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Proceeding of the 4 rd International Symposium“NEW RESEARCH IN BIOTECHNOLOGY” USAMV Bucharest, Romania, 20113. Study of the nitrogen source influenceIn a first step, growth and carotenoids synthesis, including the torularhodin formation withthe Rhodotorula rubra yeast were investigated based on the use of different salts asmineral nitrogen sources such as NH 4 NO 3 , NH 4 H 2 PO 4 , (NH 4 ) 2 SO 4 , NH 4 Cl and NH 4 NO 3 (asthe control salt initially present in MS3 medium) with the concentration of 5 g/L for all thevariants. At the same time for the control medium composition variant the organic nitrogensource, the yeast extract, was supplemented with a concentration of 0.1% from thefollowing amino acids: tryptophan, threonine, glutamic acid, cysteine, alanine, tyrosine,proline, leucine and valine. The experiment lasted for 144 h.The pigments formation results are presented in the fig. 4.Table 1. Experimental variants notation function of the use of different nitrogen sourcesExperimentalNitrogen source,variants5 g/L1 NH 4 NO 3 - control2 NH 4 H 2 PO 43 (NH 4 ) 2 SO 44 NH 4 Cl5 NH4NO3 + 0.1% tryptophan6 NH4NO3 + 0.1% threonine7 NH4NO3 + 0.1% glutamic acid8 NH4NO3 + 0.1% cysteine9 NH4NO3 + 0.1% alanine10 NH4NO3 + 0.1% tyrosine11 NH4NO3 + 0.1% proline12 NH4NO3 + 0.1% leucine13 NH4NO3 + 0.1% valine900m800iued 700m600/L500gµ 400,n 300tio200atr 100cen0noC1 2 3 4 5 6 7 8 9 10 11 12 13Experimental variantsTotal carotenoids concentration as ß-carotene Torularhodin concentrationFig. 4 Variation of the torularhodin concentration, and the total carotenoids concentration (as β-carotene) depending of the nitrogen source from the brothCompared to control medium (MS3) all the variants characterized by small additions ofamino acids have better growths. The introduction of alanine and threonine seems to100
Proceeding of the 4 rd International Symposium“NEW RESEARCH IN BIOTECHNOLOGY” USAMV Bucharest, Romania, 2011influence more the pigments formation than the use of NH 4 H 2 PO 4 instead of the controlmineral nitrogen source.4. Influence of fatty acids additionsIn order to study the possible influence of the introduction of small concentrations of fattyacids on growth and pigments formation considering the Rhodotorula yeasts capacity toassimilate these acids as valuable carbon sources [11] the control medium formula wassupplemented with 0.1% of fatty acids: linoleic/oleic/stearic/palmitic. Again the growth andthe total carotenoids concentration compared with the torularhodin concentration weredetermined. The carotenoids formation, including torularhodin is presented in the Table 2,and the growth curves for the same experimental variants are represented in the fig. 5.Table 2. Experimental variants notation function of the use of fatty acidsCLOSPMediumcompositionvariantsFatty acids additionControl (MS3)Glucose + 0.1% linoleic acidGlucose + 0.1% oleic acidGlucose + 0.1% stearic acidGlucose + 0.1% palmitic acidThe growth was better when oleic acid was used and the carotenoids and torularhodinconcentrations are the highest for this variant.It is to mention that the influence of the addition of small concentrations of amino acidsalanine or threonine was more important.2018mn0y,6sitenaldticpO16141210864200 20 40 60 80 100 120 140 160Time, hControl L O S PFig. 5 Influence of fatty acids addition on the growth of Rhodotorula rubra ICCF 209 yeast101
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