Oleaginous fungi are efficient tools to convert agricultural waste streams into valuable components. The filamentous fungus Mucor circinelloides was cultivated in whey permeate, a byproduct from cheese production, to produce an oil-rich fungal biomass. Response surface methodology was used to optimize the fermentation conditions such as pH and temperature for increased biomass yield and lipid accumulation. Quantification and characterization of the fungal biomass oil was conducted. Upstream lactose hydrolysis of the whey permeate increased the biomass yield from 2.4 to 7.8 (g dry biomass/L) compared to that of non-hydrolyzed whey permeate. The combination of low pH (4.5) and pasteurization minimized microbial competition, thus favoring fungal growth. A central composite rotatable design was used to evaluate the effects of temperature (22.4–33.6 °C) and a lower pH range (3.6–4.7) on biomass yield and composition. The highest biomass yield and oil content was observed at high temperature (33.6 °C), while the pH range evaluated had a less pronounced effect. The predictive model was validated at the optimal conditions of 33.6 °C and pH 4.5. The fungal biomass yield plateaued at 9 g dry cell weight per liter, while the oil content and lipid yield reached a maximum of 24% dry biomass and 2.20 g/L, respectively, at 168 h. Triacylglycerides were the major lipid class (92%), which contained predominantly oleic (41%), palmitic (23%), linoleic (11%), and γ-linolenic acid (9%). This study provided an alternative way of valorization of cheese whey permeate by using it as a substrate for the production of value-added compounds by fungal fermentation. The fatty acid profile indicates the suitability of M. circinelloides oil as a potential feedstock for biofuel production and nutraceutical applications.

This work investigates the composition of whey protein phospholipid concentrate (WPPC), an underutilized dairy stream, and reveals that it is a source of many bioactive compounds that can benefit the immune system and gut health. Our glycoproteomics approach uncovered that proteins derived from the milk fat globule membrane (MFGM) represent 23% of the total protein relative abundance and identified 85 N-glycans. Released sialic acid, an additional marker of glycosylation, ranged from 1.2 to 2% of the total weight. Xanthine oxidase, a glycosylated marker of MFG bioactivity, was found in high abundance and displayed higher antimicrobial activity than bovine milk, despite its similar fat and solids content. An average MFG diameter of 2.64 ± 0.01 µm was found in liquid WPPC, compared to 4.78 ± 0.13 µm in bovine milk, which likely explains the unusually high presence of glycosylated membrane-bound proteins and phospholipids, whose total fatty acids accounted for 20% of the WPPC total fatty acid pool. Free and bound oxylipins (mainly derived from linoleic acid) were also identified, together with other less abundant anti-inflammatory lipid mediators derived from eicosapentaenoic acid and docosahexaenoic acid. Our study demonstrates that WPPC represents a promising starting material for bioactive compound extraction and a functional vehicle for the delivery of small MFGs.

Human milk provides bioactive compounds such as milk fat globules (MFGs), which promote brain development, modulate the immune system, and hold antimicrobial properties. To ensure microbiological safety, donor milk banks apply heat treatments. This study compares the effects of heat treatments and homogenization on MFG’s physicochemical properties, bioactivity, and bioavailability. Vat pasteurization (Vat-PT), retort (RTR), and ultra-high temperature (UHT) were performed with or without homogenization. UHT, RTR, and homogenization increased the colloidal dispersion of globules, as indicated by increased zeta potential. The RTR treatment completely inactivated xanthine oxidase activity (a marker of MFG bioactivity), whereas UHT reduced its activity by 93%. Interestingly, Vat-PT resulted in less damage, with 28% activity retention. Sialic acid, an important compound for brain health, was unaffected by processing. Importantly, homogenization increased the in vitro lipolysis of MFG, suggesting that this treatment could increase the digestibility of MFG. In terms of color, homogenization led to higher L* values, indicating increased whiteness due to finer dispersion of the fat and casein micelles (and thus greater light scattering), whereas UHT and RTR increased b* values associated with Maillard reactions. This study highlights the nuanced effects of processing conditions on MFG properties, emphasizing the retention of native characteristics in Vat-PT-treated human milk.