2.8 Substrate scope investigation
To demonstrate the substrate range of CRL@OSMD, the esterification of pine sterol with oleic, linoleic, linolenic acid, and mixed acid (oleic, linoleic, and linolenic acids in equimolar ratios) was studied under optimized reaction conditions.
2.9 Process scale-up and reusability studies
To explore the scalability of the immobilized CRL-catalyzed pine sterol ester synthesis process, the feed volume was increased up to 10 times under optimized reaction conditions. The synthesis of pine sterol easters was investigated for reusability under optimized conditions. At the end of the reaction, the immobilized enzyme was filtered, separated from the reaction system, and washed with 5 mL hexane 3 times. After drying, the resulting immobilized CRL was used directly in the next batch.