Novozyme 435 and Lipozyme RM IM Preferably Esterify Polyunsaturated Fatty Acids at the sn‐2 Position

Abstract

This work aims at studying the impact that the degree of unsaturation of fatty acids (FA) with the same chain length exerts on the esterification yield and the regiodistribution of FA in the glycerol backbone. To that end, triacylglycerols (TAG) are produced by esterifying several mixtures (stearic, oleic, and alpha-linolenic acids) employing Novozyme 435 and Lipozyme RM IM. All esterifications achieve a high content of TAG (>69 mol%) with Lipozyme RM IM being more active. Both lipases initially esterify stearic acid, while linolenic acid is preferably esterified in the last step of the esterification. Additionally, the higher the degree of unsaturation, the higher the concentration at the position sn-2. This trend is more marked for Novozyme 435. The structure of the FA causes steric hindrance resulting in competitive-like behavior among the FA. Hence, the selectivity of lipases might be affected by the composition of the substrate. Practical Applications: Omega-3 FA have a high nutritional value and are industrially employed to produce concentrates and structured or functional lipids. The regiodistribution of FA in the glycerol determines its bioavailability; those linked to the central position are more efficiently metabolized while those in the sn-1 and sn-3 positions are merely stored or excreted. Structured lipids are produced employing lipases, hence the regioselectivity of the lipases is a key point for the production. The results show that, for the same chain length, the regioselectivity of lipases can be affected by the composition of the substrate and that polyunsaturated FA tend to be esterified in the central position when employing Novozyme 435 and Lipozyme RM IM.

Progress and perspectives of enzymatic preparation of human milk fat substitutes

“…However, Novozym 435 exhibited preference for short chain fatty acids and long chain fatty acids, as reported by Wu et al [62]. Duan et al [63], Misbah et al [64] and Rivero-Pino et al [65] found that many lipases had substrate specificity toward triacylglycerols with short or medium carbon chain during the hydrolysis of triacylglycerols. While for the hydrolysis of fatty acid esters, lipases expressed different selectivity.…”

New Forms of Neuroactive Phospholipids for DHA Enrichment in Brain

“…In turn, for grapeseed oil, palmitic acid (16:0) was the predominant FA released [ 66 ]. Likewise, when the lipase from R. miehei was used to produce TAG enriched with PUFA throughout an esterification reaction, Rivero-Pino et al [ 78 ] reported that this enzyme initially esterifies stearic acid (18:0), while linolenic acid (18:3 n-3) is preferably esterified in the last step of the esterification, suggesting that it has a slight selectivity toward unsaturated FA.…”

Fabrication, characterization, and purification of nutraceutical diacylglycerol components from Camellia oil

“…Rivero‐Pino et al. (2020) studied the impact of several mixtures (stearic, oleic, and alpha‐linolenic acids) on the TAG production by esterifying with Novozym 435 and Lipozyme RM IM. The results showed that Lipozyme IM RM partially preferred to esterify unsaturated FA while N435 showed a random behavior in a solvent‐free system.…”