AOAC SPIFAN Stakeholder Panel Meeting Book (September 7, 2019)

HMO and infant formula • Scientific proven HMO health benefits to neonates • Infant formula (IF) based on bovine milk, however bovine MO concentrations are very low • Fortification of infant formula with HMO to make IF closer-like to human milk Challenge • Access to high quantities of HMO needed for IF fortification • Limited number of HMOs available in large scale ¾ 2’FL; 3FL; DFL

Compositionof Bovine milk

Protein(34g/L)

Fat(40g/L)

Lactose (48g/L)

O S (<0.1 g/L)

¾ LNnT; LNT ¾ 3’SL; 6’SL

Methods for HMO synthesis Method Advantages

Challenges

• Contains “authentic” HMO

• Scale-up limited due to limited availability of human donor milk

Isolation from human milk

• Number of MO identical to HMO • Scale-up possible because of available large volumes • BMO as precursors for chemoenzymatic synthesis

• OS very low in bovine milk • Not all HMO present (e.g., only trace amounts of fucosylated OS in bovine milk) • Contain nonhuman OS (safety issues?) • Laborious and time-consuming due to high number of protecting group manipulations • Toxic reagents involved • Not cost-efficient for scale-up

From dairy streams

• Clearly defined structures

Chemical synthesis

• Clearly defined structures and HMO libraries (valuable for structure-function relationships and research) • Scale-up possible and almost unlimited

• Challenging scale-up due to low yields reported so far

Chemoenzymatic synthesis

• Produces a limited set of HMOs at a time • Use of genetically engineered organisms

Microbial engineering

Bode et al. Nutr. Rev. (2016) 74 : 635-644 Zeuner et al. Molecules (2019) 24 : 2033-2054