This operative unit focuses on the protein component contained in foodstuffs through two different methodologies of study:

• Proteomic (research line 1)
  
• Immunochimica (research line 2).

Research activity is directed toward the identification of new markers for quality, genuineness and typical characteristics, as well as toward the development of new analytical methods to ensure safety and allow for traceability of food products.

Staff: Dr.ssa Simonetta Caira, Dr.ssa Olga Fierro, Dr. Gianfranco Mamone, Dr. Gianluca Picariello

Temporary Staff: Dott.ssa Maria Grazia Calabrese (Dottorato di ricerca), Dott.ssa Annunziata Nappo (Contratto di ricerca).

Obiectives: The research activity of this group is focused on fields of nutritional research and food technology. Main aims of research are the study of safety, quality and nutritional characteristics of food productions and the set-up of new analytical methodologies. Research interests ranges from food technology to the evaluation of food quality, using new analytical techniques such as mass spectrometry coupled with chromatography and mono- and two-dimensional electrophoretic techniques and with biochemical techniques. The knowledge of food composition indicates how to improve the technological processes for their production. For this reason the effect of the technological process on food proteins, either vegetal or animal, is extensively investigated; a particular emphasis is given to proteins from milk, wheat and meat.

Research activity:

* This group is involved in the structural characterization of milk proteins and their post-translational modifications. Recently a new approach to detect phosphorylated and glycosylated caseins was developed by bidimensional electrophoresis combined with mass spectrometry. Main aim of this research is qualitative and quantitative determination of milk proteins which is directly linked to cheese yield and to the texture and flavor development in cheese.

* This group has acquired experience in the study of gluten proteins from durum and soft wheat. This research interest is focused on their technological properties and on identification of toxic peptides in food destined to celiac patients; the strategy is based on mass spectrometry techniques and immunochemical tests.

* Molecular protein markers of the meat proteolysis during ripening have been individuated by a proteomic approach.

* A research line is focused on new methodologies for the synthesis and chemical characterization of molecules with biological activity and of analogous of bioactive peptides for their possible pharmacological applications. Moreover, solid phase synthesis technologies are used for the preparation of antigenic peptides utilizable for production of antibodies in host animals

* This group investigates on the proteolytic activity of lactic bacteria during ripening of dairy products and their role in casein degradation, giving rise to bioactive peptides. The specific effect of proteases from cell-wall proteins is studied by proteomic analysis.

Selected Pubblications

A. Cereda, F. Forlani, S. Iametti, R. Bernhardt, P. Ferranti, G. Picariello, S. Pagani, F. Bonomi. Molecular recognition between Azotobacter vinelandii rhodanese and a sulfur acceptor protein.

Biol. Chem., in press

Mamone G, Caira S, Garro G, Nicolai A, Ferranti P, Picariello G, Malorni A, Chianese L, Addeo F.Casein phosphoproteome: identification of phosphoproteins by combined mass spectrometry and two-dimensional gel electrophoresis.

Electrophoresis. 2003 Aug;24(16):2824-37.

Caira S, Ferranti P, Gatti M, Fornasari ME, Barone F, Lilla S, Mucchetti G, Picariello G, Chianese L, Neviani E, Addeo F. Synthetic peptides as substrate for assaying the proteolytic activity of Lactobacillus helveticus.

J Dairy Res. 2003 Aug;70(3):315-25.

Dettin M, Ferranti P, Scarinci C, Picariello G, Di Bello C. Is the V3 loop involved in HIV binding to CD4?

Biochemistry. 2003 Aug 5;42(30):9007-12

Lilla S, Caira S, Ferranti P, Addeo F. Mass spectrometric characterisation of proteins in rennet and in chymosin-based milk-clotting preparations.

Rapid Commun Mass Spectrom. 2001;15(13):1101-12.

Basile A, Ferranti P, Pocsfalvi G, Mamone G, Miraglia N, Caira S, Ambrosi L, Soleo L, Sannolo N, Malorni A. A novel approach for identification and measurement of hemoglobin adducts with 1,2,3,4-diepoxybutane by liquid chromatography/electrospray ionisation mass spectrometry and matrix-assisted laser desorption/ionisation tandem mass spectrometry.

Rapid Commun Mass Spectrom. 2001;15(8):527-40.

Metafora V, Franco P, Massa O, Morelli F, Stiuso P, Ferranti P, Mamone G, Malorni A, Stoppelli MP, Metafora S. Phosphorylation of seminal vesicle protein IV on Ser58 enhances its peroxidase-stimulating activity.

Eur J Biochem. 2001 Jul;268(13):3858-69.

Ferranti P, Mamone G, Malorni A. Preparation and mass spectrometric analysis of S-nitrosohemoglobin.

Methods Mol Biol. 2000;146:147-65

Mamone G, Ferranti P, Chianese L, Scafuri L, Addeo F. Qualitative and quantitative analysis of wheat gluten proteins by liquid chromatography and electrospray mass spectrometry. Rapid Commun Mass Spectrom. 2000;14(10):897-904.

Evidente A, Sparapano L, Fierro O, Bruno G, Motta A. Sapinofuranones A and B, two new 2(3H)-dihydrofuranones produced by sphaeropsis sapinea, a common pathogen of conifers. J. Nat. Prod. 1999 Feb; 62(2):253-6.

Staff: Dr. Rosa Pizzano

Temporary Staff: Dr. Carla Manzo (Dottorato di Ricerca)

Food matrices are extremely complex. They range from liquids and pastes to solids and powers, whose components may have been processed by a variety of heat treatments or procedures of extraction. In addition, different chemical modifications may occur even in the finished product depending on packaging and storage conditions. Therefore, application of immunochemical techniques to most of the specific aspects of food analysis requires tailor-made antibodies for use as reagents. Our analytical approach is based on the development of antipeptide antibodies at predetermined specificity toward native or chemically modified protein substructures, properly designed for each set purpose. This strategy proved to be successful in resolving a number of specific problems concerning milk and cheese analysis. By combining immunochemical techniques using antipeptide antibodies with conventional biochemical methods we were able to propose new analytical procedures in order to:

a) characterize the protein component of a genuine product and distinguish it from the adulterated ones (1,3,5);

b) identify protein markers of dairy products related to the technology employed (2,4,6,7);

c) recognize the protein components involved in allergies to milk and related products (results in course of publication);

d) follow the release of bioactive peptides by enzymatic proteolysis during cheese ripening (results in course of publication).

Pubblications Cited:

1. PIZZANO R., NICOLAI M.A., ADDEO F.

Antipeptide antibodies as analytical tools to discriminate among bovine alfa-s1-casein components

J. Agric. Food Chem. 46, 766-771 (1998)

2. PIZZANO R., NICOLAI M.A., SICILIANO R., ADDEO F.

Specific detection of the Amadori compounds in milk by using polyclonal antibodies raised against a lactosylated peptide

J. Agric. Food Chem. 46, 5373-5379 (1998)

3. PIZZANO R., NICOLAI M.A., ADDEO F.

Antipeptide antibodies in milk and cheese analysis

in "Recent Research Developments in Agricultural and Food Chemistry", Pandalai S. G. ed., Research Signpost, Trivandrum, India, Vol. 3, 403-412 (1999)

4. PIZZANO R., NICOLAI M.A., PADOVANO P., FERRANTI P., BARONE F., ADDEO F.

Immunochemical evaluation of bovine beta-casein and its 1-28 phosphopeptide in cheese during ripening

J. Agric. Food Chem. 48, 4555-4560 (2000)

5. FERRANTI P., PIZZANO R., GARRO G., CAIRA S., CHIANESE L., ADDEO F.

Mass spectrometry-based procedure for the identification of ovine casein heterogeneity

J. Dairy Res. 68, 35-51 (2001)

6. PIZZANO R., NICOLAI M.A., CHIANESE L., ADDEO F.

Detection of plasmin-mediated beta-casein hydrolysis in milk and cheese through immunochemical techniques

in "Recent Research Developments in Agricultural and Food Chemistry", Pandalai S. G. ed., Research Signpost, Trivandrum, India, Vol. 5, 191-199 (2001)

7. PIZZANO R., NICOLAI M.A., SICILIANO R., MANZO C., ADDEO F.

Immunochemical Detection of Formylated gamma 2-Casein in Cheese

J. Agric. Food Chem. (submitted, 2003)

On going projects:

Actually the UO is involved in two financed projects both supported by the Ministry of University, Scientific Research and Technology .

GEFORPASTA: This project is aimed to detection of molecular markers formed during heat and/or chemical treatments in foods for individuation of frauds in milk and dairy field.

SIFORTI: This project is aimed to development of a methodology for qualitative and quantitative detection of bacterial toxins in milk and dairy products by immunochemical techniques and mass spectrometry.

http://www.pnragrobio.unina.it

Collaborations:

Dipartimento di Scienza degli Alimenti - Università "Federico II", Portici (NAPOLI)

Dipartimento Di Scienze Molecolari Agroalimentari, Università di Milano

Dipartimento di Processi Chimici per l'Ingegneria, Università degli Studi di Padova

Menarini Ricerche S.p.A. Pomezia (Roma)

Amylum Group, Aalst, Belgium