1. Encontro Ciência-2017, 4th July, Lisboa
INTERNATIONAL COOPERATION
Indo-Portuguese Programme for Cooperation in Science & Technology
Developing magnetic nanoparticle immobilized enzyme catalysts for biofuel applications
Suzana Ferreira-Dias & Rajeev K. Sukumaran
( )
Encontro Ciência-2017, 4th July, Lisboa

2. Tapada da Ajuda, Lisbon, Portugal
School of Agriculture, University of Lisbon
Portuguese Team:
• SUZANA FERREIRA-DIAS, PhD, 1994 (PI)
• HELENA PEREIRA, PhD , 1976 (Co PI)
• JORGE GOMINHO, PhD, 2004
• ISABEL MIRANDA, PhD, 2000
• CARLA TECELÃO, PhD, 2011
• JOANA RODRIGUES, PhD, 2016
• DUARTE NEIVA, MSc, 2009 (PhD student)
ISA
Tapada da Ajuda, Lisbon, Portugal

3. Indian Team: • RAJEEV SUKUMARAN, PhD, 2002 (PI)
• K. MADHAVAN NAMPOOTHIRI, PhD, 1997 (Co-PI)
• KV PRAJEESH, MSc, 2013 (PhD student)

4. Objectives:
Valorization of residual and waste cellulosic materials to produce biofuels and added-value compounds, using magnetic nanoparticles (MNP) immobilized enzymes, under the Biorefinery concept.

5. Valorization of residual and waste cellulosic materials:
Rice straw (India)
(c.a % cellulose, % hemicelluloses, % lignin)
Olive pomace (Portugal)
(by-product of olive oil extraction, c.a t/y in PT, with 3.3 % oil and 40 % lignocellulosic material)
De-oiled olive pomace

6. Enzyme-Catalyzed Reactions
Advantages:
Act under mild conditions
(P = 1 atm; T < 80 ̊C)
No need for co-factors
High Specificity and Selectivity
Selectivity sn-1,3 in TAG (some lipases)
Energy saving
Absence of side-reactions

7. Enzyme-Catalyzed Reactions
Constraints:
Inactivation of the biocatalyst (oxidation products; inorganic acids, phosphorus and Ni soaps; methanol, glycerine, etc.)
High price Reutilization
Immobilization in solid supports
Enzymatic processes are still more expensive than chemical processes.
Search for “low-cost” biocatalysts with high activity and operational stability.

8. Magnetic Nanoparticles (MNP) Immobilized Enzymes
Easy recovery
Easy reuse
Increased stability
Cost-effective Biomass Conversion Technology
Ren et al. BMC Biotechnology 2011, 11:63

9. MNP immobilized enzymes:
MNP- Betaglucosidases (BGL)
MNP-Cellulases
Glucose
Bioethanol
Cellulose hydrolysis
(ligno-cellulosic substrates: rice straw or de-oiled olive pomace)
Lignin
Green chemicals
Transesterification
Olive pomace oil + 2Methanol
MNP- Lipases
sn-1,3 regioselective
2 FAME Monoacylglycerols
(Biodiesel) (MAG- Emulsifiers)
(To avoid glycerol production)
Legend: FAME = Fatty Acid Methyl Esters

10. Biorefinery Concept

11. Cost-effective Biomass Conversion Technology
Specific Objectives
MNP production and Enzyme immobilization:
Production of ferro- fluid type magnetic nanoparticles (MNPs), surface modifications and characterization.
Covalent immobilization of enzymes on MNP, optimization of immobilization process, activity testing and establishment of reusability.
MNP-enzyme catalyzed reactions:
BGL performance in biomass (Rice straw or de-oiled olive pomace) hydrolysis
MNP-sn-1,3 regioselective lipases in transesterification of olive pomace oil (or rice bran oil) for biodiesel and monoglycerides (MAG; food and pharmaceutical emulsifiers).
Operational stability of MNP-immobilized enzymes.

12. Cost-effective Biomass Conversion Technology
By-products:
To look into the byproducts of biomass hydrothermal treatment, such as xylo-oligosaccharides of nutritional or pharmaceutical importance from selected Biomass (de-oiled olive pomace or rice bran/straw) and MAG production.
Scale-up of Enzymatic Processes:
Formulation of enzyme cocktails with MNP BGL conjugates and their evaluation and optimization of biomass hydrolysis (at least at 0.5 kg rice straw biomass or de-oiled olive pomace) using these enzyme blends.
Scale up of MNP-sn-1,3 regioselective lipase mediated reactions and demonstration of the transesterifcation process of ( ml) olive pomace oil with methanol for biodiesel.

13. Acknowledgements
To the Organizing Committee of Ciência-2017 for the invitation to deliver this presentation.
All of you for your time.