Abstract
Palm stearin fractionate (PSF), obtained from palm stearin by further fractionation with solvents and n-3 polyunsaturated fatty acids (n-3 PUFA) rich fish oil (FO) were subjected to interesterification at 1:1, 1:2, 1:3, 2:1 and 3:1 substrate molar ratio and catalyzed by lipase from Thermomyces lanuginosa for obtaining a product with triacylglycerol (TAG) structure similar to that of human milk fat (HMF). The parameters (molar ratio and time) of the interesterification reaction were standardized. The temperature of 60 °C and enzyme concentration of 10 % (w/w) were kept fixed as these parameters were previously optimized. The reactions were carried out in a stirred tank reactor equipped with a magnetic stirrer for 6, 12, 18 and 24 h. The blends were analyzed for fatty acid (FA) composition of both total FAs and those at the sn-2 position after pancreatic lipase hydrolysis. All the blended products were subjected to melting point determination and free fatty acid content. Finally, blend of PSF and FO at 2:1 molar ratio with 69.70 % palmitic acid (PA) content and 12 h of reaction produced the desired product with 75.98 % of PA at sn-2 position, 0.27 % arachidonic acid (AA), 3.43 % eicosapentaenoic acid (EPA) and 4.25 % docosahexaenoic acid (DHA) and with melting point of 42 °C. This study portrayed a successful preparation of TAG containing unique FA composition i.e. ≥ 70 % of the PA, by weight, were esterified at the sn-2 position which could be used in infant formulation with health benefits of n-3 PUFAs.
Similar content being viewed by others
References
Akoh CC (2006) Handbook of functional lipids. CRC press, Boca Raton
AOCS (1989a) Slip melting point method, Cc 1 - 25. Official methods and recommended practices of the America Oil Chemists Society, 4th edn. AOCS Press, Champaign
AOCS (1989b) Free fatty acids method, Te 1a - 64. Official methods and recommended practices of the America Oil Chemists Society, 4th edn. AOCS Press, Champaign
Aoe S, Yamamura JI, Matsuyama H, Hase M, Shiota M, Miura S (1997) The positional distribution of dioleoyl-palmitoyl glycerol influences lymph chylomicron transport, composition and size in rats. J Nutr 127:1269–1273
Birch EE, Castaneda YS, Wheaton DH, Birch DG, Uauy RD, Hoffman DR (2005) Visual maturation of term infants fed long-chain polyunsaturated fatty acid-supplemented or control formula for 12 mo. Am. J. Clin. Nutr. 81:871–879
Breckenridge WC, Marai L, Kuksis A (1969) Triglyceride structure of human milk fat. Can J Biochem 47(8):761–769
Carnielli VP, Luijendijk IH, Beek RHV, Boerma GJ, Degenhart HJ, Sauer PJ (1995) Effect of dietary triacylglycerol fatty acid positional distribution on plasma lipid classes and their fatty acid composition in preterm infants. Am J Clin Nutr 62(4):776–781
El-Loly MM (2011) Composition, properties and nutritional aspects of milk fat globule membrane – a review. Pol J Food Nutr Sci 61(1):7–32
Fleith M, Clandinin MT (2005) Dietary PUFA for preterm and term infants: review of clinical studies. Crit. Rev. Food Sci 45:205–229
Guesnet P, Alessandri JM (2011) Docosahexaenoic acid (DHA) and the developing central nervous system (CNS) - implications for dietary recommendations. Biochimie 93:7–12
Gunstone FD (2001) Why are structured lipids and new lipids sources required? In: Gunstone FD (ed) Structured and modified lipids. Marcel Dekker, New York, NY, pp. 1–35
Innis SM (2011) Dietary triacylglycerol structure and its role in infant nutrition. Adv Nutr 2(3):275–283
Innis SM, Dyer R, Quinlan P, Diersen-Schade D (1995) Palmitic acid is absorbed as sn-2 monopalmitin from milk and formula with rearranged triacylglycerols and results in increased plasma triglyceride sn-2 and cholesteryl ester palmitatein piglets. J Nutr 125:73–81
Innis SM, Gilley J, Werker J (2001) Are human milk long-chain polyunsaturated fatty acids related to visual and neural development in breast-fed term infants? J Pediatr 139:532–538
Jensen RG (1995) Comments on the extraction of fat from human milk for analysis of contaminants. Chemosphere 31(9):4197–4200(author reply 200-5)
Karabulut I, Turan S, Vural H, Kayahan M (2007) Human milk fat substitute produced by enzymatic interesterification of vegetable oil blend. Food Technol Biotech. 45(4):434–438
Kennedy K, Fewtrell MS, Morley R (1999) Double-blind, randomized trial of a synthetic triacylglycerol in formula-fed term infants: effects on stool biochemistry, stool characteristics, and bone mineralization. Am. J. Clin. Nutr. 70(5):920–927
Koletzko B, Tangermann R, von Kries R, Stannigel H, Willberg B, Radde I, Schmidt E (1998) Intestinal milk-bolus obstruction in formula fed premature infants given high doses of calcium. J Pediatr Gastr Nutr. 7:548–553
Li R, Pande G, Sabir JSM, Baeshen NA, Akoh CC (2014) Enrichment of refined olive oil with palmitic and docosahexaenoic acids to produce a human milk fat analogue. J Am Oil Chem Soc 91(8):1377–1385
Lien EL (1994) The role of fatty acid composition and positional distribution in fat absorption in infants. J. Pediatr 125(5 Pt. 2):S62–S68.
Lien EL, Yuhas RJ, Boyle FG, Tomarelli RM (1993) Corandomization of fats improves absorption in rats. J Nutr 123(11):1859–1867
Lopez A, Bargallo AIC, Folgoso CC (2001) The influence of dietary palmitic acid triacylglyceride position on the fatty acid, calcium and magnesium contents of at term newborn faeces. Early Hum Dev Suppl 65:S83–S94
Luddy FE, Barford RA, Herb SF, Magidman P, Riemenschneider RW (1963) Pancreatic lipase hydrolysis of triglycerides by a semi micro technique. Presented in part at the AOCS Meeting, Minneapolis
Metcalfe LD, Schmitz AA (1961) The rapid preparation of fatty acid esters for gas chromatographic analysis. J Anal Chem 33(3):363–364
Mu H, Hoy HE (2004) The digestion of dietary triacylglycerols. Prog Lipid Res 43(2):105–133
Reyes HR, Hill CG (1994) Kinetic modeling of interesterification reactions catalyzed by immobilized lipase. Biotechnol Bioeng 43(2):171–182
Sellappan A, Akoh CC (2001) Synthesis of structured lipids by transesterification of trilinolein catalyzed by lipozyme IM60. J Agric Food Chem 49:2071–2076
Shimada Y, Nagao T, Hamasaki Y, Akimoto K, Sugihara A, Fujikawa S, Komemushi S, Tominaga Y (2000) Enzymatic synthesis of structured lipid containing arachidonic and palmitic acids. J Am Oil Chem Soc 77:89–93
Squire TL, Lowe ME, Bauer VW, Andrews MT (2003) Pancreatic triacylglycerol lipase in a hibernating mammal. II. Cold-adapted function and differential expression. Physiol Genomics Dec 16; 16 (1):131–140.
Teichert SA, Akoh CC (2011) Stearidonic acid soybean oil enriched with palmitic acid at the sn-2 position by enzymatic interesterification for use as human milk fat analogues. J Agric Food Chem 59:5692–5701
Turan D, Yesilcubuk NS, Akoh CC (2012) Production of human milk fat analogue containing docosahexaenoic and arachidonic acids. J Agric Food Chem 60:4402–4407
Wang ZJ, Liang CL, Li GM, Yu CY, Yin M (2006) Neuroprotective effects of arachidonic acid against oxidative stress on rat hippocampal slices. Chem Biol Interact 163(3):207–217
Xu X (2000) Production of specific-structured triacylglycerols by lipase-catalyzed reactions: a review. Eur J Lipid Sci Technol 102:287–303
Zou XQ, Huang JH, Jin QZ, Liu YF, Song ZH, Wang XG (2011) Lipase-catalyzed preparation of human milk fat substitutes from palm stearin in a solvent-free system. J Agric Food Chem 59:6055–6063
Zou XQ, Huang JH, Jin QZ, Liu YF, Tao GJ, Cheong LZ, Wang XG (2012a) Preparation of human milk fat substitutes from palm stearin with arachidonic and docosahexaenoic acid: combination of enzymatic and physical methods. J Agric Food Chem 60:9415–9423
Zou XQ, Guo Z, Huang JH, Jin QZ, Cheong LZ, Wang XG, Xu XB (2012b) Human milk fat globules from different stages of lactation: a lipid composition analysis and microstructure characterization. J Agric Food Chem 60(29):7158–7167
Zou X, Huang J, Jin Q, Liu Y, Song Z, Wang X (2012c) Lipase-catalyzed synthesis of human milk fat substitutes from palm stearin in a continuous packed bed reactor. J Am Oil Chem Soc 89(8):1463–1472
Acknowledgment
The financial assistance received from Department of Science and Technology (DST), Govt. of India by providing INSPIRE fellowship to Ms. Moumita Ghosh and UGC, Govt. of India by providing DSK Kothari Post Doctoral Fellowship to Dr. Avery Sengupta, are gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Additional information
Research Highlights:
1. Human milk fat has unique composition of sn-2 position which is enriched with palmitic acid. Various attempts to produce human milk fat analogue are taking place world wide.
2. Enzymatic reactions are essentially suitable for this reactions which deals with heat sensitive starting materials.
3. Palm stearin, a by product of palm oil industry, was utilised in this study to prepare human milk fat analogue. To provide the essential fatty acids, especially DHA, fish oil was also used.
4. On enzymatic interesterification, specific blends of palm stearin fractionate and fish oil produced human milk fat analogue with desired fatty acid composition and melting point.
5. The study highlighted the utilisation of one by product to produce value added product.
Rights and permissions
About this article
Cite this article
Ghosh, M., Sengupta, A., Bhattacharyya, D.K. et al. Preparation of human milk fat analogue by enzymatic interesterification reaction using palm stearin and fish oil. J Food Sci Technol 53, 2017–2024 (2016). https://doi.org/10.1007/s13197-016-2180-5
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13197-016-2180-5