I. Kandarakis

Evolution of microbial populations during traditional Feta cheese manufacture and ripening.

In three different dairies (A, B and C) located in Peloponess region (Southern Greece), traditional Feta cheese trials took place February to March using mixtures of sheeps and goats milk. Only small variations in the evolution of microbial groups were observed during the whole ripening period. The main groups, such as thermophilic cocci, mesophilic lactococci, thermophilic lactobacilli, nonstarter lactic acid bacteria (NSLAB), presumptive Leuconostoc, enterococci and micrococci, reached their highest levels during the first 16 days, and then declined approximately 1-2 log units until the end of ripening. The remaining groups investigated, comprising yeasts, coliforms and Escherichia coli, were highest at day 4. The yeasts remained constant, while coliforms and E. coli decreased sharply and were not detectable after 120 days of ripening. A number of 146 isolates (dairy A) taken from all stages of the manufacturing and ripening process were purified and studied. Lactobacillus plantarum (58/146) and isolates of related species Lactobacillus pentosus and Lactobacillus paraplantarum (16/146) were the most common microorganisms found during cheese ripening. Streptococcus thermophilus (23/146) and Lactobacillus delbrueckii subsp. bulgaricus (20/146) were detected in high levels up to 20 days, and then gradually reduced. Enterococcus faecium (29/146) was found in all manufacturing and ripening stages.

Cheese whey as a renewable substrate for microbial lipid and biomass production by Zygomycetes

Three Zygomycetes, Mortierella isabellina, Thamnidium elegans and Mucor sp., were tested for their ability of producing biomass and lipid‐containing γ‐linolenic acid (GLA) during their cultivation on cheese whey. M. isabellina consumed all of the available lactose and a significant amount of the available protein. On the contrary, the two other fungi seemed incapable of consuming lactose after protein exhaustion. In the second series of experiments, for M. isabellina a supplementary quantity of lactose was added into the medium in order to increase the C/N ratio and hence to increase the production of fat. In the case of T. elegans and Mucor sp., a supplementary quantity of ammonium sulfate was added in order to favor the consumption of lactose and the production of biomass. Indeed, enhancement of lipid production was observed for M. isabellina and biomass production for T. elegans and Mucor sp.. Fatty acid analysis of the microbial lipid showed a composition that presented non‐negligible changes in relation with the age of the culture and the C/N molar ratio of the medium. Further analysis of the fat showed that the quantity of neutral lipids was the more abundant. The fatty acid composition of neutral lipids resembled to that of total lipids. Phospholipids were the more unsaturated fraction for Mucor sp. and M. isabellina. GLA was synthesized in all trials but its concentration presented differences related with the utilized strains and the fermentation time. Growth of M. isabellina on lactose‐supplemented whey resulted in a maximum GLA production of 301 mg/L.

Effect of draining temperature on the biochemical characteristics of Feta cheese

Two different draining temperatures, 15 and 21°C were applied to five Feta cheese curds made with different starters, containing mesophilic or thermophilic strains or mixtures of them. After 20 h of draining, the pH of curds made with thermophilic starters ranged from 5.28 to 5.49. The draining temperature significantly affected (P<0.05) the pH and the total solids of the cheeses. The inclusion of whey proteins in the cheese curd due to the insufficient draining of cheeses at 15°C, resulted in higher water-soluble nitrogen (WSN), as % of total nitrogen content. Free amino acid contents were significantly affected (P<0.05) by the draining temperature and by the presence of thermophilic lactobacilli in the starter mixture. Draining temperature also significantly affected (P<0.05) residual αs- and β-casein and the RP-HPLC profiles of the WSN. The C2:0 to C8:0 free fatty acids, hardness (kg) and fracturability (kg), as well as the total organoleptic scores, were significantly (P<0.05) higher in feta drained at 21°C.

Effect of high-pressure treatment at various temperatures on indigenous proteolytic enzymes and whey protein denaturation in bovine milk

The objective of the present study was to determine the effect of high pressure (HP) processing (200, 450 and 650 MPa) at various temperatures (20, 40 and 55 degrees C) on the total plasmin plus plasminogen-derived activity (PL), plasminogen activator(s) (PA) and cathepsin D activities and on denaturation of major whey proteins in bovine milk. Data indicated that transfer of both PL and PA from the casein micelles to milk serum occurred at all pressures utilized at room temperature (20 degrees C). In addition to the transfer of PL and PA from micelles, there were reductions in activities of PL (16-18%) and PA (38-62%) for the pressures 450 and 650 MPa, at room temperature. There were synergistic negative effects between pressure and temperature on residual PL activity at 450 and 650 MPa and on residual PA activity only at 450 MPa. Cathepsin D activity in the acid whey from HP-treated milk was in general baroresistant at room temperature. The residual activity of cathepsin D decreased significantly at 650 MPa and 40 degrees C and at the pressures 450 and 650 MPa at 55 degrees C. Synergistic negative effects on the amount of native beta-lactoglobulin were observed at 450 and 650 MPa and on the amount of native alpha-lactalbumin at 650 MPa. There were significant correlations between enzymatic activities (PL, PA and cathepsin D) and the residual native beta-lactoglobulin and alpha-lactalbumin in bovine milk. In conclusion, HP significantly affected the activity of indigenous proteolytic enzymes and whey protein denaturation in bovine milk. Reduction in activity of indigenous enzymes (PL, PA and cathepsin D) and transfer of PL and PA from the casein to milk serum induced by HP is expected to have a profound effect on cheese yield, proteolysis during cheese ripening and quality of UHT milk during storage.

Nitrogenous fractions during the manufacture of whey protein concentrates from Feta cheese whey

Abstract Samples taken from different manufacturing stages of Feta cheese whey protein concentrates (WPCs), with 65 and 35% total protein content, were analyzed by chemical methods and by reversed-phase HPLC and by size-exclusion chromatography. Mean total protein content of Feta cheese whey was 1.37%. 22% of total nitrogen (TN) was non-protein N (NPN) and 12% was proteose–peptone N (PPN). The mean native N (NN), NPN and PPN percentages of WPC 65 and 35%, were 63 and 71%, 10 and 14%, 0.08 and 0.08%, respectively. According to reversed-phase HPLC data, the percentages of β -Lg and α -La and their ratio were high; 56.5 and 59% of native proteins of whey and of WPC powders, respectively, were β -Lg. The respective α -La percentages were 13.1 and 14.1% and the respective caseino-macropeptide (CMP) percentages were 12.9 and 15.3%. Apart from ultrafiltration, the most critical stage for nitrogenous fraction composition was the evaporation process before spray drying.

Purification and characterization of chymosin and pepsin from kid

The objective of this work was to study the characteristics of the gastric aspartic proteinases chymosin and pepsin which are constituents of the kid rennet. The two enzymes were extracted from abomasal tissue of one kid from a local indigenous breed, separated from each other by DEAE-cellulose chromatography and then were purified by gel filtration and anion-exchange chromatography. The molecular weights of the purified kid chymosin and pepsin as determined by gel filtration were 36 kDa and 40 kDa respectively. The isoelectric point of kid chymosin was as multiple forms of 3-6 zones at pH 4.6-5.1, while that of kid pepsin was at pH < or =3.0. Kid pepsin contained 0.37 molecules phosphorous per molecule and was totally inhibited by 5 muM pepstatin A, being more sensitive than kid chymosin. Both enzymes were almost equally as proteolytic as calf chymosin on total casein at pH 5.6. Kid pepsin activity was more pH and temperature dependent than kid chymosin activity. In comparison with the calf chymosin temperature sensitivity, the order of increased sensitivity was: calf chymosin