Yeast dialysate

Kenny (1973) recommends a Soy peptone-yeast dialysate medium (Appendix 4) for culture of mycoplasma. Inclusion of arginine (16 mM) and 0.4 mg% phenol red indicates the presence of arginine deaminase by formation of alkali (purple coloration). Alternatively, incubation with tritiated thymidine and analysis of the culture medium for tritiated thymine can be used to detect thymidine phosphorylase (House and Waddell, 1967). 

When using purified triolein, most samples are amenable to bioassay after di-alytic enrichment. For example, Microtox bioassay of dialysates of SPMDs shows that the SPMDs made with the purified triolein have lower acute toxicities than dialysates from SPMDs made from unpurified triolein (Johnson, 2001). Finally, examination of the dialysates using the yeast estrogen screen (YES) assay (Routledge and Sumpter, 1996) demonstrated that the purification procedure removes all background estrogenic activity (Lebo et ah, 2004). Use of triolein purified by this process expands the potential applicability of SPMD sample extracts to include numerous bioassay procedures (see Chapter 6) and GC-MS as a standard analysis technique. 

One of the common side effects observed during extractive bioconversion is the accumulation of unwanted by-products in the system which may affect the productivity during continuous operation (14). The build up of glycerol and other non-volatile products was shown to decrease the ethanol yields during repeated fermentations in a two-phase system (12). The problem was, however, solved by dialysing the fermentation broth and also adding more yeast cells. It appears that the combination of ultrafiltration with the phase system may circumvent the problem of by-product inhibition in most of the cases. 

Calcium chromate has been shown to induce cytoplasmic petite mutations in mitochondria of Saccharomyces cerevisiae K Calcium chromate also dramatically depressed the content of the mitochondrial gene products cytochrome aa3 and cytochrome b, in whole yeast cells. Chromate ( 8 nM) was readily taken up by rat thymocytes and after 30 min 9% of the Cr was found in the mitochondria although 62% was found in the nuclei . Isolated rat thymus mitochondria and nuclei readily took up CrOj . After one hour incubation of Erlich ascites tumor cells with CrOj (380 /nuclear fraction and 12% was in the mitochondrial-microsomal fraction. Levels of chromium in rat liver mitochondria reached a plateau six hours after i.v. injection of chromate (0.02 mg/kg) and remained at that level through 5 days. Liver nuclear chromium levels in the same animals, although similar to mitochondrial levels at 6 h, reached a maximum at 12 h and steadily decreased after that time. Therefore the nuclear chromium levels were lower than the mitochondrial chromium levels at later times (24-120 h) after injection. The subcellular distribution of chromium in the liver of rats injected i.v. with chromate (0.56 mg/kg) was also found to be time dependent in another study. The distribution of chromium in rat liver mitochondria increased from 5% at 15 min to 21% at 72 h and also increased in the nuclear fraction from 22% at 15 min to 52% at 72 h. Incubation of isolated rat liver mitochondria with chromate (0.3-16.6 electron transport chain of the mitochondrial iner membrane. 

Heat shock and ethanol treatment. Reactivative action of the dialysate was demonstrated in E. coli, S. cerevisiae and C. guilliermondii subjected to heat shock (Table 2.17). The efficiency of the reactivation was inversely proportional to the viability of the inactivated cells, thus replicating the regularity detected for UV-irradiated bacteria and yeasts. Moreover, the two dialysate fractions that showed reactivative and protective activity in UV-irradiated E. coli also showed reactivative effects in bacterial cells inactivated by heating. 

Dialysed extracts of muscle, liver, brain and yeast contain an enzyme that phosphorylates each glucose unit in glycogen and thus disrupts the polysaccharide into glucose-l-phosphate. 

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