Accelerated Cheddar cheese ripening with encapsulated proteinases

This work was undertaken to study the impact of bacterial and fungal proteinases encapsulated in liposomes on proteolysis and organoleptic properties of Cheddar cheese during ripening. Bacterial and fungal proteinases were encapsulated in liposomes with trapping efficiencies of 31.9 and 33.0%, respectively. The encapsulated bacterial proteinase was incorporated into Cheddar-cheese milk in order to reach enzyme concentrations of 5 x 10 -6 (B 1 ). 1 x 10 -5 (B 2 ) and 2 x 10 -5 (B 3 ) AU g -1 cheese curd, while encapsulated fungal proteinase was added in order to reach enzyme concentrations of 1 x 10 -6 (F 1 ), 2 x 10 -6 (F 2 ) and 4 x 10 -6 (F 3 ) AU g -1 cheese curd. Experimental cheeses showed slightly higher moisture content but lower protein content when compared to control cheeses. Rheological measurements revealed that experimental cheeses were slightly less firm but more brittle as compared to control cheeses. Proteolysis and texture development of experimental cheeses were faster than control cheeses. As observed by transmission-electron microscopy, liposome-treated cheeses exhibited less compact microstructure and liposomes were essentially located at the fat-casein interface. The elution patterns obtained from size-exclusion chromatography revealed that the accumulation of bitter and astringent peptides was higher in experimental cheeses than in control cheeses depending on the type and concentration of added enzyme. After 3 months ripening, the organoleptic properties of cheeses were markedly improved and no bitter off-flavour was detected among treatments except cheeses subjected to treatments B 3 and F 3 . The present study showed that micro-encapsulated enzymes can be successfully used to accelerate cheese ripening and to avoid drawbacks resulting from the use of free enzymes. It was also concluded that the addition of encapsulated bacterial proteinase (up to 1 x 10 -5 AU g -1 cheese curd) or encapsulated fungal proteinase (up to 2 x 10 -6 AU g -1 cheese curd) could be recommended to accelerate the flavour and texture development of Cheddar cheese.

[1]  L. Vassal,et al.  Utilisation de Rulactine en tant qu'agent d'affinage dans des fromages à pâte pressée , 1987 .

[2]  J. Vuillemard,et al.  Influence of the fat content of Cheddar cheese on retention and localization of starters , 1996 .

[3]  M. Desmazeaud,et al.  Etude du rôle des micro-organismes et des enzymes au cours de la maturation des fromages. II. - Influence de la présure commerciale , 1975 .

[4]  B. Law,et al.  Accelerated cheese ripening with food grade proteinases , 1982, Journal of Dairy Research.

[5]  R. C. Lawrence,et al.  Texture Development During Cheese Ripening , 1987 .

[6]  C. H. Amundson,et al.  Accelerated ripening of Gouda cheese made from ultrafiltered milk using a liposome entrapped enzyme and freeze shocked lactobacilli , 1989 .

[7]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[8]  S. Pandian,et al.  Recent developments in accelerated cheese ripening , 1991 .

[9]  M. E. Soda.  Accelerated maturation of cheese , 1993 .

[10]  J. Vuillemard,et al.  Characterization of enzyme immobilization in liposomes prepared from proliposomes. , 1996, Journal of microencapsulation.

[11]  M. Vijayalakshmi,et al.  High performance size exclusion liquid chromatography of small molecular weight peptides from protein hydrolysates using methanol as a mobile phase additive , 1986 .

[12]  V. Bottazzi,et al.  Proteolysis in cheese made with liposome-entrapped proteolytic enzymes , 1993 .

[13]  A. Picon,et al.  The Effect of Liposome Encapsulation of Chymosin Derived by Fermentation on Manchego Cheese Ripening , 1994 .

[14]  D. Stanley,et al.  HPLC Separation of Bitter Peptides from Cheddar Cheese , 1982 .

[15]  P. Fox,et al.  Contribution of plasmin to Cheddar cheese ripening: effect of added plasmin , 1992, Journal of Dairy Research.

[16]  J. Gripon,et al.  Liposomes as Proteinase Carriers for the Accelerated Ripening of Saint‐Paulin Type Cheese , 1988 .

[17]  S. Visser.  Proteolytic Enzymes and Their Relation to Cheese Ripening and Flavor: An Overview , 1993 .

[18]  D. McMahon,et al.  Microstructure of Mozzarella Cheese During Manufacture , 1993 .

[19]  G. Jong.  The Influence of Dispersal Pattern On the Evolution of Fecundity , 1981 .

[20]  N. Olson,et al.  Accelerated ripening of Gouda cheese. 2. Effect of freeze-shocked Lactobacillus helveticus on proteolysis and flavor development. , 1987 .

[21]  Philip Smith,et al.  Rheology and microstructure of low-fat Mozzarella cheese , 1993 .

[22]  G. Puglisi,et al.  Neutrase entrapment in stable multilamellar and large unilamellar vesicles for the acceleration of cheese ripening. , 1995, Journal of microencapsulation.

[23]  L. D. Jong.  Protein breakdown in soft cheese and its relation to consistency. 3. The micellar structure of Meshanger cheese , 1978 .

[24]  P. Fox,et al.  Soluble nitrogen in Cheddar cheese: comparison of extraction procedures , 1982 .

[25]  K. Morihara.  Comparative specificity of microbial proteinases. , 1974, Advances in enzymology and related areas of molecular biology.

[26]  D. Stanley,et al.  Cheddar Cheese Made With Bovine Pepsin II. Texture—Microstructure—Composition Relationships , 1977 .

[27]  Mi Sun Kim,et al.  Citrate Inhibition of Aminopeptidase in Commercial Fungal Protease Preparations Used to Accelerate Cheese Ripening , 1989 .

[28]  G. Milliken,et al.  Effects of Commercial Food Grade Enzymes on Proteolysis and Textural Changes in Granular Cheddar Cheese , 1987 .

[29]  J. A. Elliott,et al.  Isolation of Bitter and Astringent Fractions from Cheddar Cheese , 1971 .

[30]  C. Serrano,et al.  The effect of liposome-encapsulated cyprosins on manchego cheese ripening , 1996 .

[31]  Alice McCarthy,et al.  Proteolytic and rheological changes during ageing of cheese analogues made from rennet caseins , 1994 .

[32]  Roger K. Abrahamsen,et al.  Influence of liposome-encapsulated Neutrase and heat-treated lactobacilli on the quality of low-fat Gouda-type cheese , 1995, Journal of Dairy Research.

[33]  L. D. Jong.  The influence of the moisture content on the consistency and protein breakdown of cheese , 1978 .

[34]  J. Vuillemard,et al.  Characterization of liposomes and their effect on the properties of Cheddar cheese during ripening , 1998 .

[35]  B. Law,et al.  Use of liposomes for proteinase addition to Cheddar cheese , 1985, Journal of Dairy Research.

[36]  R. Puchades,et al.  Size-Exclusion HPLC Separation of Bitter and Astringent Fractions from Cheddar Cheese Made with Added Lactobacillus Strains to Accelerate Ripening , 1989 .

[37]  M. Tunick,et al.  Cheddar and cheshire cheese rheology , 1990 .

[38]  P. Paquin,et al.  Microfluidized liposomes for the acceleration of cheese ripening , 1991 .

[39]  M. L. Anson,et al.  THE ESTIMATION OF PEPSIN, TRYPSIN, PAPAIN, AND CATHEPSIN WITH HEMOGLOBIN , 1938, The Journal of general physiology.