Italian Forum on Industrial Biotechnology and Bioeconomy IFIB 2014 Genoa, 25-26 September 2014 #IFIB14 THE PROTEIN FACTORY Elena Rosini, Giovanni Bernardini, Flavia Marinelli, Loredano Pollegioni, Gabriele Candiani, Paola D’Arrigo, Stefano Servi, Sergio Riva The Protein Factory Research Center - Università degli studi dell’Insubria, Politecnico di Milano, Istituto di Chimica del Riconoscimento Molecolare CNR – via Mancinelli 7 20131 Milano, Italy - phone: +390223993125 - +390223993062 e-mail: [email protected] Presentation – “The Protein Factory” Research Center has been established at the Politecnico of Milano (Dept. CMIC G. Natta), the University of Insubria in Varese (Dept. Biotechnology and Life Sciences) and the CNR in Milan (Institute of Chemistry of Molecular Recognition) to promote basic and applied research activities on proteins of biotechnological relevance. The Protein Factory (comprising 16 research units) reaches the critical size to manage complex projects and to offer specific services in a strategic sector of the industrial and biomedical research. Detailed information can be found at the website: http://www.dbsm.uninsubria.it/proteinf/ Abstract – The activities are coordinated by the “core lab” located at the Dept. CMIG G. Natta of Politecnico di Milano, via Mancinelli 7: this lab identifies, develops and produces the proteins for the different biotechnological applications (1). Proteins and processes developed at “The Protein Factory”: 1. For the pharmaceutical field: setup of bioconversion processes, e.g. for the direct conversion of cephalosporin C into 7-amino cephalosporanic acid and the modification of complex antibiotic molecules; fermentative production of antimicrobial glycopeptides, lipodepsipeptides, lantibiotics and strain improvement of the producing strains; new antiinfective drugs designed on the basis of emerging resistance mechanisms in the producer microbes; new drugs for the treatment of schizophrenia. 2. For the chemical field: novel, engineered enzymes with altered substrate specificity (1); multi-step enzyme catalyzed synthesis of chiral natural glycerol and amino acid derivatives; dynamic kinetic resolution of amino acids and amino acid esters; asymmetric synthesis of amino-group-containing complex molecules; isolation and production of phospholipase D for phosphatidyl serine production; NP-enzymes. 3. For the diagnostic/analytical field: production of biosensors for the analysis of the neuromodulator D-serine in brain, to detect amino acids, amines, herbicides in foods and biological samples; production of a stable cholesterol oxidase useful in clinical chemistry. 4. For the environmental field: evolution of an enzyme to degrade/quantify the herbicide glyphosate; biorefineries: identification and production of new chitinases, peroxidases and laccases (by metagenomics and protein engineering approaches) for recalcitrant biomass treatment. 5. For the biomedical field: production of an “ad hoc” biocatalyst for an enzyme pro-drug cancer therapy and synthesis of magnetic nanoparticles combined with such an enzyme (2); production of human proteins related to important pathologies: -synuclein, Hsp70, DAAO, pLG72, PRODH; antigenic proteins for vaccine production (3). 6. System biocatalysis: artificial in vitro metabolism (4). References 1. Pollegioni, L.; Molla, G. Trends Biotechnol. 2011, 29, 276-283. 2. Bava, A.; Gornati, R.; Cappellini, F.; Caldinelli, L.; Pollegioni, L.; Bernardini, G. Nanomedicine 2013, 8, 1797-1806. 3. Piubelli, L.; Campa, M.; Temporini, C.; Binda, E.; Mangione, F.; Amicosante, M.; Terreni, M.; Marinelli, F.; Pollegioni, L. Microb Cell Fact. 2013, 12, 115. 4. http://www.cost.eu/domains_actions/cmst/Actions/CM1303 Elena Rosini The Protein Factory Research Center - Università degli studi dell’Insubria, Politecnico di Milano, Istituto di Chimica del Riconoscimento Molecolare CNR – via Mancinelli 7, 20131 Milano, Italy - phone: +390223993125/3062 e-mail: [email protected]; [email protected] “The Protein Factory” Research Center (Director: prof. Loredano Pollegioni) has been established at the Politecnico of Milano (Dept. CMIC G. Natta), the University of Insubria in Varese (Dept. Biotechnology and Life Sciences) and the CNR in Milan (Institute of Chemistry of Molecular Recognition) to promote basic and applied research activities on proteins of biotechnological relevance. The Protein Factory Research Center has reached the critical size for managing complex projects and offering tailored services to meet emerging needs in industrial, environmental and biomedical research. Detailed information can be accessed at the website: http://www.theproteinfactory.it • • • • • • • • Animal Biotechnology and Aquaculture Applied Organic Chemistry Biocatalysis BioCell Biomaterials Cell Biology Chemical Biotechnologies Chemistry and Characterization of Innovative Polymers • Computer Simulations and Spectroscopy • Gene Delivery • Membrane Proteins • Molecular Genetics • Protein Engineering • Protein Production • Stereoselective Synthesis • Structural Biology NMR The Research Center The aim of “The Protein Factory” consists in offering an integrated laboratory for the production and characterization of native and engineered recombinant proteins useful in the different fields of biotechnology. The “core lab” is mainly focused on protein engineering, protein production, fermentation, chemical biology, and cell biology. The specific expertises and skills of the 16 research units allow to contribute to the progress of translational research within industry and medicine. The activities are coordinated by the “core lab” (Dept. CMIG G. Natta of Politecnico di Milano, via Mancinelli 7, Milano) Proteins produced/engineered at “The Protein Factory” • Acylases (glutaryl acylase, cephalosporin acylase, lipoglycopeptide and lipodepsipeptide acylases) • Oxidases (D-amino acid oxidase, L-amino acid oxidase, glycine oxidase, proline oxidase, cholesterol oxidase, histamine oxidase, diamine oxidase) • Proteases (bacterial proteases, HIV-1 protease, D,D-carboxypeptidases) • Cellulases, chitinases, peroxidases (manganese and versatile), laccases, accessory lignin degrading enzymes • Aminotransferases, phospholipase D • Mycobacterium tuberculosis antigens (Ag85B, TB10.4) • Human proteins: pLG72, α-synuclein, Hsp70, D-amino acid oxidase, Phox2B