Purification and Properties of Protease from Brazilian Pine (Araucaria angustifolia (Bert) O. Ktze) Nuts

Abstract

Abstract: An orthogonal array design was used to optimize conditions for the extraction of protease from Brazilian pine nuts. Boric acid-borate buffer was found to be the best solvent for the extraction of protease, and its optimum pH was 9.0. The optimum extraction time and solid-to-solvent ratio were 60 min and 1:8 (m/V), respectively. Pure protease was obtained from crude extract using salting out with ammonium sulfate followed by DEAE-Sepharose FF column chromatography. After purification, the specific activity of protease increased 8.61-fold and the activity recovery was 21.65%. As shown by SDS-PAGE, the molecular mass of the enzyme was 33 kD. Enzymatic characterization demonstrated that it was an alkaline and its optimum reaction pH and temperature were 9.0 and 50 ℃, respectively. In addition, Mn2+ had a strong activating effect on the enzyme, whereas Ca2+, Mg2+ and Cu2+ could inhibit its activity.

Key words: Brazilian pine nut, protease, purification, protease activity

CLC Number:

References

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