Investigation of methods of extraction, concentration and conservation of rumen content fibrolytic enzymes

This study was carried out to determine the potential of rumen microorganisms in samples taken from slaughterhouse to produce fibrolytic enzymes and to compare different methods for extraction, concentration and conservation of them. According to the results, the average specific activity of cellulase and xylanase were 7.5 and 16.5 U/mg protein respectively. The application of homogenizing and sonication methods to extract rumen content enzymes showed that the former had better performance in the liberation of enzymes. To determine optimum time length for the maximum release of enzymes from solid phases, various time lengths including one, 1.5 and two min were examined. Based on this experiment, one minute appeared the best. To attain a high level of concentration of enzymes obtained from rumen liquor, various procedures were applied. The results indicate that freeze drying and precipitation of enzyme using ammonium sulphate were the best methods, while trichloroacetic acid precipitation turned out to be the most inappropriate method due to improper effect on enzymatic activity. In order to examine the effect of temperature and time length on enzymes activities, some methods including liquid nitrogen, freezing in -70° and freezing in -20° were applied. The comparison of different methods of enzymes conservation indicated that applying liquid nitrogen for long term as well as keeping in the freezer (-70°) for medium term purposes were the best.