Production of Human Milk Fat Substitutes by Interesterification of Tripalmitin with Ethyl Oleate Catalyzed by Candida parapsilosis Lipase/Acyltransferase

Abstract

In human milk fat, the saturated fatty acids, namely palmitic acid, are located at the sn-2 position of triacylglycerols (TAG) while unsaturated fatty acids (e.g. oleic acid) are esterified at position sn-1,3. Thus, sn-1,3-dioleoyl-2-palmitoylglycerol (OPO) is the target TAG to be used as human milk fat substitutes (HMFS) in infant formulas. In this study, the noncommercial recombinant lipase/acyltransferase from Candida parapsilosis (CpLIP2) was immobilized in Accurel MP1000, and used as a biocatalyst for the interesterification of tripalmitin with ethyl oleate in a solvent-free medium, to obtain structured lipids used as HMFS. Different molar ratios (MR) of ethyl oleate to tripalmitin (2:1-8:1) were used. After 4 h reaction at 60 C, about 30 mol% of oleic acid incorporation was already observed for all tested MR. An apparent equilibrium was reached after 8-24 h, with 32-51 mol% final incorporation, increasing with the MR. The incorporation of oleic acid into TAG was compared with the maximum predicted values when a random or a sn-1,3-regioselective biocatalyst was used. The obtained values are consistent with the maximum incorporation expected for a sn-1,-3-regioselective enzyme. In fact, the amount of oleic acid at position sn-2 was approximately 15% for all the MR tested, which is explained by the acyl migration phenomenon. CpLIP2 exhibited higher activity than most commercial immobilized lipases (e.g. faster reaction in solvent-free media, low enzyme load, and low MR needed), and showed a recognized sn-1,3 regioselective behavior. Keywords Candida parapsilosis lipase acyltransferase Á Ethyl oleate Á Human milk fat substitute Á sn-1,3 regioselective enzyme Á Structured lipids J Am Oil Chem Soc (2019) 96: 777-787. Abbreviations CpLIP2 recombinant lipase/acyltransferase from Candida parapsilosis FAME fatty acid methyl ester HMF human-milk fat HMFS human-milk fat substitutes MR molar ratio(s) omega-3 PUFA omega-3 polyunsaturated fatty acid(s) OPO 1,3-dioleoyl-2-palmitoylglycerol SL structured lipid(s) TAG triacylglycerol(s)

Produção de sucedâneo da gordura do leite humano por interesterificação enzimática: uma revisão

“…Dos nove trabalhos específicos encontrados, seis apresentaram como objetivo a produção de um substituto da gordura do leite humano (HFMS) por processo de interesterificação enzimática a partir de diversas fontes de óleos e gorduras naturais (Yuan et al, 2020;Tecelão et al, 2019;Ghosh et al, 2016;Zou et al, 2016;Djajasoepena et al, 2015;Lee et al, 2015). Yuan et al, (2020), por exemplo sintetizaram Lípidos estruturados enriquecidos com TAGs de cadeia média e longa (MLCTs) por interesterificação de óleo de côco e de Bagre, usando a enzima NS 40086.…”

“…A mistura de PSF e FO na razão molar de 2: 1 com teor de PA de 69,70% e 12 h de reação produziu o produto desejado com 75,98% de PA em sn-Posição 2, 0,27% de AA, 3,43% de EPA e 4,25% de DHA e com ponto de fusão de 42 ° C. Este estudo retratou uma preparação bem-sucedida de TAG contendo composição única de FA, ou seja, ≥ 70% do PA, por peso, foram esterificados na posição sn-2, que poderia ser usada em formulação infantil com benefícios para a saúde de n-3 PUFAs. Tecelão et al, (2019), utilizaram a lipase / aciltransferase de Candida parapsilosis (CpLIP2) não comercial imobilizada em Accurel MP1000, como biocatalisador para a interesterificação de tripalmitina com oleato de etila em meio livre de solvente, para obtenção de lipídios estruturados usado como HMFS. A quantidade de ácido oleico na posição sn-2 foi de aproximadamente 15% para todas as razões molares testadas.…”

Enzymatic Synthesis of Human Milk Fat Substitute - A Review on Technological Approaches

“…Nevertheless, this issue is specific, depending on the type and proportion of oil ( 5 , 9 ). Table 4 ( 43 , 55 - 59 , 66 , 69 , 86 - 92 ) summarizes the reaction conditions for HMFS synthesis by transesterification.…”

“…42,[54][55][56][57][58]65,68,[85][86][87][88][89][90][91] summarizes the reacting conditions of HMFS synthesis by transesterification.…”

Synthesis of Dietetic Structured Lipids from Spent Coffee Grounds Crude Oil Catalyzed by Commercial Immobilized Lipases and Immobilized Rhizopus oryzae Lipase on Biochar and Hybrid Support

“…Low-calorie SL can be produced enzymatically either (i) by acidolysis of a TAG or oil, containing long-chain fatty acids, with a medium-chain fatty acid [26], or (ii) by interesterification of a TAG or oil with methyl or ethyl esters (fatty acid methyl esters, FAME, or fatty acid ethyl esters, FAEE) of medium-chain fatty acids [27,28] or medium-chain TAG [29]. Reactions catalyzed by enzymes are carried out under mild temperatures, reducing energy consumption, leading to a decrease in the loss of temperature-sensitive molecules (substrates and/or products), and resulting in more environment-friendly and safer processes.…”