Scale-up studies on immobilization of lactoperoxidase using milk whey for producing antimicrobial agent

Hypothiocyanite (oscn-),produced by lactoperoxidase (lpo) in the presence of scn- and h2o2,inhibits the growth of bacteria. this inhibition is called by lpo system (lpos). our laboratory scale study in previous experiment showed that whey immobilized on sp-sepharose fast flow (sp-ff) could produce oscn- continuously. then,the purpose of this study is to scale up continuous production of oscn- using immobilized whey. immobilized whey was generated by circulating various amounts of whey through sp-ff. to generate oscn-,10 ml of the substrate solution containing 0.5 mm scn- and 0.5 mm h2o2,was circulated through immobilized whey and followed by washing with pure water. the next cycle was done by circulating a fresh 10 ml of substrate solution at the same concentration. the result indicated that a stable immobilization efficiency of more than 90% was achieved in the sp-ff circulated with 300 ml or less of whey per gram of sp-ff. when stored at 4°c,immobilized whey retained 80% lpo activity until 3 weeks storage. the reaction solution discharged from immobilized whey was observed to contain approximately 0.4 mm oscn-. the experiment using 1.0 g of immobilized whey produced a stable 0.4 mm oscn- production and antimicrobial activity for at least 6 cycles. the increase in resin volume accompanied by the increase in whey volume resulted the extension of a stable oscn- production. the experiment using recycled sp-ff did not affect to the stability of oscn- production and antimicrobial activity. these results may open the way for the large-scale production of oscn-. © 2010 diponegoro university. all rights reserved.