Immobilization of enantioselective lipase on soluble supports for kinetic resolution of drug intermediates

Abstract

The microbial lipase, Arthrobacter sp. lipase (MTCC 5125), from the Indian Institute of Integrative Medicine repository, is known as an effective catalyst for high enantioselective kinetic resolution of drug intermediates. The ABL was immobilized on water-soluble linear supports by covalently binding it to the epoxy groups on the N-vinyl pyrrolidone/allyl glycidyl ether and N-vinyl pyrrolidone/glycidyl methacrylate copolymers. The immobilized lipase, on different soluble supports, had 90–110 mg/g protein binding and 500–700 U/g hydrolysis activities for tributyrin substrate. These copolymers had soluble/insoluble characteristics in different pH ranges, which is an advantage over insoluble copolymers. A soluble polymer at neutral pH provided better accessibility to the immobilized enzyme, which was recovered by precipitation at pH 2–3 for reuse. Kinetic resolution of racemic acyl derivatives of chiral auxiliaries and drug intermediates, namely, phenyl ethanol, aminoalcohol, and fluoxetine intermediate resulted in a significant enhancement in enantioselectivity (99%).

A hydrolase with esterase activity expressed from a fosmid gene bank prepared from DNA of a North West Himalayan glacier frozen soil sample

“…3c). Lipases/esterase showing enantioselectivity have also been reported earlier (Chaubey et al 2012;Rasool et al 2005).…”

“…Enantiospecificity of ME-003 for the resolution of racemic drug intermediates/precursors (1-phenyl ethanol, flouxetine, and ethambutol) was analysed by following the protocol reported earlier (Chaubey et al 2012).…”

“…Previously, the sequence-based metagenomics, e.g., next-generation sequencing (NGS) strategies have been extremely successful in identification of novel enzymes, i.e., nitrilases, cellulases, carboxylesterases, lipases and other industrially important enzymes (Beloqui et al 2006;Liebl et al 2014;Lorenz and Eck 2005;Robertson et al 2004;Steele et al 2009). Esterases (EC.3.1.1.1, carboxyl ester hydrolases) are hydrolytic enzymes that catalyse the hydrolysis of various types of exogenous and endogenous esters, preferably esters composed of short chain of fatty acids into acid and alcohol (Bhavith et al 2014;Bornscheuer 2002). These industrially important enzymes have major physiological importance being extensively distributed in ecosystem, for example, in bacteria, archaea, and eukaryotes, and their industrial applications include synthesis of novel esters, resolution of chiral carboxylic acids, primary, secondary and tertiary alcohols, resolution of chiral drugs, etc.…”

“…(Margolin 1993;Montella et al 2012;Sayer et al 2015). These enzymes have broad application in the industry, based on the grounds of their versatility, adaptability, stereo-selectivity and their ability to improve synthetic reactions in organic solvents (Bornscheuer 2002). The first ever experimental structure of an esterase enzyme belongs to acetyl-choline esterase (Sussman et al 1991).…”

A review on Arthrobacter sp. lipase: A versatile biocatalyst for the kinetic resolution to access enantiomerically pure/enriched compounds

“…However, immobilization, by covalent bonding through glutaraldehyde, on magnetic and nonmagnetic sol‐gel composite supports provided 50–70 U/g activity and 28–30 mg/g protein binding. Further studies using N ‐vinyl pyrrolidone‐allylglycidyl ether (ANP)/ N ‐vinyl pyrrolidone‐glycidyl methacrylate (GNP) copolymers revealed that immobilization of ABL on soluble support ANP1 and GNP1 having higher epoxy content provided more binding capacity (100–110 mg/g support) and activity (700 U/g in ANP1 and 600 U/g in ANP2) as compared to ANP2 and GNP2 type supports (protein binding of 90–95 mg/g support and activity 600 U/g in ANP2 and 500 U/g in GNP2) 35 …”