Milk Fat Composition

Milk fat is composed primarily of triglycerides (or triacylglycerides) are 98% of the total milk fat
(by weight). Other milk lipids (as a percentage of the total milk fat) include: diacylglycerides
(0.25-0.48%); monoacylglycerides (0.02-0.04%); phospholipids (0.6-1.0%); cholesterol (0.2-
0.4%); glycolipids (0.006%); and free fatty acids in milk (0.1-0.4%).

The fatty acid composition of milk varies substantially among mammalian species.

Sources of Fatty Acids

Milk fat triglycerides are synthesized in the mammary epithelial cells. However, the fatty acids
used to synthesize the milk triglycerides may arise from two sources:

1. from breakdown of blood lipids

2. from de novo synthesis within the mammary epithelial cells.

1. Blood Lipids -

From 40 to 60% of the fatty acids come from the blood. These are primarily derived from very
low density lipoproteins (VLDL), which are synthesized in the intestine or liver. VLDL are
composed of 90 to 95% lipid (55-60% triglyceride) on the inner core and 5 to 10% protein at the
outer surface. Chylomicrons, containing ingested fatty acids from the intestine, also can act as a
source of blood-derived fatty acids for the mammary gland.

Triglycerides in the VLDL are hydrolyzed in the mammary capillaries by an enzyme

called lipoprotein lipase (LPL). The LPL can hydrolyze off one, two or all three of the fatty acids
from the glycerol backbone, resulting in free fatty acids plus diacylglycerides,
monoacylglycerides, or glycerol, respectively. The free fatty acids, monacylglycerides,
diacylgycerides and glycerol can all be taken up by the mammary epithelial cell and reused for
triglyceride synthesis.

The FAs contained in VLDL and cylomicrons are dependent upon dietary lipids and on
mobilized fat from body adipose. In the nonruminant, the FA composition of the diet can directly
affect the FA composition of the milk. Feeding supplemental dietary fat can increase milk fat
yield and FA composition of the milk fat. However, in ruminants, diets are typically low in
dietary lipid and that lipid is metabolized by the rumen. The result is that cow milk FA
composition is not normally regulated much by diet. However, in cases of bypass fat in diets of
ruminants the lipids pass directly to the intestine and become part of the FA profile of the VLDL
and chlyomicrons. Cow milk FA composition can be altered by bypass dietary lipid.

2. De novo Fatty Acid Synthesis -

Synthesis of short and medium chain fatty acids in the mammary gland occurs by de novo
synthesis (synthesis from the start, or synthesis of new molecules of fatty acids from precursors
absorbed from the blood). De novo synthesis of fatty acids occurs in the cytoplasm of the
mammary epithelial cell. In the ruminant, the carbon sources used for FA synthesis are acetate
(the most important one) and b-hydroxybutyrate (BHBA). Glucose is a carbon source for FA
synthesis in nonruminants. The reducing equivalents needed for FA synthesis come from
NADPH2 (nicotinamide adenine dinucleotide phosphate, reduced form). The two key enzymes
involved in fatty acid synthesis in the mammary gland are:

Acetyl-CoA Carboxylase, which is the rate limiting enzyme for the fatty
acid synthesis pathway.
Fatty Acid Synthetase, which is a large complex of enzyme activities
responsible for the chain elongation of the fatty acid chains.

The relative proportions of milk fatty acids derived from de novo synthesis or blood lipids is
dependent on carbon chain length of the fatty acid. Typically, shorter chain fatty acids arise
predominantly from de novo synthesis in the epithelial cell, while longer chain fatty acids arise
directly from blood lipids.

Triglyceride Synthesis

Synthesis if triglycerides from FAs (preformed from the blood or synthesized in the cell by de
novo synthesis) occurs at the cytoplasmic surface of the smooth endoplasmic reticulum (SER).
The FAs are esterified to the hydroxyl groups of the glycerol molecules. This occurs by a series
of esterases. There is some specificity of which carbon on the glycerol is used to esterify
particular chain length FAs. In the cow, C16 is found predominantly on the #2 carbon of
glycerol, C18 FAs mostly on the #3 carbon, C4-C8 mostly on the #3 carbon, and C10-C14
distributed among each of the glycerol carbons.

Lipid Droplet Formation

Inside the mammary epithelial cell, as the triglycerides are synthesized at the outer surface of the
SER (smooth endoplasmic reticulum), they start coalescing and form micro lipid droplets. These
micro lipid droplets bud or bleb off from the SER surface into the cytoplasm. The micro lipid
droplets may be secreted from the cells directly as very small milk fat globules (less than 0.5
mm), they may fuse with each other to form larger droplets (cytoplasmic lipid droplets), and they
may fuse with cytoplasmic droplets , ultimately resulting in formation of larger milk lipid
droplets. Milk fat globules (in milk) range from less than 0.5 to greater than15 mm. The micro
and cytoplasmic lipid droplets are not surrounded by a lipid bilayer membrane but apparently are
surrounded by a nonbilayer coating made of protein and polar lipid (gangliosides). This surface
coating prevents coalescence of the droplets with lipids in the cell, yet permits fusion between
droplets. In fact, the protein and ganglioside of this coating, along with calcium, are involved in
the fusion of the droplets.

Secretion of Milk Fat

As the large milk lipid droplets migrate to the apical surface of the cell they continue to push
outward and are enveloped by the apical membrane of the cell. This membrane eventually fuses
at the base, releasing the membrane-bound milk fat globule, and closing the cell's apical
membrane so ere is no hole in it. So, the milk fat globule is membrane-surrounded and has a
number of membrane-associated proteins. These proteins and others trapped during the process
of separating cream from whole milk (usually by some type of centrifugation) are important for
the whipping properties of cream.