The maltose permease family of Saccharomyces cerevisiae comprises five proteins, three of which are characterized, MAL31, MAL61 and AGT1 and two putative permeases, YDL247w (MPH2 ) and YJR160c (MPH3 ). The two uncharacterized permeases share 100% identity and have 75% identity with MAL31 and MAL61 and 55% identity with AGT1. Characterization of the genes YDL247w and YJR160c confirmed that they encode α-glucoside permeases capable of transporting maltose, maltotriose, α-methylglucoside and turanose. Analysis of the promoter regions identified regulatory elements, binding sites for the transcriptional activator, Malx3p and the inhibitory protein, Mig1p. Further analysis of the flanking sequences located blocks of identity covering five open reading frames, indicating that this region was involved in chromosomal block duplication. The members of the maltose permease family are proteins that have strongly overlapping but nevertheless distinct functions, which is a selective advantage for yeast, as it reflects successful adaptation to the variety of environmental conditions to which the yeast cells are exposed; such adaptability is very important in an industrial context.
Efficient fermentation of maltotriose is a desired property of Saccharomyces cerevisiae for brewing. In a standard wort, maltotriose is the second most abundant sugar, and slower uptake leads to residual maltotriose in the finished product. The limiting factor of sugar metabolism is its transport, and there are conflicting reports on whether a specific maltotriose permease exists or whether the mechanisms responsible for maltose uptake also carry out maltotriose transport. In this study, radiolabeled maltotriose was used to show that overexpression of the maltose permease gene, MAL61, in an industrial yeast strain resulted in an increase in the rate of transport of maltotriose as well as maltose. A strain derived from W303-1A and lacking any maltose or maltotriose transporter but carrying a functional maltose transport activator (MAL63) was developed. By complementing this strain with permeases encoded by MAL31, MAL61, and AGT1, it was possible to measure their specific transport kinetics by using maltotriose and maltose. All three permeases were capable of high-affinity transport of maltotriose and of allowing growth of the strain on the sugar. Maltotriose utilization from the permease encoded by AGT1 was regulated by the same genetic mechanisms as those involving the maltose transcriptional activator. Competition studies carried out with two industrial strains, one not containing any homologue of AGT1, showed that maltose uptake and maltotriose uptake were competitive and that maltose was the preferred substrate. These results indicate that the presence of residual maltotriose in beer is not due to a genetic or physiological inability of yeast cells to utilize the sugar but rather to the lower affinity for maltotriose uptake in conjunction with deteriorating conditions present at the later stages of fermentation. Here we identify molecular mechanisms regulating the uptake of maltotriose and determine the role of each of the transporter genes in the cells.The three principal sugars available to Saccharomyces cerevisiae during the fermentation of alcoholic beverages are glucose, maltose, and maltotriose. Efficient fermentation requires the rapid and complete utilization of all of these sugars (18). Maltotriose, which is the second most abundant sugar in wort, is formed from the breakdown of complex sugars during mashing (19). A common problem in the brewing industry is incomplete utilization, resulting in a beer with residual maltotriose. The incomplete fermentation represents a waste of fermentable extract and a loss in revenue and leads to a product with higher carbohydrate levels and potentially an atypical flavor profile (36). Explanations for poor maltotriose utilization are varied; Findlay (12) believed that maltotriose was not a viable carbon source for brewing strains, whereas Casey et al. (4) suggested that with the advent of high-gravity brewing, increased stress on the yeast cells together with poor nutritional supplementation led to less efficient fermentation. Hornsey (18) proposed that the mutation...
Hypoxanthine transport in the insect trypanosome, Crithidia luciliae, was activated in purine depleted conditions. The existence of 2 hypoxanthine transport mechanisms was established. The first, a non-saturable diffusion system, present in purine replete conditions, exhibited properties that were different from the second transport system which was evident only during purine depleted conditions (purine stress). The rate of transport under purine stress was elevated approximately 8-fold over that in replete conditions. This transporter was saturable with a Km of 3·9 μM for hypoxanthine. The transporter substrate specificity included other purine bases, e.g. adenine and guanine, and the purine nucleoside, adenosine. These inhibited hypoxanthine transport competitively with Ki values of 2 μM, 3 μM and 42 μM respectively. Coincident with the increase of hypoxanthine transport under purine stress, the transport of adenosine increased 4-fold and the activity of the 3′-nucleotidase ectoenzyme also increased significantly. Under purine stress the concurrent increase of hypoxanthine and adenosine transport and the increase in 3′-nucleotidase activity could be repressed with either the supplementation of excess purines or by cycloheximide. This study of purine salvage mechanisms in Crithidia luciliae illustrates the successful adaptation of the parasite to nutritional insufficiency.
View related articles Citing articles: 1 View citing articles MARCH, 19n TEACHING OF PLACE CEOCRAPHY CONCLUSION And thus was our atlas completed. We not only had many interesting lessons in geography while making this Atlas, but we had correlated it with English, drawing, and composition. I n addition to this we learned how to use books; how to cooperate with others; and how to make maps. We had much fun, enthusiasm, interest, and search work, and at the same time the boys and girls received a vast amount of knowledge. That the work was thoroly enjoyed was indicated by a request to make a similar Atlas of Germany.
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