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Importantly, the intracellular PHB concentration (%PHB/CDW) was 2.7-fold higher in the medium containing nitrogen than in the nitrogen deficient medium (Fig. 3; Table 1).
Similar(59)
The medium used in this experiment and routinely for C. thermocellum growth contains a large excess of nitrogen source and it is possible to culture C. thermocellum in medium containing less nitrogen [ 41].
We confirmed this finding for cells grown in minimal media with defined nitrogen source; however, when cells were grown in rich yeast extract medium containing complex nitrogen sources, Sck2p became important for normal levels of both polysomes and S6 phosphorylation (Fig. 5D,E).
Therefore, Asther medium containing all nitrogen sources was used in our further studies.
When pdh1-1, pdh1-4 and p5cdh-2 mutants were grown on medium containing mineral nitrogen, only p5cdh-2 was more sensitive to growth-inhibition by external Pro than the wildtype.
Human CBS function was assayed based upon the growth of cys4Δ yeast strains in liquid and on solid minimal medium containing Yeast Nitrogen Base without vitamins (Qbiogene, Calsbad, CA) or GSH.
While performing phenotypic analysis of the phosphatase mutants, we observed that a PP2C class phosphatase knockout mutant (Δ pph-8) displayed inappropriate protoperithecial formation on VM medium containing high nitrogen.
For the growth analysis, recombinant yeast cells were grown – at 28°C in a synthetic Trp-free liquid medium containing yeast nitrogen base and required amino acids – up to reach a mid-log phase.
More than half the acylglycerols were triacylglycerols.> -wrap-foot> aBorra = spent coffee grounds bYNBO = minimal oleic acid medium containing yeast nitrogen base cOn various glucose, glycerol, and volatile fatty acid combination substrates In plate assays, the mutant strains grew on a variety of substrates similar to the wild-type strain as shown in Fig. 7.
The surfaces of all cells in strain F were very bumpy, possibly caused by numerous oil droplets adhering to the surface as suggested by its high oil content, similar to the scanning electron micrographs of Y. lipolytica cells grown in minimal oleic acid medium containing yeast nitrogen base (without amino acids and ammonium sulfate -NH4Cl-yeast extract-oleic acid-50 mM phosulfate -NH4Cl-yeast.
In this scenario, the use of mixture design to develop a culture medium containing a cheaper nitrogen source seems to be more appropriate and simple.
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