Compositional,biochemical and textural changes during ripening of Tulum cheese made with different coagulants

In the present study, biochemical, chemical and texture changes in Tulum cheeses made using calf rennet and microbial rennets (Aspergillus niger protease and Rhizomucor miehei protease) were compared during ripening for up to 90 days. A total of 15 free fatty acids (FFAs) were detected in the cheese samples. The peroxide values (PV) of the cheeses increased significantly (P < 0.05) during ripening and the cheese made with calf rennet had the highest PV. Proteolysis in the cheeses increased as the ripening time increased. α_s1‐casein and β‐casein degradation was higher in cheeses manufactured with R. miehei protease. Cheeses made with calf rennet were significantly (P < 0.05) harder, more adhesive, more cohesive and more resilient than those made with microbial rennet.     

Ripening of Camembert-type cheese made from caprine milk using calf rennet or kid rennet as coagulant

Camembert-type cheese was made from caprine milk using either calf rennet or kid 'Grandine' rennet as coagulant. The pH of all cheeses increased throughout ripening and levels of pH 4.6-soluble nitrogen increased from 8.1 to 18.2% of total nitrogen (TN) and from 6.9 to 20% TN for the cheeses made using calf rennet and kid rennet, respectively. Degradation of β-casein, measured by urea–polyacrylamide gel electrophoresis, and total and free amino acids were greater in the cheese made using kid rennet. Production of peptides, analysed by high performance liquid chromatography (HPLC), was slightly more extensive in the Camembert-type cheese made using calf rennet as coagulant. In general, a higher degree of proteolysis was found in Camembert-type cheese made from caprine milk using kid rennet than in cheese made using calf rennet as coagulant.     

酪蛋白降解对牦牛乳硬质干酪苦味的影响

针对牦牛乳硬质干酪的苦味缺陷,分别以小牛皱胃酶、微生物凝乳酶和木瓜蛋白酶制作的牦牛乳硬质干酪为研究对象,利用尿素聚丙烯酰胺凝胶电泳,研究牦牛乳硬质干酪pH 4.6水不溶性酪蛋白的降解程度,且对成熟过程中的牦牛乳硬质干酪苦味进行感官评价,探究牦牛乳硬质干酪pH 4.6水不溶性酪蛋白降解对其苦味的影响。结果表明：牦牛乳硬质干酪在成熟期间酪蛋白发生了明显的降解,且αs-酪蛋白均比β-酪蛋白降解速率快。经尿素聚丙烯酰胺凝胶电泳分离后,发现木瓜蛋白酶制作的牦牛乳硬质干酪pH 4.6水不溶性酪蛋白在Pre-αs-酪蛋白区域有较强的蛋白带。木瓜蛋白酶制作的牦牛乳硬质干酪pH 4.6水不溶性酪蛋白中αs-酪蛋白和β-酪蛋白降解程度均显著或极显著高于微生物凝乳酶和小牛皱胃酶制作的牦牛乳硬质干酪（P＜0.05或P＜0.01）,木瓜蛋白酶制作的牦牛乳硬质干酪的苦味值极显著高于微生物凝乳酶和小牛皱胃酶制作的牦牛乳硬质干酪的苦味值（P＜0.01）,通过主成分分析得出3 种凝乳酶制作牦牛乳硬质干酪的苦味值和未降解β-酪蛋白和αs-酪蛋白含量成极显著负相关。这为控制牦牛乳硬质干酪品质提供了理论参考。     

Proteolysis during the ripening of goats’ milk cheese made with plant coagulant or calf rennet

Powdered plant coagulant (PPC) obtained from the cardoon (Cynara cardunculus) was compared with calf rennet (CR) for the manufacture of goats’ milk cheese, by determining difference in the proteolysis throughout ripening. There were no substantial differences between the compositions of cheeses made using the two types of coagulants. However, cheeses manufactured with PPC exhibited higher levels of pH 4.6-SN than cheese made using CR. The extent of breakdown of α_s-casein, as measured by urea-PAGE, was greater in cheese made using PPC than cheese made using CR. The formation of hydrophobic peptides and the ratio of hydrophobic/hydrophilic peptides throughout the ripening were higher in cheeses made with PPC than in cheeses made with CR. Principal component analysis (PCA) of peak heights of RP-HPLC peptide profiles of the ethanol-soluble and ethanol-insoluble fractions distributed the samples according to the coagulant used in their manufacture. Quantitative differences in several peptides were evident between the two types of cheese.     

MILK CLOTTING AND CHEESE MAKING PROPERTIES OF A CHYMOSIN-LIKE ENZYME FROM HARP SEAL MUCOSA

Pepsin A was isolated from the gastric mucosa of two year old harp seal (Pagophilus groenlandicus). Seal pepsin A has a relatively high CU/PU ratio of 0.074, although it is lower than that of calf chymosin (0.170). Equivalent milk clotting units of chymosin and seal pepsin A catalyzed formation of the same amount of nonprotein nitrogen (NPN) when incubated with 2% casein and for both there was no appreciable increase in NPN formation in the second hour of incubation. Seal pepsin A was similar to chymosin in clotting milk substrate with respect to the influence of pH, dilution, calcium chloride and temperature. Cheddar cheeses prepared with either calf rennet or pepsin A as coagulating agent were similar in yield, chemical composition and taste as judged by sensory preference tests. Estimation of protein degradation in the aged cheeses by ultraviolet absorption of citrate-HC1 extracts, gel filtration chromatography of protein soluble in 6 N urea, and determination of amino acids indicated there was slightly more protein degradation in cheese prepared with calf rennet after 30 weeks aging. Sal pepsin A appears to make a major contribution to the excellent cheese-making characteristic of crude seal pepsin(s). However, other components of the seal mucosa extract are probably responsible for the accelerated aging of Cheddar cheese noted in previous reports.     

Evaluation of Thiol Activated Proteases from Clam Viscera as a Rennet Substitute for Cheese-Making

Clam rennet, which is a crude enzyme preparation of cathepsin B-like protease from clam viscera was characterized and compared to porcine pepsin and calf rennet for its suitability as a milk coagulant in cheesemaking. Clam rennet was more proteolytic and produced a softer curd than the other two coagulants. However, influences of the pH and temperature on milk clotting with clam rennet were very similar to those of calf rennet. The cheddar cheese made from clam rennet was not inferior to the Cheddar cheese made from calf rennet. Quality enhancement occurred despite the view that high ratio of proteolytic to clotting activity is generally considered to be unfavorable for cheese-making. The higher proteolytic activity appeared to accelerate the ripening process. A small yield loss occurred as a result of excessive proteolysis during cheese-making.     

Characterization and Cheese-Making Properties of Rennet-Like Enzyme Produced by a Local Algerian Isolate of Aspergillus niger

An ochratoxin free extracellular acid protease was produced by solid state cultivation of Aspergillus niger FFB1. The purified enzyme (48.7 kDa) showed an optimal milk clotting activity at pH 5.5 and 45°C in the presence of 0.01 M CaCl₂. The enzyme was stable at least 24 h at 35°C in the pH range of 5.5–7.0. Thermal denaturation started above 45°C. Fresh cheese manufactured with reconstituted cow milk and the purified enzyme showed similar basic characteristics (pH 4.5, acid taste, white color) as marketed cheeses obtained with calf rennet. This emphasizes the value of exploiting local biological resources for value added food processing in developing countries.     

Effect of artisanal kid rennet paste on lipolysis in semi-hard goat cheese

This study examined the use of hygienised kid rennet pastes in model cheese systems and also in goat milk semi-hard cheeses to promote lipolysis. The results obtained indicated that the use of rennet paste caused greater lipolysis and increased, mostly, the short-chain free fatty acid (FFA) content. The model systems made with whole goat’s milk using rennet paste and commercial rennet mixture exhibited a higher FFA content than did the rennet paste-free controls (31,600 vs. 25,600 μmol/kg cheese). For the pilot cheeses made with bovine rennet and rennet paste mixture, the increase in FFA level after 45 days of ripening compared with the cheeses prepared only with commercial rennet was as much as 6600 (μmol/kg cheese) and the increase in the butyric acid content was also 1650 (μmol/kg cheese). Moreover, after 15 days of ripening, industrially prepared cheeses made with rennet paste exhibited greater levels of FFA than did the cheeses made with commercial rennet (11,500 μmol/kg at 45 days of ripening). Their flavour was stronger and the organoleptic characteristics were better accepted, which implies less ripening time for commercial cheese manufacture.     

Comparison of buffalo cottage cheese made from aqueous extract of Withania coagulans with commercial calf rennet

An aqueous extract of Withania coagulans was used to prepare cottage cheese from buffalo milk and its quality attributes were compared with cheese made from commercial rennet. Both cheeses contain satisfactory ranges of 49.6–54.7% moisture, 21.3–24.3% fat and 21.4–23.6% protein. The type of coagulant had no significant effect on acidity, protein and ash contents of both the cheeses. W. coagulans cheese showed a significantly (P < 0.05%) higher pH and moisture contents. Similarly, no marked differences were observed in their organoleptic evaluation, actual and theoretical yield. These results supported the fact that W. coagulans is a promising rennet substitute for cottage cheesemaking.     

Characteristics of Miniature Cheddar‐Type Cheese Made by Microbial Rennet from Bacillus amyloliquefaciens: A Comparison with Commercial Calf Rennet

Miniature Cheddar‐type cheeses were produced using microbial rennet from Bacillus amyloliquefaciens (milk‐clotting enzyme [MCE]) or calf rennet (CAR). With the exception of pH, there were no significant differences in gross composition between MCE‐cheese (MCE‐C) and CAR‐cheese (CAR‐C). The pH value of CAR‐C was significantly higher than that of MCE‐C at 40 and 60 d of ripening. The total nitrogen content of the pH 4.6‐soluble fraction obtained from MCE‐C was higher than that obtained from CAR‐C. However, nitrogen content of the 12% TCA‐soluble fraction was similar between CAR‐C and MCE‐C. The extent of α_s1‐casein and β‐casein hydrolysis, measured by urea‐PAGE, was similar in both cheese samples. The hydrolysis of β‐casein was lower than that of α_s1‐casein. Different reverse phase‐high‐performance liquid chromatography peptide profiles of ethanol‐soluble and ethanol‐insoluble fractions were obtained from CAR‐C and MCE‐C. The peptide content in the 2 cheese samples increased throughout ripening; the ratio of hydrophobic to hydrophilic peptides was lower in MCE‐C than in CAR‐C. Compared with CAR‐C, MCE‐C was softer as a result of higher protein hydrolysis. Microbial rennet from B. amyloliquefaciens contributed to higher proteolytic rates, which reduced ripening time.     

Exogenous enzymes in dairy technology: acidic proteases from processing discards of shrimp Pleoticus muelleri and their use as milk‐clotting enzymes for cheese manufacture

Argentina is the seventh largest world producer of cheese. Owing to the profitability of this industry, more than a thousand of dairy companies and farms exclusively produce cheeses, and, thus, this activity utilises 42% of total raw milk production of the country. Due to the increase in cheese demand, and because calf rennet is relatively expensive, this has motivated the study of new sources of proteases due to the potential use in biotechnological processes. Previously, crustacean proteases have been considered as good rennet substitutes for milk coagulation during the cheese making process. The aim of this study was to evaluate and characterise acidic proteases obtained from Pleoticus muelleri discards and their proteolytic capability as well as their potential use as a milk‐clotting agent. The activity of aspartic proteases and the satisfactory acceptation by the taste panel for cheese made with shrimp's enzymes suggest that P. muelleri proteins offer a high potential for use in dairy biotechnological processes with potential use as milk‐clotting agents for cheese making.     

SENSORY CHARACTERISTICS OF EWE MILK CHEESE MADE WITH THREE TYPES OF COAGULANT: CALF RENNET, POWDERED VEGETABLE COAGULANT AND CRUDE AQUEOUS EXTRACT FROM CYNARA CARDUNCULUS

This article reports on a study of the sensorial characteristics of ewe milk cheese (Los Pedroches) made with animal rennet and two types of coagulant obtained from the cardoon Cynara cardunculus. The cheeses made with rennet displayed a less odor intensity: pungent odor and acid odor; a less acid taste; and a slightly lighter color. They were harder and firmer, but less creamy than those made with the vegetable coagulant (powdered vegetable coagulant [PVC] and crude aqueous extract [CAE] from C. cardunculus). However, the cheeses made with PVC showed organoleptic characteristics very similar to the cheeses made with CAE from C. cardunculus. After 90 days' ripening, the cheeses made with vegetable coagulant displayed a slightly more bitter taste than those made with rennet. In general terms, increased ripening time prompted the increased scores for most of the sensory attributes studied.     

Biochemical patterns in ovine cheese: Influence of probiotic strains

This study was undertaken to evaluate the effect of lamb rennet paste containing probiotic strains on proteolysis, lipolysis, and glycolysis of ovine cheese manufactured with starter cultures. Cheeses included control cheese made with rennet paste, cheese made with rennet paste containing Lactobacillus acidophilus culture (LA-5), and cheese made with rennet paste containing a mix of Bifidobacterium lactis (BB-12) and Bifidobacterium longum (BB-46). Cheeses were sampled at 1, 7, 15, and 30 d of ripening. Starter cultures coupled with probiotics strains contained in rennet paste affected the acidification and coagulation phases leading to the lowest pH in curd and cheese containing probiotics during ripening. As consequence, maturing cheese profiles were different among cheese treatments. Cheeses produced using rennet paste containing probiotics displayed higher percentages of α_S1-I-casein fraction than traditional cheese up to 15 d of ripening. This result could be an outcome of the greater hydrolysis of α-casein fraction, attributed to higher activity of the residual chymosin. Further evidence for this trend is available in chromatograms of water-soluble nitrogen fractions, which indicated a more complex profile in cheeses made using lamb paste containing probiotics versus traditional cheese. Differences can be observed for the peaks eluted in the highly hydrophobic zone being higher in cheeses containing probiotics. The proteolytic activity of probiotic bacteria led to increased accumulation of free amino acids. Their concentrations in cheese made with rennet paste containing Lb. acidophilus culture and cheese made with rennet paste containing a mix of B. lactis and B. longum were approximately 2.5 and 3.0 times higher, respectively, than in traditional cheese. Principal component analysis showed a more intense lipolysis in terms of both free fatty acids and conjugated linoleic acid content in probiotic cheeses; in particular, the lipolytic pattern of cheeses containing Lb. acidophilus is distinguished from the other cheeses on the basis of highest content of health-promoting molecules. The metabolic activity of the cheese microflora was also monitored by measuring acetic, lactic, and citric acids during cheese ripening. Cheese acceptability was expressed for color, smell, taste, and texture perceived during cheese consumption. Use of probiotics in trial cheeses did not adversely affect preference or acceptability; in fact, panelists scored probiotic cheeses higher in preference over traditional cheese, albeit not significantly.     

Behavior of Enteropathogenic Escherichia coli in Camembert Cheese Made From Ultrafiltered Milk

Skimmilk retentates from ultrafiltration (UF) were used in combination with concentrated cream (67% milk fat) to prepare a liquid precheese mixture that with the addition of rennet, lactic starter culture and a mold spore preparation of Penicillium candidum was transformed readily into Camembert cheese upon ripening. Yield increases from the retention of soluble milk protein, closer weight control tolerances for individual cheeses and reduction in rennet in comparison to the conventional Camembert process were observed. The behavior of enteropathogenic E. coli (ECC) serotypes was demonstrated in the Camembert cheese made from UF processes. Relationships were determined in UF Camembert as to microbial type and numbers used to inoculate the precheese, and as to site of survival and growth of different EEC serotypes. Differences which were observed in the physicochemical properties between conventional and UF cheesemilk mixtures predisposed the UF Camembert cheeses to greater E. coli survival and growth than in the Camembert cheeses made conventionally.     

Evolution of lipolysis during the ripening of traditional Feta cheese

Lipolysis was studied during ripening of traditional Feta cheese produced in two small dairies, A and B. The cheeses were made from a thermized mixture of ewes’/goats’ milk by using yoghurt as starter and artisanal rennet from lambs’ and kids’ abomasa (cheese A) or mixed artisanal rennet with calf rennet (cheese B).The acid degree value and the free fatty acids (FFA) contents in both cheeses increased sharply up to 18 d (pre-ripening period at 15 °C) and continued to increase throughout ripening. In both mature cheeses, acetic acid was found at high levels (13–18% of the total FFAs). However, except for this, all FFA contents differed significantly (P < 0.05) between the two cheeses throughout ripening. The levels of individual and total C2:0–C8:0, C10:0–C14:0 and C16:0–C18:2 fatty acids were significantly higher (P < 0.05) in cheese A than in cheese B. Presumably the difference, especially in the C2:0–C8:0 content, was due mainly to the type of the rennet used. Butyric acid was the dominant FFA in cheese A (20% of the total FFAs at 120 d), while the most abundant FFAs in cheese B were capric (18%) and lauric acid (18%). In general, the lipolysis degree of the two cheeses was higher than those reported for the industrially-made Feta cheese.In organoleptic evaluation, cheese A had a piquant taste that was attributed to its high content of butyric acid and showed a significantly (P < 0.05) higher total score than cheese B.     

Influence of rennet milk-clotting activity on the proteolytic and sensory characteristics of an ovine cheese

Roncal cheese (regulated by an Apellation of Origin) is a traditional hard cheese manufactured from raw ewe's milk in the region of Navarre in Spain. Roncal cheeses, manufactured using two lamb rennets with different milk-clotting activity levels, were evaluated to compare their chemical, proteolytic, and sensory characteristics. A preliminary study of samples of lamb rennets indicated that a large proportion of such rennets did not fulfil current microbiological requirements and likewise revealed considerable variation in the milk-clotting activity of the samples examined. Trends in the overall physicochemical parameter values (pH, dry matter, fat, and protein) were similar in both cheese batches. Proteolysis of the nitrogen fractions was observed to take place at a faster rate in the cheeses made using the rennet with the higher milk-clotting activity (soluble nitrogen, non-protein nitrogen, and amino acid nitrogen values around 13–20% higher than in the cheeses made using the rennet with the lower milk-clotting activity after 180 days of ripening). Urea-PAGE electrophoretic analysis of the caseins from the cheeses manufactured using both types of rennet showed that the β-caseins were less susceptible to proteolysis than the α_s-caseins. The effect of the different milk-clotting activity levels was most pronounced on the α_s-caseins, in which the rennet with the higher milk-clotting activity gave higher breakdown. Nevertheless, the differences in the proteolysis rates did not yield any appreciable sensory differences.     

Effects of coagulant type on the physicochemical and organoleptic properties of Kashar cheese

Kashar cheeses were manufactured using different coagulants (calf rennet, chymosin derived by fermentation and proteases from Rhizomucor miehei and Cryphonectria parasitica) and ripened for 90 days. Use of different coagulants did not influence the dry matter, fat, protein, salt, pH, titratable acidity, total free fatty acids and texture profile analyses. The levels of water‐soluble nitrogen, 12% trichloroacetic acid‐soluble nitrogen, and for 5% phosphotungstic acid‐soluble nitrogen, the sensory properties were significantly influenced by the use of different coagulants. β‐casein was more hydrolysed in the cheese manufactured using protease from Cryphonectria parasitica than the other cheeses during 90 d of ripening.     

Influence of lamb rennet paste on composition and proteolysis during ripening of Pecorino foggiano cheese

Rennet pastes produced by lambs subjected to three different feeding systems (mother suckling [MS], artificial rearing [AR], and artificial rearing with Lactobacillus acidophilus supplementation [ARLB]) and slaughtered at two different ages (20 and 40 d) were used for the manufacture of Pecorino foggiano cheese. Composition and proteolysis during ripening of Pecorino foggiano cheese (four replicates batches) were analyzed. Proteolysis was greater in cheeses made with rennet pastes from lambs slaughtered at 20 d, as shown by analysis of nitrogen fractions (water-soluble N and proteose peptones). Supplementation of milk substitute with L. acidophilus may have influenced the growth dynamics of lactic acid bacteria in the rennet pastes, with positive effects on levels of lactobacilli in cheese at the beginning of the ripening time. Lower pH values in ARLB cheese during ripening, together with higher cell loads, suggest that supplementation of milk replacer with L. acidophilus resulted in higher proteolytic activity, as also confirmed by the composition of the pH 4.6—insoluble nitrogen fraction. No differences were found in total concentration of free amino acids among the experimental cheeses; phenylalanine, isoleucine, leucine, lysine were found at the highest levels. The addition of probiotic bacteria to milk substitute in lamb rearing appears to give good-quality lamb rennet paste.     

Casein breakdown during ripening of Idiazabal cheese: influence of starter and rennet type

The objective of this work was to determine the effect of starter and rennet type on casein breakdown during Idiazabal cheese ripening. Four batches of cheeses were manufactured with two rennets, commercial calf rennet and artisanal lamb rennet, and the use of natural flora or a commercial starter. Electrophoretic analysis of cheese samples showed six bands identified as α_s1‐, α_s2 + β‐, α_s1‐I‐, γ₁‐, β‐I‐ and para‐κ‐casein. As expected, the casein breakdown during cheese ripening was considerably affected by rennet type and the use of a commercial starter. The artisanal lamb rennet produced a higher hydrolysis of casein fractions than the commercial calf rennet, probably owing to its high percentage of chymosin (around 78%). The effect of addition of starter on proteolysis was dependent on the casein fractions generated by artisanal lamb rennet or commercial calf rennet. © 2000 Society of Chemical Industry     

解淀粉芽孢杆菌GSBa-1凝乳酶在马苏里拉干酪中的应用

对分离自酒曲的1 株解淀粉芽孢杆菌GSBa-1发酵所产凝乳酶进行研究,该酶凝乳活力高而蛋白水解活力低,纯酶凝乳活力可达1.46×106 SU/g;使用该凝乳酶和商品凝乳酶制作马苏里拉干酪,并对干酪理化成分、成熟过程中pH值和微生物指标及干酪成熟前后质构特性、游离脂肪酸、可溶性蛋白、风味和干酪性能等指标进行对比分析。结果显示,理化成分上菌株凝乳酶与商品凝乳酶制作的干酪相接近（P＜0.05）。干酪在成熟过程中,发酵剂存活数先增加后减少,但其差异不大;菌株凝乳酶制作的干酪pH 4.6可溶性蛋白含量较多,干酪的游离氨基酸总量（76 mg/100 g）也高于商品凝乳酶制作的干酪游离氨基酸总量（55.3 mg/100 g）;菌株凝乳酶制作的干酪质构特性优于商品凝乳酶制作的干酪;电镜结果显示,菌株凝乳酶制作的干酪内部网状结构更充实;菌株凝乳酶具有稍强的蛋白水解活力,导致其制作的干酪风味物质种类多于商品凝乳酶制作的干酪,风味物质更加丰富。干酪样品的保形性和拉丝性实验测定结果显示,2 种凝乳酶制作的干酪性能差异不大（P＞0.05）;对2 种凝乳酶制作的干酪进行感官评定,其总评分相接近。以上结果表明,解淀粉芽孢杆菌GSBa-1凝乳酶在一定程度上可代替小牛凝乳酶应用于马苏里拉干酪的生产。