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Immobilization of hydroperoxide lyase from Chlorella
Author(s) -
Nuñez Alberto,
Armand Gary St.,
Foglia Thomas A.,
Piazza George J.
Publication year - 1997
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1997.tb00416.x
Subject(s) - chlorella pyrenoidosa , chemistry , acetone , chromatography , enzyme , chlorella , immobilized enzyme , sepharose , biochemistry , specific activity , enzyme assay , algae , biology , botany
Hydroperoxide lyase (HPLS) isolated from microalgae is an enzyme that oxidatively cleaves 13‐hydro‐peroxy‐ 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 commercially available gels were evaluated for their ability to immobilize the HPLS preparations by determining their capacity for protein binding 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 60Ő90% of the available protein. However, HPLS activity was detected only when the enzyme was immobilized on Affi‐Gels 10, 15 and 501. The stability of immobilized HPLS during storage at 5° C for several months was determined. Product yields with repeated use of the immobilized preparations were also determined. These measurements demonstrated that Affi‐Gel 10 and 501 are the best gels for the immobilization of HPLS. A pH study of HPLS immobilized on Affi‐Gel 501 showed that enzymic activity was retained from pH 6 to pH 9, with maximal activity at pH 6.5.