The Enzymatic Preparation of Human Milk Fat Substitute Intermediate Rich in Palmitic Acid at sn-2 Position and Low-Unsaturated Fatty Acids at sn-1(3) Positions from Palm Oil Substrate

J Oleo Sci. 2021 Feb 1;70(2):165-173. doi: 10.5650/jos.ess20101. Epub 2021 Jan 15.

Abstract

The lipid products that consist of structured lipids rich in palmitic acid (16:0) at the sn-2 position of triacylglycerol (TAG) and rich in low-unsaturated fatty acids (FAs) (LUFAs), such as oleic acid; 18:1 and linoleic acid; 18:2 at the sn-1(3) positions, are useful intermediates for manufacturing human milk fat substitute (HMFS), which contains functional lipid components. In this study, the HMFS intermediate (HMFS-IM) was enzymatically prepared from palm oil without using other oil sources. First, the amount of 16:0 at the sn-2 position of TAG substrate was enhanced from 18.9% to more 34.5% via a random esterification reaction using a non-stereospecific lipase, Novozym® 435, to produce a random-palm substrate. Consequently, 2-monoacylglycerol (2-MAG) rich in 16:0 at the sn-2 position over 88%, together with the FA ethyl ester substrates rich in LUFAs, such as 18:1-Et and 18:2-Et above 93.5% was prepared through ethanolysis reaction using the same lipase from the random-palm substrate and by purification with urea complexation, respectively. As the preferred modified method, a continuous use of the same lipase to these reactions were achieved while reducing the usage of enzyme to half. Finally, an HMFS-IM rich in 16:0 at the sn-2 position more than 60% and LUFA at sn-1(3) positions was prepared using these palm oil-based products, including random-palm, palm-Et, and 2-MAG, via the interesterification reaction using a 1,3-stereospecific lipase, Lipozyme® RM-IM. Thus, HMFS-IM was successfully prepared by palm oil materials with a 65 wt% usage ratio. The concept described in this study will be useful for HMFS manufacturing from a single natural oil substrate, which is not initially rich in 16:0 at the sn-2 position.

Keywords: enzymatic preparation; human milk fat substitute; lipase; palm oil.

MeSH terms

  • Enzymes, Immobilized
  • Esterification
  • Fatty Acids, Unsaturated / chemistry*
  • Fungal Proteins
  • Glycolipids / chemistry*
  • Glycoproteins / chemistry*
  • Linoleic Acid / chemistry
  • Lipase / chemistry
  • Lipid Droplets / chemistry*
  • Milk Substitutes / chemical synthesis*
  • Milk, Human / chemistry*
  • Oleic Acid / chemistry
  • Palm Oil / chemistry*
  • Palmitic Acid / chemistry*
  • Triglycerides / chemistry

Substances

  • Enzymes, Immobilized
  • Fatty Acids, Unsaturated
  • Fungal Proteins
  • Glycolipids
  • Glycoproteins
  • Triglycerides
  • milk fat globule
  • Oleic Acid
  • Palmitic Acid
  • Palm Oil
  • Linoleic Acid
  • Novozyme 435
  • Lipase