Abstract:Yarrowia lipolytica YlTun15 (GeneBank Acc. N° MF327143), isolated from farmed Dicentrarchus labrax's gills, secretes an alkaline extracellular protease, which exhibited the highest activity at pH 9 and temperature 45°C. The enzyme activity extracted was tested in the presence of different ion metal and protein inhibitors. .48% respectively at the 5 mM level. The enzyme was almost (activity=1.47%) inhibited by phenylmethylsulfonyl fluoride at the concentration of 5 mM. However, the protease activity was relatively constant in the presence of EDTA and SDS that may conclude that this enzyme was not a metalloprotease and belong to the serine protease category. After 18 months-storage at -20°C, the enzyme activity has decreased to 23.17%. This protease may have a potential application in food and detergent activity.

Background: Among the third-generation biodiesel feed stock, oleaginous marine yeasts are the least studied microorganisms for such purpose.Results: Wild strains yeasts were isolated from various Tunisian marine sources including fish waste (Candida tenuis CtTun15, Debaryomyces hansenii DhTun2015, Trichosporon asahii TaTun15 and Yarrowia lipolytica YlTun15) and seawater (Rhodotorula mucilaginosa RmTun15). Following incubation with ethyl methanesulfonate (EMS: 75 mM) for various periods of time (T15, T30, T45, T60 min), the cell viability of these strains responded differentially according to yeast species. For instance, mutated CtTun15 did not survive after 30 min of EMS treatment; higher resistances were observed in DhTun2015 (45 min), in YlTun15, RmTun15 and in TaTun15 (60 min) but with significant decreased cell viabilities (survival rate: 6.02, 3. 16, 11.22, 11.58, 7.70%, respectively). For all surviving mutated strains, the optima of biomass and lipid yields were detected after 96 h in YPD culture; but derived from strains submitted to different period of EMS incubation. In most mutated strains, the maximum biomass (BP) and lipid (LP) productivities coincided and were observed after 30 min of EMS incubation. Only CtTun15 showed different optima of BP and LP (after 30 min and 15 min, respectively). The fatty acids (FA) compositions considered essential in the prediction of biodiesel criteria; were highly affected by EMS mutagenesis. Essentially, 30-and 45-min EMS incubation induced the highest levels of PUFA and MUFA in YlTun15, RmTun15 and TaTun15 with non-significant differences in the different times. However, CtTun15 and DhTun2015 mutant strains responded differently, with the highest levels of MUFA observed following 15 and 45 min; and that of PUFA after 30 and 45 min, respectively. Conclusion:The methyl-esterification of FA from the three mutated yeast strains (30 min-YlTun15, RmTun15 and TaTun15) yielded biodiesel with physical proprieties consistent with the International Standard System. However, investigations are needed for up-scaling biodiesel production.

Cell-based agriculture is an emerging and attractive alternative to produce various food ingredients. In this study, five strains of marine yeast were isolated, molecularly identified and biochemically characterized. Molecular identification was realized by sequencing the DNA ITS1 and D1/D2 region, and sequences were registered in GenBank as Yarrowia lipolytica YlTun15, Rhodotorula mucilaginosa RmTun15, Candida tenuis CtTun15, Debaryomyces hansenii DhTun2015 and Trichosporon asahii TaTun15. Yeasts showed protein content varying from 26% (YlTun15) to 40% (CtTun15 and DhTun2015), and essential amino acids ranging from 38.1 to 64.4% of the total AAs (CtTun15-YlTun15, respectively). Lipid content varied from 11.15 to 37.57% with substantial amount of PUFA (>12% in RmTun15). All species had low levels of Na (<0.15 mg/100 g) but are a good source of Ca and K. Yeast cytotoxic effect was investigated against human embryonic kidney cells (HEK 293); results showed improved cell viability with all added strains, indicating safety of the strains used. Based on thorough literature investigation and yeast composition, the five identified strains could be classified not only as oleaginous yeasts but also as single cell protein (SCP) (DhTun2015 and CtTun15) and single cell oil (SCO) (RmTun15, YlTun15 and TaTun15) producers; and therefore, they represent a source of alternative ingredients for food, feed and other sectors.

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