Proteolysis in miniature Cheddar-type cheeses made using blends of chymosin and Cynara cardunculus proteinases as coagulant

Miniature (20 g) Cheddar-type cheeses were manufactured using blends of Cynara cardunculus proteinases and chymosin as coagulant (100 : 0, 50 : 50, 25 : 75 and 0 : 100 C. cardunculus proteinases : chymosin). There were no substantial differences between the compositions of cheeses made using any of the four coagulant blends. Cheeses manufactured with coagulant blends containing C. cardunculus proteinases exhibited higher levels of pH 4.6-soluble nitrogen than cheese made using chymosin as coagulant. The extent of breakdown of α _s1 -casein, as measured by urea-polyacrylamide gel electrophoresis (urea-PAGE), was greater in cheeses made using coagulant preparations containing C. cardunculus proteinases as a constituent than in cheese made using 100% chymosin as coagulant. Different reverse-phase high-performance liquid chromatography (RP-HPLC) peptide profiles of the ethanol-soluble and -insoluble fractions were obtained for cheeses made using either C. cardunculus proteinases or chymosin as coagulant. Principal component analysis and hierarchical cluster analysis of RP-HPLC data confirmed that the inclusion of even small proportions (25%) of C. cardunculus proteinases with chymosin in the coagulant blend greatly altered the pattern and extent of proteolysis in miniature Cheddar-type cheeses.     

Effects of high pressure on proteolytic enzymes in cheese: relationship with the proteolysis of ewe milk cheese

Ewe milk cheeses were submitted to 200, 300, 400, and 500 MPa (2P to 5P) at 2 stages of ripening (after 1 and 15 d of manufacturing; P1 and P15). The high-pressure-treated cheeses showed a more important hydrolysis of β-casein than control and 2P1 cheeses. Degradation of α_s1-casein was more important in 3P1, 4P1, and P15 cheeses than control and 2P1 cheeses. The 5P1 cheeses exhibited the lowest degradation of α_s-caseins, probably as a consequence of the inactivation of residual chymosin. Treatment at 300 MPa applied on the first day of ripening increased the peptidolytic activity, accelerating the secondary proteolysis of cheeses. The 3P1 cheeses had extensive peptide degradation and the highest content of free amino acids. Treatments at 500 MPa, however, decelerated the proteolysis of cheeses due to a reduction of microbial population and inactivation of enzymes.     

Effect of lyophilisation,refrigerated storage and frozen storage on the coagulant activity and microbiological quality of Cynara cardunculus L. extracts

BACKGROUND: Cheese‐makers have traditionally kept vegetable coagulants refrigerated until use, even though little was known of their microbiological quality or coagulant activity during storage. This study aimed to assess the efficacy of lyophilisation, refrigerated storage and frozen storage of fresh vegetable extract as a means of standardising coagulant activity in terms of coagulation times, pH and microbiological quality. RESULTS: Neither the pH nor the coagulation time of lyophilised extracts was significantly modified during 1 year; however, changes were observed following frozen storage, and more notable following refrigerated storage. Lyophilisation of aqueous extracts prompted the destruction of most micro‐organisms; low counts initially noted for total mesophiles, lactic acid bacteria and yeasts disappeared during the first few days of storage, due to low water activity. There was a generalised decrease in micro‐organism counts during frozen storage. Refrigeration was found to be unsuitable for storing of cardoon extract; an increase of roughly 2 log unit counts was recorded in total mesophile, lactic acid bacteria, yeast and mould counts after 1 year of refrigerated storage. CONCLUSION: Refrigerated storage cannot be considered a suitable method for prolonged conservation of aqueous cardoon extract. Both lyophilisation and frozen storage of aqueous extracts proved ideal for prolonged storage of vegetable coagulant. Lyophilisation additionally had certain advantages over frozen storage. Copyright © 2008 Society of Chemical Industry     

Free amino acid content of goat's milk cheese made with animal rennet and plant coagulant

BACKGROUND: Enzymes present in the flowers of Cynara cardunculus (cyprosins) are used in the production of some traditional Spanish and Portuguese cheeses, replacing animal rennet. The aim of this work was to study the changes that take place in free amino acids during the ripening of a goat's milk cheese (Murcia al Vino) manufactured with plant coagulant (PC) or animal rennet (AR). RESULTS: The total free amino acid (TFAA) concentration increased during ripening, with Ile, Val, Ala, Phe, Gaba, Arg and Lys representing more than 50% of the TFAA content at 60 days in both types of cheese. The TFAA concentration was significantly higher in cheeses made with PC (854 mg 100 g^?1 total solids (TS)) than those made with AR (735 mg 100 g^?1 TS). The concentration of most free amino acids, especially His, Ser, Gln, Thr, Ala, Met and Ile, was higher in the PC cheese. CONCLUSION: Cheese made using PC as coagulant presented higher contents of free amino acid throughout the ripening period than cheese made using AR. Therefore we can conclude that the use of PC to produce Murcia al Vino goat's cheese would accelerate the ripening process as a result of increased cyprosin proteolytic activity. Copyright © 2011 Society of Chemical Industry     

热激处理菌种对奶酪品质的影响

为研究热激处理（63 ℃、20 min）发酵剂菌种（唾液链球菌嗜热亚种和保加利亚乳杆菌德氏亚种混合菌种）对奶酪成熟过程中品质特性的影响,对两种不同菌种所制作奶酪基本组成及成熟过程中的质构、感官、蛋白降解、风味物质等指标进行了测定。结果表明,热激处理菌种对奶酪基本组成影响不大,但热激处理菌种有利于奶酪成熟过程中蛋白质的降解,进而使奶酪的硬度和咀嚼性显著降低,也促使奶酪在成熟120 d时形成了更多的醛类、酮类、酯类等主要挥发性风味物质,同时奶酪的风味和组织状态的感官评价也较好。     

Characterization of Crude Esterase Activity from Two Plants Used in Cheese Making: Cynara cardunculus L. and Ficus carica L.

For centuries, extracts of Cynara cardunculus and the latex of Ficus carica have been used in cheesemaking, thanks to their proteases that clot milk casein. Since esterase activity is important to release odorous compounds in cheese, in this study we characterized the esterase activity of an extract of Cynara cardunculus and the latex of Ficus carica. The results, obtained by titration, spectrophotometric techniques, zymograms, and tests on agar plates, show the presence of esterase activity in the samples tested, with affinity towards the esters of short chain fatty acids, such as acetic, propionic, and butyric acid. In conclusion, this study provides a background information on esterases of two plants used in cheesemaking and represents a starting point for further investigations of their applications, not only in dairying but also in other industries involved in the processing and transformation of lipids.     

Enzymatic extract from flowers of Algerian spontaneous Cynara cardunculus: Milk‐clotting properties and use in the manufacture of a Camembert‐type cheese

The aim of this study was to assess the coagulant properties of crude extract from cardoon flowers (CECF) as a replacement for the imported coagulants used in the local Algerian cheese factories. Optimal coagulation activity of the extract was observed at pH 5 and 60 °C with a CaCl₂ concentration of 0.02 M. The electrophoretic pattern revealed that the crude extract contains mainly cardosins A and B. The study of interactions involved in the formation of milk gel coagulated by CECF showed that hydrogen forces, hydrophobic interactions and calcium bridges contributed highly to the formation of milk gel. Camembert‐type cheese obtained by CECF was judged by trained panellists and was found to be in agreement with standard specifications. The present study highlights the possibility of using Algerian CECF as a promising plant coagulant in cheesemaking for the dairy industry.     

发酵剂对牦牛乳硬质干酪成熟过程中生物胺的影响

乳酸菌产生物胺的能力具有菌株特异性,因此,为了探究不同种类发酵剂对牦牛乳硬质干酪中生物胺形成的影响,该试验利用高效液相色谱对3种不同发酵剂制作的硬质干酪成熟过程中生物胺进行了测定和分析。结果表明,嗜热和嗜温发酵剂牦牛乳硬质干酪中检测出2-苯乙胺、腐胺、尸胺、组胺和酪胺,混合发酵剂干酪中检测出腐胺、2-苯乙胺、尸胺和酪胺。各生物胺之间呈现正相关性。3种不同发酵剂干酪在1～6个月成熟过程中,其各生物胺整体呈现增加趋势,嗜热、嗜温和混合发酵剂干酪中总生物胺最高含量分别为(448.3±9.6)、(456.8±58.4)、(293±24.5)mg/kg。组胺和酪胺是2种毒性相对高的生物胺,嗜热发酵剂干酪中组胺和嗜温发酵剂干酪中酪胺最高,其最高含量分别为(20.8±7.9)、(92.9±6.7)mg/kg,混合发酵干酪中未检测出组胺,酪胺含量次之,3种不同发酵剂干酪中组胺、酪胺含量均低于推荐安全剂量50 mg/kg和100 mg/kg。这为合理选择发酵剂和控制干酪中生物胺形成提供了依据。     

Effect of starter cultures on properties of soft white cheese made from camel (Camelus dromedarius) milk

This experiment was conducted to investigate the effect of starter cultures on the physicochemical properties, texture, and consumer preferences of soft white cheese (SWC) made from camel (Camelus dromedarius) milk. The experiment was laid out in a completely randomized design with 5 treatments [starter cultures; i.e., 1 thermophilic (STI-12), 2 blended (RST-743 and XPL-2), and 2 mesophilic (R-707 and CHN-22) cultures]. Starter cultures STI-12 and RST-743 were inoculated at 37°C, whereas XPL-2, R-707, and CHN-22 were inoculated at 30°C. Camel milk inoculated using STI-12 and RST-743 cultures resulted in faster acidification than XPL-2, R-707, and CHN-22 cultures. Camel milk SWC made using STI-12 and CHN-22 cultures gave lower pH (4.54) and titratable acidity (0.59), respectively, whereas R-707 culture resulted in high cheese yield (13.44 g/100 g). In addition, high fat (20.91 g/100 g), protein (17.49 g/100 g), total solids (43.44 g/100 g), and ash (2.40 g/100 g) contents were recorded for SWC made from camel milk made using RST-743 culture. Instrumental analysis of cheese texture revealed differences in resistance to deformation in which camel milk SWC made using RST-743 culture gave higher firmness (3.20 N) and brittleness (3.12 N). However, no significant difference was observed among camel milk SWC adhesiveness made using different starter cultures. Consumer preference for appearance, aroma, taste, and overall acceptances of SWC were affected by inoculation of starter cultures. Considering curd firmness, cheese yield, compositional quality, and textures using STI-12, RST-743, and R-707, these cultures were found to be better for the manufacture of camel milk SWC.     

Effect of whey pH at drainage on physicochemical, biochemical, microbiological, and sensory properties of Mozzarella cheese made from buffalo milk during refrigerated storage

The objective of this research was to determine the effect of drainage pH on physicochemical, biochemical, microbiological and sensory properties of Mozzarella cheese made from buffalo milk during refrigerated storage. Four vats of cheese were made at 4 different whey drainage pH (6.2, 5.9, 5.6, and 5.2). Lower drainage pH caused higher pH 4.4-soluble N and pH 4.4-soluble N:total N. Interaction of drainage pH at d 1 and 30 of storage on all soluble nitrogen fractions was significant. Degradation of caseins in samples made at a drainage pH of 6.2 was lower than that of other cheese samples. The decreasing whey drainage pH significantly increased counts of thermophilic and mesophilic lactobacilli of the samples during refrigerated storage. No coliforms or Escherichia coli were detected in the cheeses. The average sensory property scores of all cheese samples were very close, and, as expected, storage time had a negative effect on all sensory scores.     

Proteolysis and texture changes of a Spanish soft cheese (‘Torta del Casar’) manufactured with raw ewe milk and vegetable rennet during ripening

Proteolysis and textural changes of the Spanish ewe raw milk soft cheese of the Protected Designation of Origin Torta del Casar were studied in four different stages of ripening, with 1, 30, 60 and 90 days. In general, proteolysis in Torta del Casar cheese was weak at 1 and 30 days and it was more intense between the 30–60 days of ripening. Soluble nitrogen non‐protein nitrogen, polypeptide N and free amino acids values significantly increased during cheese ripening. Protein and casein nitrogen decreased significantly after 60 days of ripening resulting in the increase of the other nitrogen fractions measured. Caseins changes determined by capillary zone electrophoresis showed that proteolysis of β‐casein occurred faster than αs1‐casein but the latter suffered higher proteolytic degradation at the end of ripening (day 90). This pattern of degradation of caseins is reversed in other cheeses made with animal rennet. Texture analysis showed that firmness and consistency decreased along ripening while adhesiveness increased. Highly significant correlations were found between textural parameters, residual caseins levels and nitrogen fractions during maturation, which shows the importance of proteolytic changes for an optimal texture formation.     

发酵剂用量对硬质蒙古干酪蛋白质、脂类水解和微观结构的影响

该文通过添加不同发酵剂用量制作了3种硬质蒙古干酪,并对硬质蒙古干酪的理化性质、蛋白质和脂肪水解程度、微观结构等进行了研究。结果表明,随着发酵剂用量增多,除D-乳酸含量外,其他理化指标均随发酵剂用量的增大而减小;粗蛋白质和粗脂肪的含量减少;pH 4.6-可溶性氮（SN）含量和12%三氯乙酸（TCA）-SN含量均增加,表明干酪中蛋白质和脂肪的水解程度增大;游离氨基酸和游离脂肪酸的含量增加,表明干酪的风味物质增多。随着发酵剂用量的增多,干酪酸化速度加快,质地相对松散。最终确定发酵剂使用量为1.0 g/L,在此条件下,脂肪和蛋白质的水解程度适中,游离氨基酸种类丰富,能够满足干酪风味化合物生成的需求。     

Proteolysis texture and microstructure of low‐fat Tulum cheese affected by exopolysaccharide‐producing cultures during ripening

The objective of the study was to determine the effects of exopolysaccharide (EPS)‐producing or non‐EPS‐producing starters on proteolysis, physical and microstructural characteristics of full‐fat or low‐fat Tulum cheeses during ripening. For this purpose, Tulum cheese was manufactured using full‐ or low‐fat milk with EPS‐producing and non‐EPS‐producing starter cultures. Chemical composition, proteolysis, texture profiles and microstructure of the cheeses were studied during 90 days of ripening. Urea‐PAGE of water‐insoluble and RP‐HPLC peptide profiles of water‐soluble fractions of the cheeses showed that the use of starters resulted in different degradation patterns in all cheeses during ripening. Although β‐casein exhibited similar degradation patterns in all cheeses, small differences are present in α_s1‐casein degradation during ripening. Reducing fat in Tulum cheese changed the RP‐HPLC peptide profile of the cheeses. The use of EPS‐producing cultures improved the textural characteristics and changed the microstructure and proteolysis of low‐fat Tulum cheese.     

Monitoring the chemical and textural changes during ripening of Iranian White cheese made with different concentrations of starter

The effect of the concentration of starter inoculated to milk on the composition, free tyrosine-tryptophan content, microstructure, opacity, and fracture stress of Iranian White cheese (IWC) was studied during 50 d of ripening in brine. Three treatments of cheese were made using 1-fold (IWC1S), 2-fold (IWC2S), and 4-fold (IWC4S) concentrations of a direct-to-vat mesophilic mixed culture containing Lactococcus lactis ssp. cremoris and Lactococcus lactis ssp. lactis as starter. As ripening progressed, moisture and protein contents of the treatments continuously decreased, whereas their total ash, salt, and salt in moisture contents increased. Fat content and pH of cheeses remained stable during ripening. The pH of cheese milk at the time of renneting, which decreased by increasing the concentration of starter (6.57, 6.49, and 6.29 for IWC1S, IWC2S, and IWC4S, respectively), significantly affected most of the chemical characteristics and opacity of cheese. Lower pH values at renneting decreased moisture and ash contents, whereas cheese protein content increased. The concentration of free tyrosine-tryptophan in curd increased at first 29 d but decreased between d 29 and 49 of aging. The changes observed in cheese whiteness followed the changes in moisture content of the treatments. As the concentration of starter inoculated to milk increased, the value of fracture stress at a given ripening time significantly decreased, leading to a less resistant body against applied stress. A similar trend was also observed for fracture strain during cheese ripening. The micrographs taken by scanning electron microscopy provided a meaningful explanation for decrease in the value of fracture stress. As the cheese ripening progressed or the concentration of starter increased, the surface area occupied by the protein fraction in cheese microstructure decreased, leading the way to lower the force-bearing component in cheese texture.     

Proteolysis, lipolysis, volatile compounds, texture, and flavor of Hispánico cheese made using frozen ewe milk curds pressed for different times

Hispánico cheese is manufactured in Spain from a mixture of cow and ewe milk. Production of ewe milk varies throughout the year, with a peak in spring and a valley in summer and autumn. To overcome this seasonal shortage, curd from spring ewe milk may be frozen and used for cheese manufacture some months later. In the present work, ewe milk curds pressed for 15, 60, or 120 min were held at −24°C for 4 mo, thawed, cut to 1-mm pieces, and mixed with fresh cow milk curd for the manufacture of experimental Hispánico cheeses. Control cheese was made from a mixture of pasteurized cow and ewe milk in the same (80:20) proportion. Cheeses, made in duplicate experiments, were analyzed throughout a 60-d ripening period. No significant differences between cheeses were found for lactic acid bacteria counts, dry matter content, hydrophilic peptides, 47 out of 68 vol.tile compounds, texture, and flavor characteristics. On the other hand, differences of minor practical significance between experimental and control cheeses, unrelated to the use of frozen ewe milk curd or the pressing time of ewe milk curd, were found for pH value, aminopeptidase activity, proteolysis, hydrophobic peptides, free amino acids, free fatty acids, and the remaining 21 vol.tile compounds. It may be concluded that the use of frozen ewe milk curd in the manufacture of Hispánico cheese does not alter its main characteristics.     

Effect of the lactoperoxidase system on the activity of mesophilic cheese starter cultures in goat milk

The effect of goats’ milk lactoperoxidase (LP) system on the activity of commercially available mesophilic cheese starter cultures was investigated. The growth and acid production of the starter cultures were measured at 2 h intervals for 8 h in goats’ milk kept at 30°C. Most of the starter cultures examined were found to be sensitive to the LP system, but varied in their susceptibility to inhibition. The activity of the mixed starter cultures CHN11, CHN22, CHN19, DCC240 and Flora Danica Normal was strongly inhibited by the LP system. However, the mixed starter culture LL 50C showed resistance to the LP system. The single strain culture Lactococcus lactis subsp. lactis NCDO 605 was inhibited by the LP system. However, the cultures Lactococcus lactis subsp. diacetylactis NCDO 176 and Leuconostoc mesenteroides subsp. cremoris ATCC 33313 were insensitive to the LP system. The results of this study indicate the need for routine screening of starter cultures for resistance to the LP system before using them for cheesemaking from goats’ milk preserved by the LP system.