Achieving cinnamic acid amides in water by a variant of acyltransferase from Mycobacterium smegmatis and its immobilized form using Ni-NTA modified aspen powder as a carrier

Abstract

An aqueous N-acylation reaction for preparing cinnamic acid amides was realized by using a variant of acyltransferase from Mycobacterium smegmatis (MsAcT-L12A), whereas the wild-type MsAcT showed no activity. MsAcT-L12A exhibited broad substrate adaptability, and preferred the substrates with electron-donating group. When the vinyl cinnamate (1a, 40 mM) and p-methoxyaniline (2a, 4 mM) were involved in the reaction, the excellent yield reached to 86.7 % ± 2.1 % within 3 h by MsAcT-L12A (1 mg_pro./mL) in a PBS buffer (100 mM, pH 8.0) at 25 °C. The aqueous N-acylation reaction could be further improved by using an immobilized MsAcT-L12A. The biomass aspen powder (AP) as a carrier provided a low-cost, green, and environmental-friendly immobilization strategy. After it was modified by Ni-NTA, the obtained Ni-NAP could realize one-step purification and immobilization of MsAcT-L12A. The accomplished MsAcT-L12A-Ni-NAP exhibited excellent stability and recyclability, and retained its relative yield as 83.3 % ± 2.2 % even after the 7th cycle of reuse. Using only PBS buffer as a reaction medium, the operation for MsAcT-L12A-catalyzed acyl transfer was greatly simplified, and the improved stabilities of MsAcT-L12A-Ni-NAP could enhance its application potential.

Keywords: Acyltransferase from Mycobacterium smegmatis; Aqueous N-acylation reaction; Cinnamic acid amides.