PROJECT CODE: PN-III-P2.2.1-PED-2016-0168, Contract nr. 5PED/2017
Project type: PED (Experimental demonstration project)

Project title: Recyclable multilayer magnetic biocatalyst for synthesis of natural esters
                  Biocatalizator magnetic cu multistrat pentru sinteza in cicluri repetate a esterilor naturali
Acronym: NATMAGENZ
Project coordinator: Politehnica University of Timişoara
Project director: Prof. Dr. Eng. Francisc Peter
Budget: 600.000 RON (Public budget)
Start time: 30.01.2017
End time: 29.06.2018

PROJECT LAYOUT
The main scope of the project is to develop a demonstration model for a new biocatalyst containing a designed magnetic core and hybrid layers (organic and silica) that allow the immobilization of enzymes. The validation of the model will be accomplished through the effectiveness of the product in a specific reaction, to demonstrate that such a biocatalyst is stable, reproducible, recyclable and able to synthesize esters that are accepted as naturals according to the EC regulations (Regulation no 1334/2008 of the European Parliament and subsequent amendments).
The key objective is a comprehensive evaluation of the magnetic manipulation efficiency of enzyme functionalized magnetic nanocomposites obtained by applying cost-effective preparation procedures and manifold advanced characterization and testing techniques. The chemical composition, structure, size distribution, magneto- responsiveness and size, as well as the enzyme loading capability will be designed to fulfill the requirements for efficient biocatalysis and easy recovery of the enzyme even from viscous media, avoiding the contamination of the product and allowing its recognition as “food-grade”.   
An innovative multilayer technology will accomplish the demonstrative model. The immobilization of lipase on controlled-size magnetic core nanoparticles will be combined with stabilization of the hybrid composite through a sol-gel silica shell. The size and magnetic properties of the core particles will be adjusted to allow the optimal catalytic efficiency. This custom-made immobilized lipase will be able to catalyze the synthesis of natural esters from natural acids and natural alcohols. There is a high demand for food aroma esters recognized as naturals and the biocatalytic way is the best possibility to synthesize them. Superparamagnetic iron oxide nanoparticles (IONPs) in highly stable ferrofluid formulations will be used to fabricate functionalized magneto-responsive nanobeads for lipase immobilization, resulting in manifold reusable nanoparticle systems of high catalytic efficiency.

CONSORTIUM STRUCTURE:
Consortium leader: University Politehnica Timișoara (UPT)
Director: Prof. Dr. Eng. Francisc Peter
Members:
Dr. Eng. Anamaria Todea
Dr. Eng. Cristina Paul
Dr. Oana Marinică
Dr. Eng. Iulia Păușescu
Chem. Diana Aparaschivei (PhD student)
Chem. Corina Vasilescu (PhD student)
Biol. Horațiu Moldovan (PhD student)

Partner P1: National Institute for Research and Development of Isotopic and Molecular Technologies INCDTIM Cluj-Napoca
Responsible: Dr. Rodica Turcu
Dr. Alexander Bunge
Dr. Monica Circu
Dr. Izabella Crăciunescu
Dr. Alexandra Nan
Dr. Anca Petran
Dr. Teodora Radu

OBJECTIVES:
The objective of the project is to develop a demonstration model for a new biocatalyst containing a designed magnetic core and hybrid layers (organic and silicon) that allow the immobilization of enzymes, as well as the validation of the model through its effectiveness in a specific reaction of aroma ester synthesis.

STAGES
Stage 1 (2017, 12 month) – Development of a new multilayer magnetic biocatalyst
Stage 2 (2018, 6 month) – Synthesis of natural esters in repeated cycles using the multilayer magnetic biocatalyst


DELIVERABLES:
•    Toluene based ferrofluids with magnetite nanoparticles coated with oleic acid monolayer (mean solid size of nanoparticles approx. 7 nm; particle volume fraction: 4%, 8%, 12%);
•    High magnetization magnetic clusters functionalized with amino and carboxyl (mean size 100 nm) – grams quantities;
•    High magnetization magnetite nanoparticles (mean size 20-30 nm) functionalized with amino and carboxyl – gram quantities;
•    Size distributions for functionalized magnetic clusters and single-core magnetic nanoparticles; Laboratory scale methodology for synthesis and characterization of tailored magnetic supports for enzyme immobilization;
•    Full magnetization curves and magnetostatic properties of functionalized magnetic clusters and single-core magnetic nanoparticles;
•    Experimental methodology for preparation of the multilayer magnetic biocatalyst; Gram amounts of the model product;
•    Surface chemical composition of the functionalized magnetic nanoparticles and of the biocatalyst (atomic concentrations) and stability diagram;
•     Laboratory scale methodology for the synthesis of natural aroma esters using magnetic immobilized lipase;
•    Gram amounts of the ester products (at least three), with technical data sheet demonstrating the purity;
•    Final report, drafts of papers submitted to publication.

List of publications

Scientific Report 2017

Scientific Report 2018