Mardis de l'AIGx Philippe THONART De l'Université à l'Industrie

1. To bridge the gap between
University and Company
To bridge the gap between
University and Company
Ph. Thonart, J. Destain, M. Ongena, F. Delvigne,
J.M. Aldric, F. Weekers, S. Hiligsmann, S. Zgoulli,
P. Evrard, P. Antoine
Centre Wallon de Biologie Industrielle 1

2. Gembloux Agro Bio Tech
Passage des Déportés, 2
5030 Gembloux
Belgique
Université de Liège
Boulevard du Rectorat, 29 B40
4000 Liège
Belgique
Centre Wallon de Biologie Industrielle 2

3. CWBI - ULg
CWBI- Gembloux
Agro Biotech
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4. From cells to end products
Strain physiological
study
Strain
screening
Fermentation
technology
Metabolite or
cell harvest and
conditioning Reactors
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5. Screening on plates
Flasks
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6. 2L fermentor
20 L fermentor
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7. 500 and 2000 L fermentors
2000 L fermentor
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8. Centrifugation
Ultrafiltration
Atomisation
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9. To bridge the gap
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10. The missions
of the University
- Assistance to Community
- Teaching
- Research
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11. Precompetitive research
Transfer to the company
Applied research
Pilot production
Fundamental research
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12. The researcher must develop
a scientific strategy
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13. -Publication of research results
-Secret results
-Results protected by patents
Choices made by the researcher
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14. The University has a potential for
innovation
Innovation will require more and more
scientific knowledge
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15. * Publication of results
* Obtaining patents
Valorization of research
Popularization of results
Transfer to existing industries
integration into business strategies
Creation of spin-off
Enable innovations in non-industrialized areas
Expression of creativity by the finalization of a
product including the economic aspect
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16. Publications
To develop quality :
Impact factor, reviewed articles,
international journals, etc ..
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17. - Conferences
- Training of social environments
- Making popularization
documents
Popularization of results
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19. The CWBI develops a
new concept
Microbial ecology
applied to industry
Biodiversity
(screening new
microorganisms)
Taxonomy studies
Biopesticides
Microorganisms to
replace chemical
products
Post harvest treatment
New fermentation for
Food process
(heterofermentation)
Starters for food and
Pharmaceutical
applications
(probiotics)
New cleaning procedures
(to define the conditions
to prevent infections)
Waste water treatment with
starters (cleaning tubes,
grease trap…)
Production of
ethanol and H2
from waste water
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21. Products
Selection and production of lactic acid bacteria
- Pure cultures – Formulation - conditioning
Commercialisation with following sectors
- Pharmaceutical: food supplements (nutraceuticals)
and probiotics
- Food : starters (fermented products) and
functional foods
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22. 2222
Facilities

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24. Products
- Pure microbial cultures
- Biological activators
- Enzymes (lipase)
- Septic tank
- Starters
- Waste Water Treatment
- Plant biocontrol
- Functional microorganisms (Saccharomyces boulardii,
enrriched yeast : Se, Ca2+ ,Cu2+, Zn2+)
- Probiotics
Development of plant biocontrol : Bacillus /
Pseudozyma / Trichoderma
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25. Individual waste
water treatment plantSeptic tank
Biological
unblocker Grease traps
Products for biological
household
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26. Facilities
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Second example :
Pseudozyma
flocculosa and
Powdery Mildew Fungi
Sporothrix flocculosa; Ascomycetous
(Traquair et al., 1987)
Found in association with
clover powdery mildew
Pseudozyma flocculosa;
Basidiomycetous;
yeastlike organism
(Boekhout, et al. 2000)

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On the road to biofungicide
development
Prof. R. Belanger of the Biocontrol Laboratory at Laval
University initiated a research program on powdery
mildew biocontrol.
1970 - The tolerance of many fungal plant pathogens to
fungicide increase
1989 - Sporothrix flocculosa and regulosa were identified
as being highty effective for reducing the infection
of cucumber with powdery mildew
1995 - Sporothrix became Pseudozyma (DNA sequencing
techniques)

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Between 1989-2002
- Fundamental research and technical approach
for the Sporothrix conidies production in
collaboration with CWBI.
- Several trials were carried out under semi-
commercial conditions of greenhouse cucumber
and rose production in different countries
2000 - Patent – Methods and composition for the
biological control of plant diseases
2002-2013 : Dossier for the registration in Europe

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Netherlands
Colombia
France
Greece
…
Field trials

32. Formulation

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Action of Pseudozyma
1st hypothesis : Production of fatty acids
2nd hypothesis : Production of glycolipid - floculosin -
categorized as biosurfactants
3rd hypothesis : competition mechanism

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Conidies production in fermentors

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36. Application support and nutraceuticals
1. Vitamins for direct compression : Vitamin B1 (Thiamine), B6 (pyridoxine
hydrochloride), B12 (Cyanocobalamin) , C coated with vegetable fat
2. Microencapsulated Flavor : Microbelcaps makes a spray micro-
encapsulation of various essential oils, oleoresins, alcoholates, herbal and
natural identical molecules : lemon scent, aroma mint, apple, mango, cola,
hazelnut, yogurt, orange flavor
3. Oil rich in Omega 3 microencapsulated in acid and base resistant coatings :
microencapsulated borage oil, linseed and olive oil
4. Coated Microorganisms: Lactobacillus casei, acidophilus, bulgarius,
rhamnosus, plantarum, Bifidobacterium infantis, Streptococcus thermophilic,
Lactococcus lactis
5. Liquid papain, powder papain and coated papain
6. Dry extracts from fermented papaya : extract from fermented flavored
papaya
Products
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37. Pharmaceutical Application
1. Coated Vitamins for direct compression: Certifications: Good
Manufacturing Practice (GMP) and Drug Master File (DMF EU) : Vitamin B1,
Vitamin B6, Vitamin B12
2. Biotin 0.1%
3. Leucine
4. Vitamin C coated with vegetable fat
5. Phenylalanine
6. N-acetyl cysteine microencapsulated
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38. Facilities
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39. The company develops biological processes for the treatment and
valorisation of organic wastes, by-products of agro-food industries
and hydrocarbons.
It also realizes investment studies for the energetic valorisation of
biogas.
Biotreatment's potential is based on his skills in microbiology and
management of facilities for depollution and biomass treatment.
These are reenforced by collaborations with many academic
laboratories specialised in environmental and biochemical sciences
e.g. the Walloon Centre of Industrial Biology for microorganisms
selection, characterisation and production.
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42. Facilities
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43. Relationship between Industry-University
- Dialogue
- Respect of each partner
in economic terms,
in terms of intellectual property protection
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44. 3 axes
Microbial
biodiversity
Environment
Avenues of
collaborations
A strategy for a collaboration with
developing countries
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45. Tunisia
Senegal
Mali
Burkina Faso
Fermented milk:
M ’Bannick (Senegal)
Lben (Tunisia)
Fermented locust beans :
(Parkia biglobosa)
Netetu (Senegal)
Sumbala (Burkina Faso, Mali)
Alcohol or fruit vinegar :
Senegal, Burkina Faso,
Congo (Brazzaville)
Congo
Fermented products and
representative countries
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46. Traditional
artisan
fermentations
= natural and
spontaneous
innoculation
Empirical method
Variable duration
Variable quality
Poor hygiene
Controlled
industrial
fermentations
= add of starter
(selected
microorganisms’
culture)
Reproducible
Rapid
Quality
Organoleptic
Hygiene
Stability
Microorganisms
Raw
material
Raw
material
Fermentation
conditions
(temperature,
aeration, pH,…)
variable defined
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47. The example of Senegalese fermented milk: M’bannick
M’Bannick : a taste and a flavour that is loved by the Senegaleses
Technology: small unreliable workshops
intoxication problems
Identification of responsible germs:
Lactobacillus plantarum
Lactococcus lactis lactis
Lactococcus lactis var. diacetylactis
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48. The example of Netetu (Sénégal) –
Soumbala (Burkina Faso)
Exchange North-South
Fermented grains used as condiments
Market : 30 billions of FCFA (in Burkina)
26 thousands tons of products
Microbial biodiversity
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49. Acid flour produced by milling, fermenting and drying millet grains “pearl
millet" (Pennisetum glaucum), African cereal with small round grains,
resistant to drought.
Treatment: fermentation controlled by a lactic bacterium (Lactobacillus
plantarum), followed by drying by atomization (technique used for making
milk powder, coffee etc.)
Millet flour
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50. General diagram for production
Raw millet flour
(10% Hum, Ø 0,8 mm)

Humidification (50% water)

Adding starter mix

Fermentation during 18-20 h at 30°C

Dilution (75% water)

Atomization (230°/90°C)

Atomized acid flour
(6-8% Hum)
Bakery
mixer
Millet flour dough
Fermented flour
Manual rolling Donuts, with raisins
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51. Production of vinegar
-Development of 2 technologies
- Searching for partners
-Valorization
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52. Description of Acetobacter senegalensis sp. nov.:
 Gram-, no endospore formation
 Coccoid, 0.8 µm of width, 1.6 à 2 µm in
length and ~ 0.6 µm in Ø.
 Oxydase – and catalase +
 Optimal growth at 35°C on YGM
 Cells are opaque, convex and motile.
 G+C% of DNA is 56.0 mol %.
 Characterized by several phenotypic
features.
 The standard strain of CWBI-B418 is
LMG 23690T
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53.  By combing the phenotypic and genotypic results:
CWBI-B418 is a new species
Acetobacter senegalensis sp.nov
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54. Tank for the Orléans method
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55. - Selection of products (gum arabic, bromelain)
- Development of treatment protocols
- Transfer of raw materials towards North
- Transfer of procedures towards South
- Commercialization South-South, South-North, North-North
- Creation of spinoffs in the North and South
Avenues of collaboration
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56. Classify the gum from Burkina based on these quality
criteria (valorization South-South, South-North).
Purchase of this gum by the C.W.B.I. in order to develop
the purification procedure at industrial scale.
Economic study on the organization and the flow of gum
production in Burkina and in Wallonia.
Gum arabic
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58. Purification procedure
Bringing into solution
Filtration on filter press
Clarifying filtration on filter plates
Drying of the solution by atomization
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59. Bringing into solution
Clarifying filtration
on filter plates
Drying of the solution
by atomization
Filtration on filter press
Aggregates
Powder 59

60. Study of anthocyanes from
Hibiscus sp.
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61. Hibiscus
Malvaceae, Hibiscus sabdariffa sp
Sorrel, Roselle, sour tea, bissap
Varieties:
 Vimto: dark red calyxes
 Koor: very acidic
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62. Procedure
Extraction
Concentration
Juice containing 50% dry matter
Atomization
Instant bissap powder
I.T.A.
R.W.
I.T.A.
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63. Instant bissap bags
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65. Setting up an analytical laboratory
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66. Biological
activity
study :
analyses
(cellulose,
fatty acids,
DBO5, …
CH
4
CO
2
Biogaz
Solids
Lixiviats
-Determination of biological age
-Prediction of compaction
-Prediction of future activity
Landfill
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68. Center: sitemap of Saaba.
Periphery: different methods used for discharge collecting and composting
and landfill development
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69. Center: empty pilot landfill for the Saaba discharge.
Periphery: same landfill during filling, followed be biological activity,
compacting, laying of the mineral cover and final profile. 69

70. Innovation :
a necessity for North and South
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- From the fundamental research to a final product – a
long road
- Many biocontrol products will never satisfy the
requirement imposed on chemicals to be 95-100%
effective
- A dream or a reality. It is the problem in relation with
the fundation of spin-off company
Conclusions

72. Bronze head from Burkina Faso
(Private Collection P. Thonart)
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Art or Money