At the end of incubation, the culturemedium
was filtered through the cotton cloth to remove soybean, whichwas
followed by centrifugation at 5600×g, 4◦C for 20 min and filtering
through a syringe filter (0.45m) to remove the bacteria. The liq
liquid
broth of the crude enzyme was used to assay for milk-clotting
activity. To investigate the factors affecting MCE production, the
cultivationwas carried out in themedium and conditions indicated
above except that the factors were varied by one-factor-at-a-time
fashion.