Application of bio-based solvents for biocatalysed synthesis of amides with Pseudomonas stutzeri lipase (PSL)

Abstract

Bio-based solvents were investigated for the biocatalysed amidation reactions of various ester-amine combinations by Pseudomonas stutzeri lipase (PSL). Reactions were undertaken in a range of green and potentially bio-based solvents including terpinolene, p-cymene, limonene, 2-methyl THF, ɣ-valerolactone, propylene carbonate, dimethyl isosorbide, glycerol triacetate and water. Solvent screenings demonstrated the importance and potential of using non-polar bio-based solvents for favouring aminolysis over hydrolysis; whilst substrate screenings highlighted the unfavourable impact of reactants bearing bulky para- or 4-substituents. Renewable terpene-based solvents (terpinolene, p-cymene, D-limonene) were demonstrated to be suitable bio-based media for PSL amidation reactions. Such solvents could provide a greener and more sustainable alternative to traditional petrochemical derived non-polar solvents. Importantly, once the enzyme (either PSL or CALB) binds with a bulky para-substituted substrate, only small reagents are able to access the active site. This therefore limits the possibility for aminolysis to take place, thereby promoting the hydrolysis. This mechanism of binding supports the widely accepted ‘Ping Pong – Bi Bi’ mechanism used to describe enzyme kinetics. The work highlights the need to further investigate enzyme activity in relation to para- or 4-substituted substrates. A priority in PSL chemistry remains a methodology to tackle the competing hydrolysis reaction.

Green solvents for the formation of amide linkages

“…The same solvent showed great suitability for the biocatalyzed synthesis of amides, in combination with the enzymatic catalyst Pseudomonas stutzeri lipase (PSL). 69 The developed method was able to afford acceptable conversion.…”