A concentrated form of cell free extract (CFE) derived from attenuated Lactococcus lactis supsb. lactis 303 CFE was encapsulated in liposomes prepared from two different proliposome preparations (Prolipo Duo and Prolipo S) using microfluidization. Entrapment efficiencies of 19.7 % (Prolipo S) and 14.0 % (Prolipo Duo) were achieved and the preparations mixed in the ratio 4 (Prolipo Duo):1 (Prolipo S). Cheddar cheese trials were undertaken evaluating the performance of CFE entrapped in liposomes, empty liposomes and free CFE in comparison to a control cheese without any CFE or liposomes. Identical volumes of liposome and amounts of CFE were used in triplicate trials. The inclusion of liposomes did not adversely impact on cheese composition water activity, or microbiology. Entrapment of CFE in liposomes reduced loss of CFE to the whey. No significant differences were evident in proteolysis or expressed PepX activity during ripening in comparison to the cheeses containing free CFE, empty liposomes or the control, as the liposomes did not degrade during ripening. This result highlights the potential of liposomes to minimize losses of encapsulated enzymes into the whey during cheese production but also highlights the need to optimize the hydrophobicity, zeta potential, size and composition of the liposomes to maximize their use as vectors for enzyme addition in cheese to augment ripening.
References
[1]
Wilkinson, M.G.; Kilcawley, K.N. Mechanisms of incorporation and release of enzymes into cheese during ripening. Int. Dairy J. 2005, 15, 817–830, doi:10.1016/j.idairyj.2004.08.021.
[2]
El Soda, M.; Pandian, S. Recent developments in accelerated cheddar cheese ripening. J. Dairy Sci. 1991, 74, 2317–2335, doi:10.3168/jds.S0022-0302(91)78405-1.
[3]
Azarnia, S.; Robert, N.; Lee, B. Biotechnological methods to accelerate cheddar cheese ripening. Crit. Rev. Biotechnol. 2006, 26, 121–143, doi:10.1080/07388550600840525.
[4]
Kailasapathy, K.; Lam, S.H. Application of encapsulated enzymes to accelerate cheese ripening. Int. Dairy J. 2005, 15, 929–939, doi:10.1016/j.idairyj.2004.11.006.
[5]
Azarnia, S.; Lee, B.H.; St-Gelais, D.; Champagne, C.P.; Kilcawley, K.N. Effect of free or encapsulated recombinant aminopeptidase of Lactobacillus rhamnosus S93 on acceleration of cheddar cheese ripening. Food Biotechnol. 2010, 24, 135–149, doi:10.1080/08905431003784853.
[6]
Thompson, A.K.; Singh, H. Preparation of liposomes from milk fat globule membrane phospholipids using a microfluidizer. J. Dairy Sci. 2006, 89, 410–419, doi:10.3168/jds.S0022-0302(06)72105-1.
[7]
Lariviere, B.; Soda, M.E.; Soucy, Y.; Trepanier, G.; Paquin, P.; Vuillemard, J.C. Microfluidized liposomes for the acceleration of cheese ripening. Int. Dairy J. 1991, 1, 111–124, doi:10.1016/0958-6946(91)90003-Q.
Dufour, P.; Vuillemard, J.C.; Laloy, E.; Simard, R.E. Characterization of enzyme immobilization in liposomes prepared from proliposomes. J. Microencapsul. 1996, 13, 185–194, doi:10.3109/02652049609052906.
[10]
Kheadr, E.E.; Vuillemard, J.C.; El-Deeb, S.A. Acceleration of Cheddar cheese lipolysis by using liposome-entrapped lipases. J. Dairy Sci. 2002, 67, 485–492.
[11]
Kheadr, E.E.; Vuillemard, J.C.; El-Deeb, S.A. Impact of liposome-encapsulated enzyme cocktails on the cheddar cheese ripening. Food Res. Int. 2003, 36, 241–252, doi:10.1016/S0963-9969(02)00166-7.
[12]
Laloy, E.; Vuillemard, J.C.; Simard, R.E. Characterization of liposomes and their effect on the properties of Cheddar cheese during ripening. Lait 1998, 78, 401–412, doi:10.1051/lait:1998439.
[13]
Picon, A.; Serrano, C.; Gaya, P.; Medina, M.; Nunez, M. The effect of liposomes-encapsulated cryopsins on Manchego cheese ripening. J. Dairy Sci. 1996, 79, 1699–1705, doi:10.3168/jds.S0022-0302(96)76535-9.
[14]
Nongonierma, A.B.; Abrlova, M.; Fenelon, M.A.; Kilcawley, K.N. Evaluation of two food grade proliposomes to encapsulate an extract of a commercial enzyme preparation by microfluidization. J. Agric. Food Chem. 2009, 57, 3291–3297.
[15]
Jackson, L.S.; Lee, K. Microencapsulation and the food industry. Lebensm. Wiss. Technol. 1991, 24, 289–297.
[16]
Zeisig, R.; Cammerer, B. Single Core Encapsulation. Liposomes in Food Industry. In Microencapsulation of Food Ingredients, 1st; Vilstrup, P., Ed.; Leatherhead Publishing: Surrey, UK, 2001; pp. 101–119.
[17]
Kim, H.-H.Y.; Baianu, I.C. Novel liposome microencapsulation techniques for food applications. Trends Food Sci. Technol. 1991, 2, 55–61, doi:10.1016/0924-2244(91)90622-P.
[18]
Wilkinson, M.G. Acceleration of Cheese Ripening. In Cheese: Chemistry, Physics and Microbiology; Fox, P.F., Ed.; Chapman and Hall: London, UK, 1993; pp. 523–553.
[19]
Connor, J.; Yatvin, M.B.; Huang, L. pH-Sensitive liposomes: Acid-induced liposome fusion. Proc. Natl. Acad. Sci. 1984, 81, 1715–1718, doi:10.1073/pnas.81.6.1715.
[20]
Bainville, C.; Vuillemard, J.C.; Lacroix, C. Comparison of different methods for fortifying Cheddar cheese with vitamin D. Int. Dairy J. 2000, 10, 375–382, doi:10.1016/S0958-6946(00)00054-6.
[21]
Lawrence, R.C.; Gilles, J.; Creamer, L.K.; Crow, V.L.; Heap, H.A.; Honoré, C.G.; Johnston, K.A.; Samal, P.K. Cheddar Cheese and Related Dry-Salted Cheese Varieties. In Cheese: Chemistry, Physics and Microbiology, 3rd; Fox, P.F., McSweeny, P.L.H., Cogan, T.M., Guinee, T.P., Eds.; Elsevier Academic Press: London, UK, 2004; pp. 71–102.
[22]
O’Donovan, C.M.; Wilkinson, M.G.; Guinee, T.P.; Fox, P.F. An investigation of the autolytic properties of three lactococcal strains during cheese ripening. Int. Dairy J. 1996, 6, 1149–1165, doi:10.1016/S0958-6946(96)00024-6.
[23]
Crow, V.L.; Coolbear, T.; Gopal, P.K.; Martley, F.G.; McKay, L.L.; Riepe, H. The role of autolysis of lactic acid bacteria in the ripening of cheese. Int. Dairy J. 1995, 5, 855–875, doi:10.1016/0958-6946(95)00036-4.
[24]
McSweeney, P.L.H.; Sousa, M.J. Biochemical pathways for the production of flavour compounds in cheeses during ripening: A review. Lait 2000, 80, 293–324, doi:10.1051/lait:2000127.
[25]
Singh, T.K.; Drake, M.A.; Cadwallader, K.R. Flavor of Cheddar cheese: A chemical and sensory perspective. Compr. Rev. Food Sci. Food Saf. 2003, 2, 166–189, doi:10.1111/j.1541-4337.2003.tb00021.x.
[26]
Kilcawley, K.N.; Nongonierma, A.B.; Hannon, J.A.; Doolan, I.A.; Wilkinson, M.G. Evaluation of commercial enzyme systems to accelerate Cheddar cheese ripening. Int. Dairy J. 2012, 26, 50–57, doi:10.1016/j.idairyj.2012.03.015.
[27]
Drake, M.A.; McIngvale, S.C.; Gerard, P.D.; Cadwallader, K.R.; Civille, G.V. Development of a descriptive language for Cheddar cheese. J. Food Sci. 2001, 66, 1422–1427, doi:10.1111/j.1365-2621.2001.tb15225.x.
[28]
Young, N.D.; Drake, M.; Lopetcharat, K.; McDaniel, M.R. Preference mapping of cheddar cheese with varying maturity levels. J. Dairy Sci. 2004, 87, 11–19, doi:10.3168/jds.S0022-0302(04)73136-7.
[29]
Laridi, R.; Kheadr, E.E.; Benech, R.-O.; Vuillemard, J.C.; Lacroix, C.; Fliss, I. Liposome encapsulated nisin Z: Optimisation, stability and release during milk fermentation. Int. Dairy J. 2003, 13, 325–336, doi:10.1016/S0958-6946(02)00194-2.
[30]
Wilkinson, M.G.; Guinee, T.P.; O’Callaghan, D.M.; Fox, P.F. Autolysis and proteolysis in different strains of starter bacteria during Cheddar cheese ripening. J. Dairy Res. 1994, 61, 249–262, doi:10.1017/S0022029900028260.
[31]
Habibi-Najafi, M.B.; Lee, B.H. Proline-specific peptidases of Lactobacillus casei subspecies. J. Dairy Sci. 1994, 77, 385–392, doi:10.3168/jds.S0022-0302(94)76964-2.
[32]
Kilcawley, K.N.; Wilkinson, M.G.; Fox, P.F. Determination of key enzyme activities in commercial peptidase and lipase preparations from microbial or animal sources. Enzym. Microb. Technol. 2002, 31, 310–320, doi:10.1016/S0141-0229(02)00136-9.
[33]
Cogan, T.M.; O’Dowd, M.; Mellerick, D. Effects of pH and sugar on acetoin production from citrate by Leuconostoc lactis. Appl. Environ. Microbiol. 1981, 41, 1–8.
[34]
Hickey, D.K.; Kilcawley, K.N.; Beresford, T.P.; Sheehan, E.M.; Wilkinson, M.G. The influence of a seasonal milk supply on the biochemical and sensory properties of Cheddar cheese. Int. Dairy J. 2006, 16, 679–690, doi:10.1016/j.idairyj.2005.10.017.
[35]
Kuchroo, C.N.; Fox, P.F. Soluble nitrogen in cheese: Comparison of extraction procedures. Milchwiss. 1982, 37, 331–335.
[36]
Gee, V.L.; Carey, M.A.; Auty, M.A.E. A new cryo-STEM method for imaging food colloids in the scanning electron microscope. Scanning 2010, 32, 150–154, doi:10.1002/sca.20190.
Meilgaard, M.; Civille, G.; Carr, B. Descriptive Analysis Techniques. In Sensory Evaluation Techniques; CRC Press: New York, NY, USA, 1999; pp. 173–183.
[39]
Carunchiawhetstine, M.E.; Karagul-Yuceer, Y.; Avsar, Y.K.; Drake, M.A. Identification and quantification of character aroma components in fresh chevre-style goat cheese. J. Food Sci. 2003, 68, 2441–2447, doi:10.1111/j.1365-2621.2003.tb07043.x.
[40]
Drake, M.A.; Yates, M.D.; Gerard, P.D.; Delahunty, C.M.; Sheehan, E.M.; Turnbull, R.P.; Dodds, T.M. Comparison of differences between lexicons for descriptive analysis of Cheddar cheese flavour in Ireland, New Zealand, and the United States of America. Int. Dairy J. 2005, 15, 473–483, doi:10.1016/j.idairyj.2004.09.006.
[41]
Kang, E.J.; Smith, T.J.; Drake, M.A. Alternative bleaching methods for Cheddar cheese whey. J. Food Sci. 2012, 77, C818–C823, doi:10.1111/j.1750-3841.2012.02774.x.
