IMMOBILIZATION OF HYDROPEROXIDE LYASE FROM CHLORELLA

Hydroperoxide lyase (HPLS) isolated from microalgae is an enzyme that oxidatively cleaves 13-hydroperoxy-cis-9-trans-11-octadecadienoic acid to a C-13 oxocarboxylic acid and a small C-5 fragment, Acetone powder extracts from Chlorella pyrenoidosa and C, fusca that contained HPLS were partly purified by chromatography on DEAE-Sepharose CL-6B, Five c ommercially available gels were evaluated for their ability to immobil ize the HPLS preparations by determining their capacity for protein bi nding and the activity and stability of immobilized HPLS, It was found that Reacti-Gel (6X) and Affi-Gels 10, 15, 102 and 501 could bind 60- 90% of the available protein, However, HPLS activity was detected only when the enzyme was immobilized on Affi-Gels 10, 15 and 501, The stab ility of immobilized HPLS during storage at 5 degrees C for several mo nths was determined, Product yields with repeated use of the immobiliz ed preparations were also determined, These measurements demonstrated that Affi-Gel 10 and 501 are the best gels for the immobilization of H PLS, A pH study of HPLS immobilized on Affi-Gel 501 showed that enzymi c activity was retained from pH 6 to, pH 9, with maximal activity at p H 6.5.