Hydroperoxide lyase (HPL) from amaranth tricolor was immobilized on the ceramic hydroxyapatite (CHT). A maximum activity of 2.85�0.1 U/g CHT (wet) and a yield of 80% were obtained under optimized HPL-CHT coupling condition. The optimum pH was 6.0, 6.0 and 7.5 for free, membrane and CHT-immobilized HPL while the optimum reaction temperature was 30, 30 and 35oC, respectively. After incubating at 55oC for 30 min, the residual activity of CHT-immobilized HPL was 64.13%, while those of free and membrane HPL were 0. Km values showed CHT (239.10 �M) imposed a higher substrate diffusion resistance than membrane (171.57 �M). Furthermore, recycling assays showed that CHT-immobilised HPL was more stable than membrane HPL. When the substrate concentration varied in the range of 66.7-400 �M, CHT-immobilised HPL retained above 90% of the initial activity after five reaction cycles while the activity of membrane bound HPL was decreased to zero after three reaction cycles.

Hydroperoxide lyase immobilized on ceramic hydroxyapatite with better stability than the membrane bound hydroperoxide lyase

244th National Meeting (2012)

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