Enzyme Engineering for High‐Yielding Amide Formation: Lipase‐Catalyzed Synthesis of N‐Acyl Glycines in Aqueous Media

Abstract

Amide syntheses remain a key challenging green chemistry reaction. For instance, green synthesis of N-acyl glycines as biosurfactants and therapeutics is highly desirable to replace chemical pathways using toxic phosgene. Herein, we report a novel concept for enzymatic amidation in an aqueous system via glycerol activation of fatty acids and theirsubsequent aminolysis with glycine to synthesize N-acyl glycines. We then engineer an enzyme (proRML) by reshaping its catalytic pocket to enhance its aminolysis activity and catalytic efficiency by 103-fold and 465-fold, respectively. The evolved proRML (D156S/L258K/L267N/S83D/L58K/ R86K/W88V) catalyzed the amidation of a fatty acid with glycine to give N-lauroylglycine with high yield (80 %). It accepts a broad range of medium-to longchain fatty acids (C 8 -C 18 ), giving high yields of Ndecanoyl-, N-myristoyl-, and N-oleoylglycine. The developed amidation concept may be general, and the engineered enzyme is useful for the green synthesis of N-acyl glycines.

Amorphous Fe0.7Cu0.3Ox as a Bifunctional Catalyst for Selective Hydration of Nitriles to Amides in Water

“…Among the various methods − for amide synthesis, nitrile hydration is recognized as one of the most atom-efficient approaches. , Design of novel catalysts for nitrile hydration has attracted considerable attention; homogeneous and heterogeneous catalysts of noble metals have been reported, such as palladium, − ruthenium, − rhodium, , gold, and silver . Among them, several efficient homogeneous catalysts reported in nitrile hydration are designed with the “bifunctional catalysis” concept.…”

Immobilization of Lipases Using Poly(vinyl) Alcohol

“…Within the group of hydrolases are lipases, whose natural function is the hydrolysis of triglycerides [2,3]. From a practical point of view, lipases (triacylglycerol hydrolases, EC 3.1.1.3) are important biocatalysts in several applications, such as the synthesis of chiral drug intermediates [4] and nutraceutical lipids [5], bioconversion of oils and fats [6], and production of biodegradable polymers based on the ability of lipases for esterification, transesterification [7,8], aminolysis [9], and hydrolysis reactions [10]. The sources of lipases are found mainly in animals, plants, and microorganisms, where they play an essential role in their metabolism [11][12][13].…”