Dep. of Soil, Plant and Food Sciences

The effect of some quality parameters of the milk (refrigeration
time, pH, protein, and fat/protein ratio) on
the extent of acidification in the production technology
of table Mozzarella without starters was investigated.
A screening phase carried out at the laboratory level
demonstrated that variations of the milk characteristics
require different levels of acidification to keep constant
the quality of the cheese. The elaboration of the data
collected throughout the successive experimentation
on industrial scale allowed us to find a mathematical
model to describe the relationships between the pH of
the curd at stretching time and the milk characteristics,
of which the protein concentration and the refrigeration
time play the main roles.

The assessment of the origin and quality of raw materials is pivotal for the protection and valorization of typical dairy products. Italian high moisture mozzarella cheese, also called " Fiordilatte " , is manufactured by stretching in hot water the curd, which can be produced inside the dairy (fresh curd) or purchased as semi-finished product from specialized companies (pre-made stored curd). The employment of stored curd allows cheese manufacturers to obtain economic profit thanks to the reduction of the production costs. The use of the semi-finished product has not to be mentioned in the label according to the actual EU regulation, and this has given rise to competition between local milk farmers and curd-producing companies causing misinformation among the consumers, and unfair competition to the disadvantage of the traditional dairies. In the present research, a proteomic approach was used to investigate " Fior-dilatte " produced with or without the employment of stored curd. The results obtained allowed us to identify a molecular marker which is present in very small amounts in cheese made from fresh curd and at high levels in cheese made from stored curd. The marker is a s1-I casein (fragment 24e199), the main product of primary proteolysis formed by the action of chymosin on a s1 casein. It can be easily detected by electrophoresis in the presence of urea (urea-PAGE) and is measured by image analysis. Even though the kinetic of formation of this proteolytic product has been well known for several decades, in mozzarella it has a peculiar behavior compared to other cheeses. As a matter of fact, it originates during the first part of cheesemaking, and has a very slow increase during storage of the cheese, due to the denaturation of the clotting enzyme by the hot water stretching process. The results obtained demonstrated that the quantification of a s1-I fragment should be recommended to guarantee the production of " Fiordilatte " obtained from fresh milk under normal conditions, and could be the basis for the development of a method able to fully protect the traditional cheesemaking procedure.

An experimental investigation was carried out with the aim of appraising the effects of natural tocopherols on lipid oxidation in extruded breakfast cereals. Corn flakes produced on an industrial scale, with and without added tocopherols and stored in commercial packing under shelf conditions, were compared using sensory evaluation and analyses of volatile compounds and triacylglycerol polymerization products. The sensory evaluation showed the effectiveness of tocopherols in limiting the development of off-flavour during storage. The analysis of the volatile compounds, focused on the secondary products of the lipid oxidation, showed that the antioxidant activity of tocopherols limited the evolution of the headspace towards a more intense oxidation, notably the further oxidation of hexanal into hexanoic acid (that reached 32% and 15% in the flakes, respectively, without and with added tocopherols, after 360 days). The percentage of hexanoic acid in the headspace positively correlated with almost all the descriptors of the defect considered for the sensory evaluation. The analysis of the triacylglycerol polymerization products confirmed the inhibiting action of tocopherols on lipid oxidative degradation.

Preliminarily, 146 strains of yeasts were screened for the antifungal activity toward the indicator Penicillium
roqueforti DPPMAF1. The strain Meyerozyma guilliermondii LCF1353 was selected and used for
dough fermentation. The water/salt soluble extract of the dough was analyzed by HPLC and GC/MSSPME.
The synthesis of the extracellular cell wall-degrading enzyme b-1,3-glucanase and ethyl-acetate
was shown. The effect on conidia germination mainly suggested a fungistatic activity. M. guilliermondii
LCF1353 was used as starter for dough fermentation in combination with Wickeramomyces anomalus
1695 and Lactobacillus plantarum 1A7, which were previously selected for antifungal activity. The growth
of the strains was monitored by plate count and molecular techniques, and competitive or antagonistic
interactions among them were excluded. Bread started with the combination of M. guilliermondii
LCF1353,W. anomalus LCF1695 and L. plantarum 1A7 showed a more prolonged shelf life compared to the
other breads. Fungal growth was delayed at least until 14 days of storage, under conditions of high
artificial inoculum. The bread manufactured with the above combination showed good chemical and
textural characteristics and, as shown by sensory analysis, it was appreciated for elasticity, color and
taste.

Capillary electrophoresis was used to characterize glutenin proteins from ancient varieties of Southern Italy common wheat and to determine the technological properties of wheat flours based on a glutenin index. Three zones were identified in the electropherograms, indicated as A, B, and C according to electroelution order. The three zones corresponded to the low molecular weight glutenin subunits and to the y-and x-type high molecular weight subunits, respectively. The ratio B/C was correlated to the alveographic parameter P/L. These results indicated that flours resulting in a B/C ratio lower than 2 produced elastic doughs whereas flours resulting in a B/C ratio higher than 2 produced doughs more resistant to extension. This study showed that capillary electrophoresis is useful for determining the types and quantities of gluten proteins in the evaluation of wheat-flour technological properties of a limited number of noncommercial varieties as evidenced by the x-type content which seems to strongly influence the flour technological parameters.

Ciauscolo is a short-ripened fermented sausage manufactured in the Marche region (central Italy) that has recently received a protected geographical indication product classification (PGI). The aim of this study was the exploration of the biochemical traits of this traditional Italian salami, with a special focus on protein and lipid composition. Ciauscolo salami was characterized by pH of 5.1 and 0.91 water activity. A prevalence of lactic acid bacteria in the microbiota was found. The free amino acids and biogenic amines average content was 2657 and 255 mg/kg, respectively. With regards to lipids composition unsaturated fatty acids represented 63% and 72% of total and free fatty acids. Despite these results had wide statistical variability, attributable to differences in the processing parameters and raw matter used, some peculiar traits were found: (1) structural muscular proteins underwent to less proteolysis than sarcoplasmic ones; (2) glycogen phosphorylase, enolase, and aldolase were the most proteolyzed among the sacoplasmic proteins; (3) there was inverse correlation between histamine content and yeasts population, and a direct correlation between the gly-pro content and lactic acid bacteria counts; (4) the content of aspartic acid and methyonine seem to be a possible molecular marker able to distinguish between double and single milling.

The NDUFS4 subunit of complex I of the mammalian respiratory chain has a fully conserved carboxy-terminus with a canonical RVSTK phosphorylation site. Immunochemical analysis with specific antibodies shows that the serine in this site of the protein is natively present in complex I in both the phosphory-lated and non-phosphorylated state. Two-dimensional IEF/SDS–PAGE electrophoresis, 32 P labelling and immunodetection show that ''in vitro " PKA phosphorylates the serine in the C-terminus of the NDUFS4 subunit in isolated bovine complex I. 32 P labelling and TLC phosphoaminoacid mapping show that PKA phosphorylates serine and threonine residues in the purified heterologous human NDUFS4 protein.

The assessment of the origin and quality of raw materials is pivotal for the protection and valorization of typical dairy products. Italian high moisture mozzarella cheese, also called " Fiordilatte " , is manufactured by stretching in hot water the curd, which can be produced inside the dairy (fresh curd) or purchased as semi-finished product from specialized companies (pre-made stored curd). The employment of stored curd allows cheese manufacturers to obtain economic profit thanks to the reduction of the production costs. The use of the semi-finished product has not to be mentioned in the label according to the actual EU regulation, and this has given rise to competition between local milk farmers and curd-producing companies causing misinformation among the consumers, and unfair competition to the disadvantage of the traditional dairies. In the present research, a proteomic approach was used to investigate " Fior-dilatte " produced with or without the employment of stored curd. The results obtained allowed us to identify a molecular marker which is present in very small amounts in cheese made from fresh curd and at high levels in cheese made from stored curd. The marker is a s1-I casein (fragment 24e199), the main product of primary proteolysis formed by the action of chymosin on a s1 casein. It can be easily detected by electrophoresis in the presence of urea (urea-PAGE) and is measured by image analysis. Even though the kinetic of formation of this proteolytic product has been well known for several decades, in mozzarella it has a peculiar behavior compared to other cheeses. As a matter of fact, it originates during the first part of cheesemaking, and has a very slow increase during storage of the cheese, due to the denaturation of the clotting enzyme by the hot water stretching process. The results obtained demonstrated that the quantification of a s1-I fragment should be recommended to guarantee the production of " Fiordilatte " obtained from fresh milk under normal conditions, and could be the basis for the development of a method able to fully protect the traditional cheesemaking procedure.

In this study, a starter culture was tentatively
made by testing autochthonous Lactobacillus and Staphylococcus
strains, isolated from traditional ‘Salsiccia Sarda’
sausage. Three sausage-making trials were carried out using
an association of Staphylococcus xylosus ? Lactobacillus
curvatus (A), Staphylococcus xylosus ? Lactobacillus
plantarum (B), and Lactobacillus plantarum alone (C).
Microbiological profiles, free amino acids, biogenic amines,
and free fatty acids were evaluated after 21 days of ripening.
Sausages A and B showed the highest number of lactic
acid bacteria that produced a rapid acidification of substrates
and reduced the number of spoilage microorganisms.
Sausages made with L. plantarum alone or together with
S. xylosus had the highest content of free amino acids,
whereas the one produced with S. xylosus ? L. curvatus
showed the lowest content of total biogenic amines and best
acceptance score. No significant differences among samples
were found as to the free fatty acids concentration.

The effects of probiotic supplementation on hen egg quality were studied, focusing on yolk protein changes.
Electrophoretic analysis and protein spot identification were performed and the relative quantity of each protein was
computed. Results showed a higher quantity of ovotransferrin, YGP42 and YGP40 (P<0.05) together with a reduced
quantity of lipovitellin-2 (P<0.01) in the yolk of treated hens. Noteworthy is the increased concentration of YGP40.
This is a yolk glycoprotein of 40 kDa recently suggested to be a potential new allergen. Eggs from treated hens also
had a lower fat content than those from the control hens, both because of a lower fat concentration (P<0.05) and a
smaller yolk (P<0.01).

This article appeared in a journal published by Elsevier. The attached copy is furnished to the author for internal non-commercial research and education use, including for instruction at the authors institution and sharing with colleagues. Other uses, including reproduction and distribution, or selling or licensing copies, or posting to personal, institutional or third party websites are prohibited. In most cases authors are permitted to post their version of the article (e.g. in Word or Tex form) to their personal website or institutional repository. Authors requiring further information regarding Elsevier's archiving and manuscript policies are encouraged to visit: http://www.elsevier.com/copyright a b s t r a c t Autochthonous lactic acid bacteria from emmer flour were screened based on the kinetic of acidification and used to ferment beverages containing emmer flour, emmer gelatinized flour, and emmer malt at percentages ranging 5e30% (wt/wt). Preliminarily, the concentration of raw flour and malt was selected based on sensory analysis. Different protocols were set up for the manufacture of four different beverages which used Lactobacillus plantarum 6E as the starter. Emmer beverages were mainly differentiated based on the concentration of organic acids, carbohydrates, amino acids, dietary fibers, vitamins, antioxidant and phytase activities, and volatiles and sensory profiles. Wheat flour bread was used as the control to determine the hydrolysis index (HI ¼ 100), as an indirect estimation of the glycemic index. The beverage made with 30% (wt/wt) of gelatinized flour showed an HI of 56%, its viscosity was improved by using an EPS-producing strain and it allowed the survival of the potential probiotic Lactobacillus rhamnosus SP1 at cell density of ca. 5 Â 10 8 cfu/ml throughout storage at 4 C. Among the exploited biotechnological options, this latter beverage could be considered as a promising novel functional food.

The phenolic composition and antioxidant activity of several monovarietal extra virgin olive oils used as blenders for the production of Collina di Brindisi protected designation of origin (PDO) oil, produced between December 2008 and January 2009 using two-phases or three-phases extraction system, were evaluated and compared with other manufacturer products designated as PDO. Oils were taken from the most representative ones industrial oil mills in the PDO geographical area. The parameters assessed were free acidity, peroxide value, K 232 and K 270 indices, organoleptic characteristics, total phenolic content (TPC), phenolic profile, and antioxidant activity coefficient (AAC). The phenolic contents and profiles of the monovarietal oils showed remarkable differences with respect to PDO oils. The variables that exerted a major influence on phenols concentration were the maturity degree of olives (December>January), followed by the extraction system (two-phase>three-phase), and place of growing. The Pearson r correlation index showed that AAC was positively correlated with TPC, p-coumarate, and 3,4-DHPEA-EA, and negatively correlated with peroxide value. Practical applications: The results provide detailed information about: (i) the phenolic composition and the AAC of several monovarietal extra virgin olive oils used as blenders for the production of a PDO oil; (ii) the impact of genetic variability, place of growing, olive maturity degree, and extraction technology on oil phenol compounds; and (iii) the relationships among each phenolic compound and AAC, and their potential utilization as analytical index of antioxidant activity. It is important to study the phenolic compounds and antioxidant activity of monovarietal extra virgin olive oils used to produce PDO oil and to compare with the relative PDO samples in order to define a possible analytical tool able to verify what is stated in the label for consumer information and protection.

This article appeared in a journal published by Elsevier. The attached copy is furnished to the author for internal non-commercial research and education use, including for instruction at the authors institution and sharing with colleagues. Other uses, including reproduction and distribution, or selling or licensing copies, or posting to personal, institutional or third party websites are prohibited. In most cases authors are permitted to post their version of the article (e.g. in Word or Tex form) to their personal website or institutional repository. Authors requiring further information regarding Elsevier's archiving and manuscript policies are encouraged to visit: http://www.elsevier.com/copyright a b s t r a c t Spaghetti were produced in a pilot plant from semolina and semolina blended with increasing amounts of barley flour. According to size exclusion high-performance liquid chromatography (SE-HPLC), barley proteins interact with semolina proteins during pasta making, forming polymers of high molecular weight. Of these, the unextractable polymeric proteins (UPP) were at significantly higher concentrations than in spaghetti made from semolina. The decrease of both S–S bonds and –SH free groups in barley semolina spaghetti, with respect to that made of semolina, suggested that polymerisation among the different classes of proteins involves a new bonding arrangement. Due to b-glucan hydrophilicity and competition with starch for water, the replacement of increasing amounts of semolina with barley flour was able to increase the optimal cooking time. The sensory properties of composite spaghetti were judged as better than the control because of the higher firmness and the lower bulkiness and stickiness of the former.