli biofilm • Protective B. Subtilis biofilm (red) • Colonization by E.coli (green) The protective biofilm is efficient
COAT RESULTS P xyl RBSb RBSb RBSb lacI sfp abrB Positive Supernatant Supernatant control: empty plasmid BBa_K802009 LB + SDS Surfactin production
COAT : CONFOCAL MICROSCOPY RESULTS CONTROL SURFACTIN E.coli biofilm established on Glass Slide • Surface treated with B. Subtilis supernatant + Surfactin BioBrick™. • Colonization by E. coli. The surfactin pre-treatment is efficient
CONCLUSION OF THE SECOND STEP COAT & STICK OPTIONS : Both efficient !! The surface is protected from a new biofilm formation
MODELLING : OBJECTIVES Switch control Behavior anticipation
MODELLING Physiological response Physiological response Coat Stick Stick Coat Xylose Concentration IPTG Concentration
HOW TO DEVELOP AN INDUSTRIAL SOLUTION ? Meeting with Meeting with industrialists researchers Develop an industrial process
Meeting with researchers • To improve our knowledge ...and to share this knowledge Organisation of conferences : François KEPES : SynBio pioneer in France. Romain BRIANDET : Leader of the Bioadhesion, Biofilms and Surface Hygiene lab at AgroParisTech INRA Massy (Paris).
Develop an industrial solution 1: Add BACILLUS into the tank 2: Tank cleaning ( 4 hours cycle in 6 4 1 the hydraulic system A) 3: Circuit cleaning ( 4 hours cycle FILTER 2 3 in the hydraulic system B/C) 5 4: Rinsing 5: Biological waste recovery (0.45 and 0.2 µm filter) 6: Optional: Disinfection or COAT / STICK options
Economic issues for SynBio A new economic model ? Our Reflexion • Analyze economic theories • To propose a viable economic system for iGEM and SynBio associating industrial partners
Debating with the public Outreach local community with positive applications of SynBio THE RESEARCHER’S NIGHT PUBLIC OPINION WITH A SURVEY
Outreach University members By debating with students and staff from different institutes Computer scientist Economist Master students Mechanical engineer
HUMAN PRACTICE GOALS Debating with the Economic issues for public SynBio Build a community
iGEM FRANCE First meeting initiated this year by Lyon Promote and develop : • Synbio programmation by Paris Bettencourt Governmental agencies Paris Saclay Evry • Biosafety and bioethics • SynBio teaching Lyon INSA Grenoble • Synbio Research and Bordeaux development
iGEM FRANCE First meeting initiated this year by Lyon Promote and develop : • Synbio programmation by Governmental agencies • Biosafety and bioethics • SynBio teaching • Synbio Research and development
Main Achievements A promising solution to control biofilms on closed surfaces A proposition for an Economic System for Synthetic Biology Collaboration with UBC Characterization of new parts : LYSOSTAPHIN BioBrick™ /BBa_K802000 DISPERSIN BioBrick™/BBa_K802001 SURFACTIN + BIOFILM / BBa_K802009 2 shuttle plasmids BBa_K802003 and BBa_K802004
Acknowledgments TEACHING FACILITIES INSA And all scientific, technical and administrative staff from : • Biosciences Department of INSA Lyon • MAP and BF2I laboratories • MICALIS INRA laboratory And our sponsors :
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