Production of Human Milk Fat Substitutes by Lipase-Catalyzed Acidolysis: Immobilization, Synthesis, Molecular Docking and Optimization Studies

Human milk fat (HMF) triacylglycerols (TAGs) mainly contain palmitic acid esterified at the sn-2 position while oleic and other unsaturated fatty acids are located at positions sn-1,3. This study aimed at the production of HMF substitutes (HMFS) by lipase-catalyzed acidolysis of tripalmitin with oleic acid, in a solvent-free medium. Burkholderia cepacia lipase (BCL) was immobilized in silica (prepared with protic or aprotic ionic liquids) by covalent binding or encapsulation and used as biocatalyst. The supports and immobilized biocatalysts were characterized by FTIR, TGA, and SEM. Molecular docking analysis showed that BCL preferentially attacks oleic acid rather than tripalmitin, due to the lower free energy of hydrophobic binding with this acid (−6.5 kcal·mol−1) than with tripalmitin (5.4 kcal·mol−1). Therefore, the tripalmitin attack by BCL and subsequent HMFS production only occurs after the binding to most of the oleic acid molecules. The highest acidolysis activity was obtained with BCL immobilized by covalent binding in prepared silica with aprotic ionic liquid. A central composite rotatable design, as a function of temperature (58–72 °C) and oleic acid/tripalmitin molar ratio (MR = 2:1–6.8:1), was performed for acidolysis optimization. Under optimized conditions (58 °C and MR = 4:1 or 60 °C and MR = 2:1), the oleic acid incorporation of 28 mol.% was achieved after 48 h.

[1]  De-hua Liu,et al.  Progress and perspectives of enzymatic preparation of human milk fat substitutes , 2022, Biotechnology for Biofuels and Bioproducts.

[2]  X. Zou,et al.  Preparation of Human Milk Fat Substitutes: A Review , 2022, Life.

[3]  Yunjun Yan,et al.  1,3-Dioleoyl-2-palmitoyl glycerol (OPO)-Enzymatic synthesis and use as an important supplement in infant formulas. , 2021, Journal of food biochemistry.

[4]  Á. Lima,et al.  Computational and experimental analysis on the preferential selectivity of lipases for triglycerides in Licuri oil , 2021, Bioprocess and Biosystems Engineering.

[5]  Á. Lima,et al.  Lipase activation by molecular bioimprinting: The role of interactions between fatty acids and enzyme active site , 2020, Biotechnology progress.

[6]  L. Krause,et al.  Continuous flow reactor based with an immobilised biocatalyst for the continuous enzymatic transesterification of crude coconut oil. , 2020, Biotechnology and applied biochemistry.

[7]  Á. Lima,et al.  Optimization of the enzymatic hydrolysis of Moringa oleifera Lam oil using molecular docking analysis for fatty acid specificity , 2019, Biotechnology and applied biochemistry.

[8]  M. Freire,et al.  Enhanced Activity of Immobilized Lipase by Phosphonium-Based Ionic Liquids Used in the Support Preparation and Immobilization Process , 2019, ACS Sustainable Chemistry & Engineering.

[9]  S. Ferreira-Dias,et al.  Production of Human Milk Fat Substitutes by Interesterification of Tripalmitin with Ethyl Oleate Catalyzed by Candida parapsilosis Lipase/Acyltransferase , 2019, Journal of the American Oil Chemists' Society.

[10]  M. Freire,et al.  Effects of phosphonium‐based ionic liquids on the lipase activity evaluated by experimental results and molecular docking , 2019, Biotechnology progress.

[11]  Qingzhe Jin,et al.  Human milk fat substitutes: Past achievements and current trends. , 2019, Progress in lipid research.

[12]  N. Osório,et al.  Structured Lipids for Foods , 2019, Encyclopedia of Food Chemistry.

[13]  Ruann Janser Soares de Castro,et al.  Biocatalytic action of proteases in ionic liquids: Improvements on their enzymatic activity, thermal stability and kinetic parameters. , 2018, International journal of biological macromolecules.

[14]  S. Mattedi,et al.  Lipase Immobilization on Silica Xerogel Treated with Protic Ionic Liquid and its Application in Biodiesel Production from Different Oils , 2018, International journal of molecular sciences.

[15]  M. L. Ferreira,et al.  Burkholderia cepacia lipase: A versatile catalyst in synthesis reactions , 2018, Biotechnology and bioengineering.

[16]  C. Akoh,et al.  Biotechnological and Novel Approaches for Designing Structured Lipids Intended for Infant Nutrition , 2017 .

[17]  Faez Iqbal Khan,et al.  The Lid Domain in Lipases: Structural and Functional Determinant of Enzymatic Properties , 2017, Front. Bioeng. Biotechnol..

[18]  Qianchun Deng,et al.  Lipase immobilized in ordered mesoporous silica: A powerful biocatalyst for ultrafast kinetic resolution of racemic secondary alcohols , 2017 .

[19]  E. Batista,et al.  Applications of Ionic Liquids in the Food and Bioproducts Industries , 2016 .

[20]  N. Osório,et al.  Camelina oil as a source of polyunsaturated fatty acids for the production of human milk fat substitutes catalyzed by a heterologous Rhizopus oryzae lipase , 2016 .

[21]  Fengxiu Zhang,et al.  Key factors affecting the activity and stability of enzymes in ionic liquids and novel applications in biocatalysis , 2015 .

[22]  Wei Wei,et al.  Synthesis of structured lipid 1,3-dioleoyl-2-palmitoylglycerol in both solvent and solvent-free system , 2015 .

[23]  Maria Isabel Rodrigues,et al.  Experimental Design and Process Optimization , 2014 .

[24]  S. Huo,et al.  Enhancing stabilities of lipase by enzyme aggregate coating immobilized onto ionic liquid modified mesoporous materials , 2014 .

[25]  S. Ferreira-Dias,et al.  Human milk fat substitutes: Advances and constraints of enzyme-catalyzed production , 2014 .

[26]  F. Valero,et al.  Production of Human Milk Fat Substitutes Catalyzed by a Heterologous Rhizopus oryzae Lipase and Commercial Lipases , 2014 .

[27]  Roger A Sheldon,et al.  Enzyme immobilisation in biocatalysis: why, what and how. , 2013, Chemical Society reviews.

[28]  G. Zanin,et al.  Protic ionic liquid as additive on lipase immobilization using silica sol-gel. , 2013, Enzyme and microbial technology.

[29]  Bin Zou,et al.  Immobilization of Burkholderia cepacia lipase on functionalized ionic liquids modified mesoporous silica SBA-15 , 2012 .

[30]  G. Sandoval,et al.  Carica papaya latex: A low‐cost biocatalyst for human milk fat substitutes production , 2012 .

[31]  E. Molina,et al.  Enzymatic production of human milk fat substitutes containing palmitic and docosahexaenoic acids at sn-2 position and oleic acid at sn-1,3 positions , 2011 .

[32]  D. Wei,et al.  Carica papaya lipase-catalyzed synthesis of terpene esters , 2011 .

[33]  M. Lepore,et al.  FT-IR microscopy characterization of sol–gel layers prior and after glucose oxidase immobilization for biosensing applications , 2011 .

[34]  M. Ribeiro,et al.  Production of human milk fat substitutes enriched in omega-3 polyunsaturated fatty acids using immobilized commercial lipases and Candida parapsilosis lipase/acyltransferase , 2010 .

[35]  H. Mu.  Production and nutritional aspects of human milk fat substitutes , 2010 .

[36]  Arthur J. Olson,et al.  AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading , 2009, J. Comput. Chem..

[37]  Maggel Deetlefs,et al.  Assessing the greenness of some typical laboratory ionic liquid preparations , 2010 .

[38]  David S. Goodsell,et al.  AutoDock4 and AutoDockTools4: Automated docking with selective receptor flexibility , 2009, J. Comput. Chem..

[39]  I. André,et al.  Insights into lid movements of Burkholderia cepacia lipase inferred from molecular dynamics simulations , 2009, Proteins.

[40]  R. Y. Wei,et al.  Effect of nonsurfactant template content on the particle size and surface area of monodisperse mesoporous silica nanospheres , 2009 .

[41]  Eduardo Filipe,et al.  On the critical temperature, normal boiling point, and vapor pressure of ionic liquids. , 2005, The journal of physical chemistry. B.

[42]  F. F. Moraes,et al.  Studies on immobilizd lipase in hydrophobic sol-gel , 2004 .

[43]  G J Lye,et al.  Room-temperature ionic liquids as replacements for organic solvents in multiphase bioprocess operations. , 2000, Biotechnology and bioengineering.

[44]  Yen Wei,et al.  Preparation and Physisorption Characterization of d-Glucose-Templated Mesoporous Silica Sol−Gel Materials , 1999 .

[45]  T. Welton.  Room-Temperature Ionic Liquids. Solvents for Synthesis and Catalysis. , 1999, Chemical reviews.