Notes

1. The solution may become cloudy at temperatures below 20°C but may restored by warming at 30°C and mixing.

2. Before mincing the beef/pork muscle, remove the fat and connective tissue in aseptic conditions, cut the lean muscle into small cubes (3 cm), and keep at -30°C until use.

3. To ensure the development of lactic acid bacteria in this system, it is necessary to add a carbon source.

4. This experimental system contain approx 4.5 g/L of protein.

5. The concentration of this growth factor should be determined for each lactic acid bacterium.

6. To avoid interference with the sarcoplasmic system, an optical density of 680 nm was selected.

7. After the TCA treatment, samples can be frozen at -20°C for further use.

References

1. Aristoy, M. C. and Toldra F. (1991) Deproteinization techniques for HPLC amino acid analysis in fresh pork muscle and dry cured ham. J. Agric. Food Chem. 39, 1792-1795.

2 De Masi, T. M., Wardlaw, F. B., Dick, R. L., and Ancton, J. C. (1990) Non protein nitrogen (NPN) and free amino acid contents of dry fermented sausages. Meat Sci. 27, 1-12.

3 Fadda, S., Sanz, Y., Vignolo, G., Aristoy, M-C., Oliver, G., and Toldra, F. (1999) Characterization of muscle sarcoplasmic and myofibrillar protein hydrolysis caused by Lactobacillus plantarum. Appl. Environ Microbiol. 65, 3540-3546.

4 Hammes, W. P., Bantleon, A., and Min, S. (1990) Lactic acid bacteria in meat fermentation. FEMS Microbiol. Rev. 87, 165-174.

5 Johansson, G., Berdague, J. L., Larsson, M., Tran, N., and Borch, E. (1994) Lipolysis, proteolysis and formation of volatile components during ripening of a fermented sausage with Pediococcus pentosaceus and Staphylococcus xylosus as starter cultures. Meat Sci. 38, 203-218.

6 Montel, M. C., Seronine, M. P., Talon, R., and Hebraud, M. (1995) Purification and characterization of a dipeptidase from Lactobacillus sake. Appl. Environ. Microbiol. 61, 837-839.

7 Sanz, Y., Fadda, S., Vignolo, G., Aristoy, M-C., Oliver, G., and Toldra, F. (1999) Hydro-lytic action of Lactobacillus casei CRL705 on pork muscle sarcoplasmic and myofibrillar proteins. J. Agric. Food Chem. 47, 3441-3448.

8. Church, F. C., Swaisgood, H. E., Porter, H. D., and Castignani, G. L. (1983) Spectropho-tometric assay using o-phaldialdehyde for determination of proteolysis in milk and isolated milk proteins. J. Dairy Sci. 66, 1219-1227.

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