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 Industrielle1

Gembloux Agro Bio TechPassage des Déportés, 2

5030 GemblouxBelgique

Université de LiègeBoulevard du Rectorat, 29 B40

4000 LiègeBelgique

Centre Wallon de Biologie Industrielle2

CWBI - ULg

CWBI- Gembloux Agro Biotech

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From cells to end products

Strain physiological study

Strain

screening

Fermentation technology

Metabolite or cell harvest and

conditioning Reactors

. ... .

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Screening on plates

Flasks

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2L fermentor

20 L fermentor

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500 and 2000 L fermentors

2000 L fermentor

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Centrifugation

Ultrafiltration

Atomisation

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To bridge the gap

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The missions of the University

- Assistance to Community

- Teaching

- Research

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Precompetitive research

Transfer to the company

Applied research

Pilot production

Fundamental research

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The researcher must develop

a scientific strategy

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-Publication of research results

-Secret results

-Results protected by patents

Choices made by the researcher

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The University has a potential for innovation

Innovation will require more and more

scientific knowledge

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* Publication of results* Obtaining patents

Valorization of research

Popularization of results

Transfer to existing industriesintegration 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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Publications

To develop quality :

Impact factor, reviewed articles, international journals, etc ..

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- Conferences

- Training of social environments

- Making popularizationdocuments

Popularization of results

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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 forFood 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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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) andfunctional foods

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Facilities

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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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Individual waste water treatment plantSeptic tank

Biological unblocker Grease traps

Products for biologicalhousehold

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Facilities

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Second example : Pseudozyma

flocculosa and Powdery Mildew Fungi

Sporothrix flocculosa; Ascomycetous(Traquair et al., 1987)

Found in association withclover powdery mildew

Pseudozyma flocculosa; Basidiomycetous; yeastlike organism

(Boekhout, et al. 2000)

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On the road to biofungicidedevelopment

Prof. R. Belanger of the Biocontrol Laboratory at LavalUniversity initiated a research program on powderymildew biocontrol.

1970 - The tolerance of many fungal plant pathogens tofungicide increase

1989 - Sporothrix flocculosa and regulosa were identifiedas being highty effective for reducing the infectionof cucumber with powdery mildew

1995 - Sporothrix became Pseudozyma (DNA sequencingtechniques)

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Between 1989-2002 - Fundamental research and technical approachfor the Sporothrix conidies production incollaboration with CWBI.

- Several trials were carried out under semi-commercial conditions of greenhouse cucumberand rose production in different countries

2000 - Patent – Methods and composition for thebiological control of plant diseases

2002-2013 : Dossier for the registration in Europe

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Netherlands

Colombia

France

Greece

…

Field trials

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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Application support and nutraceuticals

1. Vitamins for direct compression : Vitamin B1 (Thiamine), B6 (pyridoxinehydrochloride), B12 (Cyanocobalamin) , C coated with vegetable fat

2. Microencapsulated Flavor : Microbelcaps makes a spray micro-encapsulation of various essential oils, oleoresins, alcoholates, herbal andnatural 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 flavoredpapaya

Products

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Pharmaceutical Application

1. Coated Vitamins for direct compression: Certifications: GoodManufacturing 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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Facilities

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The company develops biological processes for the treatment andvalorisation of organic wastes, by-products of agro-food industriesand hydrocarbons.

It also realizes investment studies for the energetic valorisation ofbiogas.

Biotreatment's potential is based on his skills in microbiology andmanagement of facilities for depollution and biomass treatment.These are reenforced by collaborations with many academiclaboratories specialised in environmental and biochemical sciencese.g. the Walloon Centre of Industrial Biology for microorganismsselection, characterisation and production.

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Facilities

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Relationship between Industry-University

- Dialogue

- Respect of each partner

in economic terms,

in terms of intellectual property protection

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3 axes

Microbial biodiversity

Environment

Avenues of collaborations

A strategy for a collaboration with developing countries

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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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Traditional artisan fermentations

= natural and spontaneous innoculation

Empirical methodVariable durationVariable qualityPoor hygiene

Controlled industrial fermentations

= add of starter (selected

microorganisms’ culture)

ReproducibleRapid

Quality Organoleptic

HygieneStability

Microorganisms

Raw material

Raw material

Fermentationconditions

(temperature,aeration, pH,…)

variable defined

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The example of Senegalese fermented milk: M’bannick

M’Bannick : a taste and a flavour that is loved by the Senegaleses

Technology: small unreliable workshopsintoxication problems

Identification of responsible germs:

Lactobacillus plantarum

Lactococcus lactis lactis

Lactococcus lactis var. diacetylactis

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The example of Netetu (Sénégal) –Soumbala (Burkina Faso)

Exchange North-SouthFermented grains used as condiments

Market : 30 billions of FCFA (in Burkina)26 thousands tons of products

Microbial biodiversity

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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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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)

Bakerymixer

Millet flour dough

Fermented flour

Manual rolling Donuts, with raisins50

Production of vinegar

-Development of 2 technologies

- Searching for partners

-Valorization

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Description of Acetobacter senegalensis sp. nov.:

Gram-, no endospore formation

Coccoid, 0.8 µm of width, 1.6 à 2 µm inlength 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 phenotypicfeatures.

The standard strain of CWBI-B418 isLMG 23690T

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By combing the phenotypic and genotypic results:

CWBI-B418 is a new species

Acetobacter senegalensis sp.nov

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Tank for the Orléans method

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- 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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Classify the gum from Burkina based on these qualitycriteria (valorization South-South, South-North).

Purchase of this gum by the C.W.B.I. in order to developthe purification procedure at industrial scale.

Economic study on the organization and the flow of gumproduction in Burkina and in Wallonia.

Gum arabic

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Purification procedure

Bringing into solution

Filtration on filter press

Clarifying filtration on filter plates

Drying of the solution by atomization

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Bringing into solution

Clarifying filtration

on filter plates

Drying of the solution

by atomization

Filtration on filter pressAggregates

Powder59

Study of anthocyanes from Hibiscus sp.

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Hibiscus

Malvaceae, Hibiscus sabdariffa sp

Sorrel, Roselle, sour tea, bissap

Varieties:

Vimto: dark red calyxes

Koor: very acidic

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Procedure

Extraction

Concentration

Juice containing 50% dry matter

Atomization

Instant bissap powder

I.T.A.

R.W. I.T.A.

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Instant bissap bags

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Setting up an analytical laboratory

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Biological activity study : analyses(cellulose, fatty acids,

DBO5, …

CH4

CO2

Biogaz

Solids

Lixiviats-Determination of biological age

-Prediction of compaction

-Prediction of future activity

Landfill

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Center: sitemap of Saaba. Periphery: different methods used for discharge collecting and composting and landfill development

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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

Innovation :

a necessity for North and South

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- From the fundamental research to a final product – along road

- Many biocontrol products will never satisfy therequirement imposed on chemicals to be 95-100%effective

- A dream or a reality. It is the problem in relation withthe fundation of spin-off company

Conclusions

Bronze head from Burkina Faso(Private Collection P. Thonart)

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Art or Money