IDM ¦ Ingredients
Figure 3:
Free fatty acids formed in brine cheese with lipase from
P. citrinopileatus (PCI_Lip) after 30 days of ripening compared
to the reference enzyme opti-zym z10uc. The peak
areas (measurements in duplicate) determined by means
of gas chromatography with coupled mass spectrometry
after solid phase microextraction (SPME-GC-MS) and
show the different doses of enzyme (2.3 - 15 U/3 L).
38 · March/April 2022 ¦ international-dairy.com
The comparable fatty acid profiles between reference
and fungal lipase are clearly recognizable, which leads
to the characteristic flavor of the cheese. As the
dosage increases, more free fatty acids are produced.
C2: Acetic acid, C4: Butyric acid, C6: Caproic acid,
C8: Caprylic acid, C10: Capric acid, C12: Lauric acid,
C14: Myristic acid, C16: Palmitic acid.
addition to the Enoki mushroom (alias Velvet Shank; Flammulina
velutipes), the golden oyster mushroom (Pleurotus citrinopileatus)
proved to be a promising candidate for the production of extracellular
lipolytic enzymes. It has been shown that these mushrooms
produce lipases that have a similar substrate specificity regarding
the fatty acid chain length as compared to commercially available
animal enzyme preparations (Fig. 1). For this purpose, the mycelium
of edible mushrooms was cultivated in shaking flasks and bioreactors
under defined conditions (Fig. 2). The fungal mycelium is
subsequently separated and discarded, and the enzymes secreted
from the fungus remain in the nutrient medium, from which they
can be isolated and purified by various separation methods. The
fungal lipases were then biochemically characterized at JLU Giessen.
optiferm GmbH carried out application tests with purified lipases
from the culture supernatant, for the production of brine
feta cheese on the basis of cow's milk, as well as with recombinant
variants of these lipases. The cheese produced was subjected to a
gas-chromatographic analysis in Giessen after appropriate ripening
intervals of several months, in order to gain insight into the
free fatty acid profiles formed. In the chromatogram, the reference
cheese produced from goat lipase opti-zym z10uc has a typical
profile, which is formed by the release of short chain fatty acids
from the milk fat, namely butyric acid, caproic acid, caprylic acid
and capric acid (Fig. 3). In parallel, optiferm GmbH carried out sensory
analyses (Fig. 4). To validate the fatty acid profiles and flavor
profiles, also the cheese produced with the reference goat lipase
has undergone tasting. The cheeses made from fungal lipases
showed no significant differences in consistency, texture, color and
aroma when compared to the cheese control group made from
goat lipase. The use of fungal lipases also resulted in very similar
sensory properties, which are also reflected in the free fatty acid
analysis of the cheese. In particular, the use of very low dosages
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