One-factor-at-a-time optimisation of the aqueous extraction of the peroxidase from fresh Amazonian cacao beans
2020
Konagano, E. M. H. | da Costa, H. S. C. | de Souza, J. N. S. | Herman, C. A. N. P.
Peroxidase plays an important role in some enzymatic phenomena occurring during the fermentation and drying of cacao beans. Peroxidase reduces the bitterness, astringency, and acidity, and helps develop the characteristic colour and chocolate flavour of cacao beans. The present work thus aimed to optimise five parameters for the aqueous extraction of the peroxidase from fresh Amazonian cacao beans using the one-factor-at-a-time methodology. The following parameters were studied: pH (4 to 10), molarity (50 to 400 mM) of the extraction buffer solution, grinding time (10 to 120 s), homogenisation time after grinding (0 to 90 min), solid-to-liquid ratio between the cacao bean mass, and the extraction buffer solution volume (1:2 to 1:16). The results showed that the recommended pH and molarity of the extraction buffer solution to be 9 and at least 200 mM, respectively. The results also showed that the grinding time to be at least 90 s, and the optimal homogenisation time to be 60 min. The optimal solid-to-liquid ratio must be 1:8. Under these conditions, the activity of the peroxidase increased from 0.01 variation of the absorbance unit per minute (ΔUA/min) to approximately 0.15 ΔUA/min.
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