Dipeptide synthesis in near-anhydrous organic media: Long-term stability and reusability of immobilized Alcalase

作者：Vossenberg P^*; Beeftink H H; Nuijens T; Quaedflieg P J L M; Stuart M A Cohen; Tramper J

来源：Journal of Molecular Catalysis B: Enzymatic , 2013, 93: 23-27.

DOI：10.1016/j.molcatb.2013.03.014

• Immobilized protease
• Operational stability
• Reusability
• Molecular sieves
• Near-anhydrous
• Mechanical damage

The long-term stability and re-use of Alcalase covalently immobilized onto macroporous acrylic beads (Coy) in tetrahydrofuran (THF) were investigated. Coy can be used to synthesize dipeptides under near-anhydrous conditions in THF. Coy was incubated with and without molecular sieves (beads or powder) in THF, in order to investigate whether its stability is affected by the presence of molecular sieves. After different incubation periods, the enzyme activity was determined in an aqueous environment. In addition, Coy was repeatedly recycled to examine its reusability. Without molecular sieve beads, Coy hardly inactivated in THF. With molecular sieve beads, Coy lost activity over time. Incubated Coy samples were rotated on a blood rotator, entailing mechanical forces between Coy and the molecular sieve beads. Mechanical damage of Coy by the molecular sieve beads was found to be the main reason for the instability of Coy. During reuse, intermediate rehydration of Coy also caused a small but significant activity loss.

• 出版日期2013-9