The strophariaceous basidiomycete Cyclocybe aegerita (synonyms Agrocybe aegerita and A. cylindracea) is one of the most praised cultivated edible mushrooms and is being cultivated at large scale for food production. Furthermore, the fungus serves as a model organism to study fruiting body formation and the production of secondary metabolites during the life cycle of Basidiomycota. By studying the secondary metabolite profiles of C. aegerita, we found several terpenoids in submerged cultures. Aside from the main metabolite, bovistol (1), two new bovistol derivatives B and C (2, 3) and pasteurestin C as a new protoilludane (4) were isolated by preparative HPLC. Their structures were elucidated by mass spectrometry and NMR spectroscopy. The relative configurations of 2–4 were assigned by ROESY correlations, and 3
J
H,H coupling constants in the case of 4. Applying quantitative PCR for gene expression validation, we linked the production of bovistol and its derivatives to the respective biosynthesis gene clusters.
Traditionally produced piquant cheeses
such as Feta or Provolone
rely on pregastric lipolytic enzymes of animal origin to intensify
flavor formation during ripening. Herein, we report a novel fungal
lipase, derived from the phylum Basidiomycota to replace animal-derived
products. A screening of 31 strains for the desired hydrolytic activities
was performed, which revealed a promising fungal species. The secretome
of an edible golden oyster mushroom, Pleurotus citrinopileatus, provided suitable enzymatic activity, and the coding sequence of
the corresponding enzyme was identified by combining transcriptome
and liquid chromatography high-resolution electrospray tandem mass
spectroscopy (LC-HR-ESI-MS/MS) data. Recombinant expression in Escherichia coli BL21 (DE3) using chaperones GroES-GroEL
and DnaK-DnaJ-GrpE was established. The recombinant lipolytic enzyme
was purified and biochemically characterized in terms of thermal and
pH stability, optimal reaction conditions, and kinetic data toward p-nitrophenyl esters. An application in the microscale production
of Feta-type brine cheese revealed promising sensory properties, which
were confirmed by headspace solid-phase microextraction gas chromatography
mass spectrometry (HS-SPME-GC–MS) analyses in comparison with
the reference enzyme opti-zym z10uc from goat origin. Supplementation
with 2.3 U of the heterologously expressed fungal lipase produced
the most comparable free fatty acid profile after 30 days of ripening.
The flavor and texture formed during the application of the new lipase
from P. citrinopileatus proved to be
competitive to the use of pregastric lipases and could therefore replace
the products of animal origin.
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