Studies on the extraction of milk-clotting enzyme after solid-state fermentation (SSF) of wheat bran by a local strain of Aspergillus oryzae LSI were done. The extraction of the enzyme was found to be depended on different parameters like nature of extractant, soaking time, temperature etc. From different inorganic and organic extractants, calcium chloride (0.05%) and glycerol (40%) were found to be the best solvents for leaching out milk-clotting enzyme. The optimum volume of calcium chloride was 5 ml/g biomass. An extraction time of 180 min. at 30°C and 100 rpm was sufficient to extract out nearly 28% of the enzyme (2666.7 U/g biomass). Most of the enzyme (about 98.71%) was recovered in four repeated extractions.
In this work, the effect of solid-substrate fermentation with Aspergillus oryzae, Rhizopus oryzae and Bacillus subtilis of soybean seeds on bioactive phenolic compounds was studied. Among the analysed sample extracts several phenolic compounds, hydroxybenzoics, hydroxycinnamics and fl avonoids, such as fl avonols, fl avanones, isofl avones were identifi ed by HPLC-DAD-ESI/MS. The results obtained indicate that fermentation process carried out in seeds inoculated with different microorganisms produced signifi cant changes in fl avonoids and phenolic acids contents. A signifi cant increase in the content of phenolic acids was observed in the samples fermented with the different microorganisms with respect to soybean without fermentation and fermented naturally. Fermentation process produced also important changes in fl avonoids compounds, with a signifi cant formation in isofl avone aglycone contents such as daidzein, glycitein and genistein as a consequence of glucosidase activity of microorganism in this process, showing signifi cant differences (p<0.05) with respect to control. Therefore, this process was shown to be a good way to increase the phenolic content of soybean, which could confer health-promoting effects.