Biochemical and microbiological characterization of artisan kid rennet extracts used for Cabrales cheese manufacture

Four samples of artisanal kid rennet extracts from four different Cabrales cheese producers were biochemically and microbiologically characterized. Most samples had a very acidic pH (around pH 4.0), which may condition their biochemical and microbial variables. The milk-clotting activity of the extracts was assessed using a Formagraph apparatus. The properties of these artisanal rennets were found to be comparable to a 1/10 dilution of 1:10,000 commercial calf rennet; their enzymatic potentials, measured with a semi-quantitative commercial system (API ZYM), showed only very slight differences. A large population of lactobacilli was found in all artisanal kid rennet samples, whereas coliforms, enterococci, staphylococci and leuconostocs were only occasionally encountered. Sixty-four representative colonies were classified by PCR amplification and the sequencing of a stretch of their 16S rDNA genes. Strains of Lactobacillus plantarum completely dominated one of the extracts. In all others samples, strains of this homofermentative species and of the heterofermentative Lactobacillus brevis were present in similar amounts.     

Short communication: Sensory profile of raw goat milk cheeses made with artisan kid rennet pastes from commercial-weight animals: Alternative to farmhouse goat cheeses

The loss of traditional kid rennet pastes in the Canary Islands (Spain), as in many other regions, is most likely due to the custom of using abomasa from very young animals killed below desirable commercial weight. In addition, the reasonable price of commercial rennets (CR) has resulted in the loss of typical sensory characteristics for most farmhouse raw goat milk cheeses, placing them at a disadvantage when local and international markets are full of different cheeses, often with aggressive marketing strategies. This paper analyzes the sensory characteristics of raw goat milk cheeses made with rennet pastes prepared from commercial kid abomasa in 2 ways: dried while full of ingested milk [full, commercial, artisan kid rennet (FCKR)], or dried after being emptied of ingested milk and refilled with raw goat milk [empty, commercial, artisan kid rennet (ECKR)]. This latter practice allows the use of empty abomasa, or abomasa with grass, soil, and so on. Sensory profiles of cheeses made with FCKR and ECKR rennets were compared with those made with CR by an expert panel (n = 7). The FCKR and ECKR cheeses had similar sensory profiles. Although scores for FCKR cheeses were somewhat higher than for ECKR cheeses, they were in the range found for traditional cheeses made with rennet prepared with abomasa from very young animals. The sensory profile of CR cheeses was very different. Almost 90% of consumer panelists (n = 90) preferred cheeses made with the experimental rennet pastes. These results demonstrate the possibility to prepare artisan rennet pastes from commercial-weight kids in an easy way for farmhouse cheese makers using local resources that would otherwise be destroyed in abattoirs.     

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     

Some properties of set yoghurt made from caprine milk and bovine–caprine milk mixtures fortified by ultrafiltration or the addition of skim milk powder

Set yoghurt was produced from caprine milk (A), 70% caprine-30% bovine (B) and 50% caprine–50% bovine milk (C) mixtures, and stored for 14 days at ±4°C. Two methods of fortification, namely ultrafiltration (UF) and the addition of bovine skim milk powder (SMP), were applied to the milk mixtures. Some chemical, physical, microbiological and sensory properties of the six samples were analysed on the 1st, 7th and 14th days of storage. The effects of milk type, concentration method and storage period on the physicochemical and microbiological properties of the samples were investigated statistically.     

凝乳酶对超滤浓缩乳生产Quark干酪的影响

采用每100g超滤乳中添加0、50、100、150、200、250μL六个水平的凝乳酶,研究了不同的凝乳酶添加量对Quark干酪组成、凝乳硬度、贮藏期产品感官品质和干酪中水溶性氮含量的影响。结果表明,当凝乳酶的添加量从0μL/100g增大到250μL/100g时,产品的水分含量上升了1·49%,粗产率和校正产率分别上升了1·42%和0·99%,固形物的回收率下降了3·5%,凝乳硬度从16·83g增大到40·84g,但干酪的苦味和水溶性氮含量,随着贮存期的延长和凝乳酶用量的增加而增大。      

Texture and flavour development in Ras cheese made from raw and pasteurised milk

Texture, proteolysis and flavour development in Ras cheeses made from raw or pasteurised milk with two different thermophilic lactic cultures were monitored during ripening. Results showed that at day 1 of manufacture, the moisture content and pH were lower in raw milk cheese than in pasteurised milk cheeses. Levels of water-soluble nitrogen, casein breakdown, free amino groups and free fatty acids were higher in cheese made from raw milk than in that made from pasteurised milk. Textural characteristics, such as hardness, cohesiveness and chewines, increased in all treatments during the first 60 days of ripening due to the reduction in the moisture level during the second stage of salting (dry salting during the first 60 days of ripening). Cheese made from raw milk received the highest texture and flavour scores by panellists.     

Quality evaluation of mozzarella cheese made from buffalo milk by using paneer booti (Withania coagulans) and calf rennet

This systematic research was conducted to assess the suitability of Withania coagulans for the preparation of mozzarella cheese from buffalo milk. The extract of dried berries of paneer booti (W. coagulans) was prepared in three buffers separately viz. Tris‐HCl, phosphate, NaCl and appropriate concentration of crude extracts were tested for their milk coagulating activity. NaCl solution with 0.85% strength was found to be suitable for milk coagulation without any objectionable taste and flavour. The results revealed that 15 μL crude enzyme extract per mL of milk was the optimum concentration for coagulation while 4.25 and 37 °C were the best levels of pH and incubation temperature for coagulation respectively. Mozzarella cheese prepared from crude enzyme extract was compared with the cheese prepared by calf rennet using acidification process. There was a nonsignificant difference among all tested parameters of both the coagulants. These findings support the suitability of an aqueous extract of W. coagulans for the commercial preparation of different types of mozzarella cheeses.     

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.     

Changes in texture,composition and sensory characteristics of Camembert cheese made from a mixture of goat milk and cow milk during ripening

The study was conducted to evaluate the nutritional, textural and sensory characteristics of Camembert cheese made from goat milk, cow milk and their blends. The type of milk used affected yield, physicochemical properties and fatty acid profile of the cheeses. The percentages of C6:0, C8:0 and C10:0 acids were higher in the cheeses containing ≥ 75% goat milk. The sensory evaluation results showed differences in texture, colour, aroma and flavour among the cheeses. Cheeses made with the mixture of cow and goat milk particularly 50% goat milk mixed with 50% cow milk (50G:50C) maintained particular positive nutritional characteristics especially with respect to the mineral contents, fatty acids and consumer acceptability.     

The effect of coagulant type on yield and sensory properties of semihard cheese from laboratory‐, pilot‐ and commercial‐scale productions

Using calf rennet or a commercial microbial rennet substitute derived from Rhizomucor miehei cheesemaking experiments were performed at laboratory and pilot scale, and at commercial scale in two industrial dairy plants during regular production. At all levels of scale, the solids transfer from milk to curd was significantly higher (0.50–1.19%) when using calf rennet. There were significant differences in levels of proteolysis during maturation and in levels of bitterness at 12 weeks of ripening between Gouda cheeses produced with calf rennet or with commercial rennet substitute at pilot and at commercial scale.     

Inclusion of microalgae in the caprine diet improves nutritional profile of milk and its Camembert cheese

The experiment was carried out to investigate the effects of supplementation with microalgae (Schizochytrium sp.) on the nutritional characteristics of milk and Camembert cheese from dairy goats. The yield and composition of milk and cheese and the sensory quality of cheese were not affected by the supplementation. As the level of supplementation increased, the concentration of docosahexaenoic acid and total n-3 fatty acids increased, whereas the ratio of n-6/n-3 and total saturated fatty acids decreased in the milk and cheese. In conclusion, supplemented goats, especially goats that received 35 g/head/day produced milk and cheese with better fatty acid composition for human consumption, without affecting the production and composition.     

Purification and characterization of a pregastric esterase from a hygienized kid rennet paste

Rennet pastes obtained by maceration of gastric tissues from suckling kids are used traditionally to produce some artisanal cheeses in Spain. Besides milk-clotting function, rennet pastes provide proteolytic activity and lipolytic system, essentially pregastric, necessary in the development of piquant flavor typical of these cheeses. A simple and reproducible procedure allows us to obtain a standardized rennet paste that posses the desired activity and is of good microbiological quality. Concomitantly, a kid pregastric esterase (KPGE) was purified to homogeneity. The purification procedure was based on an aqueous extract of hygienized rennet paste (HRP), which was chromatographed on DEAE-Sepharose Fast Flow then adsorbed on phenyl superose followed by a re-chromatography on the same column. The final enzymatic preparation, where the overall activity recovery was 3%, showed a molecular mass of 53 kDa. The highest activity was determined on p-nitrophenyl butyrate, but marked hydrolysis was also detected on beta-naphthyl caprylate. In contrast, low activity on tributyrin (substrate under emulsion form) was detected, thus confirming the esterase character of purified enzyme.     

Characterisation of Macedonian white‐brined cheese: Effect of raw or heat‐treated caprine milk

This research analysed the use of raw and pasteurised milk in the production of Macedonian white caprine milk cheese. Pasteurisation resulted in a considerable decrease in the nitrogen fractions, urea‐polyacrylamide gel electrophoresis of caseins, peptide profiles and volatiles during the maturation period. Forty‐five volatile components were detected, including twelve acids, fourteen esters, six ketones, three alcohols, four terpenes and six other compounds. It was deduced that pasteurisation of milk for the manufacture of artisanal white‐brined cheese delayed the evolution of volatiles, which are related to the unique flavour of the cheese.     

Changes during ripening in chemical composition,proteolysis, volatile composition and texture in Kashar cheese made using raw bovine,ovine or caprine milk

Kashar cheeses were manufactured from pure ovine (OV), bovine (BV) and caprine (CP) milk, and the chemical composition, cheese yield, proteolysis, hardness, meltability and volatile composition were studied during 90 days. Gross chemical composition, cheese yield and level of proteolysis were higher in OV cheeses than those of BV or CP cheeses. Glu, Val, Leu, Phe and Lys were the most abundant free amino acids (FAA) in the samples, and the concentrations of individual FAA were at the highest levels in OV cheeses with following BV and CP cheeses. Urea‐PAGE patterns and RP‐HPLC peptide profiles of the BV cheeses were completely different from the small ruminants’ milk cheeses (OV or CP). Higher and lower hardness and meltability values were observed in CP cheeses, respectively. OV cheeses resulted in higher levels of the major volatile compounds. In conclusion, the Kashar cheese made using OV milk can be recommended due to high meltability, proteolysis and volatiles.     

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.     

The fate of Listeria monocytogenes during the manufacture of Camembert-type cheese: A comparison between raw milk and milk treated with high hydrostatic pressure

Camembert-type cheese was produced from: raw bovine milk; raw milk inoculated with 2 or 4 log CFU/ml Listeria monocytogenes; raw milk inoculated with L. monocytogenes and subsequently pressure-treated at 500 MPa for 10 min at 20 °C; or uninoculated raw milk pressure-treated under these conditions. Cheeses produced from both pressure-treated milk and untreated milk had the typical composition, appearance and aroma of Camembert. Curd and cheese made from inoculated, untreated milk contained large numbers of L. monocytogenes throughout production. An initial inoculum of 1.95 log CFU/ml in milk increased to 4.52 log CFU/g in the curd and remained at a high level during ripening, with 3.85 log CFU/g in the final cheese. Pressure treatment inactivated L. monocytogenes in the raw milk at both inoculum levels and the pathogen was not detected in any of the final cheeses produced from pressure-treated milk. Therefore high pressure may be useful to inactivate L. monocytogenes in raw milk that is to be used for the production of soft, mould-ripened cheese.

Industrial relevance

This paper demonstrates the potential of high pressure (HP) for treatment of raw milk to be used in the manufacture of soft cheeses. HP treatment significantly reduced the level of Listeria monocytogenes in the raw milk and so allowed the production of safer non-thermally processed camembert-like soft cheese.     

Effect of buttermilk made from creams with different heat treatment histories on properties of rennet gels and model cheeses

Although many studies have reported negative effects on cheese properties resulting from the use of buttermilk in cheese milk, the cause of these effects has not been determined. In this study, buttermilk was manufactured from raw cream and pasteurized cream, as well as from a cream derived from pasteurized whole milk. Skim milks with the same heat treatments were also manufactured to be used as controls. Compositional analysis of the buttermilks revealed a pH 4.6-insoluble protein content approximately 10% lower than that of the skim milk counterparts. Milk fat globule membrane (MFGM) proteins remained soluble at pH 4.6 in raw cream buttermilk; however, when heat was applied to cream or whole milk before butter making, MFGM proteins precipitated with the caseins. Rennet gel characterization showed that MFGM material in the buttermilks decreased the firmness and increased the set-to-cut time of rennet gels, but this effect was amplified when pasteurized cream buttermilk was added to cheese milk. The microstructure of gels was studied, and it was observed that gel appearance was very different when pasteurized cream buttermilk was used, as opposed to raw cream buttermilk. Model cheeses manufactured with buttermilks tended to have a higher moisture content than cheeses made with skim milks, explaining the higher yields obtained with buttermilk. Superior retention of MFGM particles was observed in model cheeses made from pasteurized cream buttermilk compared with raw cream buttermilk. The results from this study show that pasteurization of cream and of whole milk modifies the surface of MFGM particles, and this may explain why buttermilk has poor coagulation properties and therefore yields rennet gels with texture defects.     

Production of hard cheese from caprine milk by the use of two types of probiotic cultures as adjuncts

Hard cheeses (Kefalotyri-like) were manufactured from caprine milk with yoghurt as a starter (A), and with its partial replacement with the probiotic adjuncts Lactobacillus rhamnosus LC 705 (B) and/or Lactobacillus paracasei ssp. paracasei DC 412 (C). Both adjuncts retarded the growth of enterococci, and the environment in cheese B did not favour the recovery of lactic acid bacteria (LAB) on Rogosa agar. However, better recovery of the LAB population on M17 agar from cheeses B and C made with adjuncts was recorded early in ripening, and this was accompanied by a greater decrease in pH. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of whole-cell protein demonstrated that cheese C, made with Lb. paracasei ssp. paracasei as adjunct, is a better vehicle for delivery of live probiotic cells (10 ⁷   cfu/g) to the gastrointestinal tract than cheese B, made with Lb. rhamnosus ; the latter did not belong to the predominant microflora of one out of the two B cheeses. Urea-PAGE electrophoresis results indicated that adjunct lactobacilli enhanced the degradation of both α _S -casein (α _S -CN) and β-casein (β-CN). In the fresh cheese, hydrolysis of α _S -CN was more rapid than β-CN, and the free amino acid content of B and C was higher than in A. Lipolysis products were also higher in B and C than in A as ripening progressed, and the organoleptic characteristics of these cheeses resulted in higher scores, in the order C > B > A. Thus, making Kefalotyri-like cheese from caprine milk with probiotic lactobacilli, particularly Lb. paracasei ssp. paracasei, as adjunct can be considered an effective way of producing a cheese with a large number of probiotic cells.     

Isolation of lactic acid bacteria from Lighvan cheese, a semihard cheese made from raw sheep milk in Iran

The lactic acid bacteria contributing to Lighvan cheese ripening during the different stages of production were investigated. Isolated strains from different culture media were identified phenotypically to species and subspecies level. In total, 413 strains were isolated from raw milk, 1-day-old cheese and fully ripened cheese. The most abundant species belonged to Enterococcus faecium (87 isolates), Lactococcus lactis ssp. lactis (68 isolates), Enterococcus faecalis (55 isolates) and Lactobacillus plantarum (48 isolates). E. faecium, Lc. lactis and Lb. plantarum were the predominantly isolated strains from ripened cheese. Therefore, they may contribute considerably to the aroma and flavour development of Lighvan cheese.     

Yield and sensory properties of cheese made with milk from Holstein or Montbéliarde cows milked twice or once daily

The aim of this study was to evaluate the milk properties and the yield and sensory properties of Cantal cheese made with milk from Holstein or Montbéliarde cows milked once or twice daily. Sixty-four grazing cows [32 Holstein (H) and 32 Montbéliarde (M) cows] in the declining phase of lactation (157 d in milk) were allocated to 1 of 2 equivalent groups milked once daily (ODM) or twice daily (TDM) for 7 wk. The full-fat raw milk collected during 24 h from the 4 groups of cows (M-TDM, M-ODM, H-TDM, and H-ODM) was pooled and processed into Cantal cheese 4 times during the last 4 wk of the experimental period. In all, 16 cheeses were made (2 milking frequencies × 2 breeds × 4 replicates) and analyzed after a ripening period of 15 and 28 wk. The results showed that for both breeds, the pooled milk content of fat, whey protein, casein, total protein, and phosphorus as well as rennet clotting time and curd firming time were significantly higher with ODM cows, whereas the casein-to-total protein ratio was lower, and lactose, urea, calcium, and free fatty acids contents of milk remained unchanged. The acidification and draining kinetics of the cheese as well as cheese yields and the chemical and rheological properties of the ripened cheese were not significantly modified by milking frequency. For both breeds, the cheeses derived from ODM cows had a slightly yellower coloration but the other sensory attributes, except for pepper odor, were not significantly affected by milking frequency, thereby demonstrating that ODM does not have an adverse effect on the sensory properties of Cantal cheese. Compared with that of Holstein cows, milk from Montbéliarde cows resulted in a higher cheese yield (+1.250 kg/100 kg of milk) and ripened cheeses with lower pH, dry matter, calcium, sodium chloride, and water-soluble nitrogen concentrations. These cheeses had also a less firm and more elastic texture, a more acidic taste, and a yogurt/whey aroma.