The Emi2 Protein of Saccharomyces cerevisiae is a Hexokinase Expressed under Glucose Limitation

Hexokinases catalyze glucose phosphorylation at the first step in glycolysis in eukaryotes. In the budding yeast Saccharomyces cerevisiae , three enzymes for glucose phosphorylation have long been known: Hxk1, Hxk2, and Glk1. In this study, we focus on Emi2, a previously uncharacterized hexokinase-like protein of S. cerevisiae . Our data show that the recombinant Emi2 protein (rEmi2), expressed in Escherichia coli , possesses glucose-phosphorylating activity in the presence of ATP and Mg ²⁺ . It was also found that rEmi2 phosphorylates not only glucose but also fructose, mannose and glucosamine in vitro . In addition, we examined changes in the level of endogenous Emi2 protein in S. cerevisiae in the presence or absence of glucose and a non-fermentable carbon source. We found that the expression of Emi2 protein is tightly suppressed during proliferation in high glucose, while it is strongly upregulated in response to glucose limitation and the presence of a non-fermentable carbon source. Our data suggest that the expression of the endogenous Emi2 protein in S. cerevisiae is regulated under the control of Hxk2 in response to glucose availability in the environment.     

啤酒废酵母干燥工艺设计的研究

该文设计了一套适用于年产量5万吨以上啤酒厂废酵母干燥的生产工艺，提出了设计要点，并对干燥工艺进行了适用性分析。     

APPLICATION OF THE POLYMERASE CHAIN REACTION TO THE RAPID ANALYSIS OF BREWERY YEAST STRAINS

The rapid discrimination of closely-related Saccharomyces cerevisiae strains can pose a significant problem to breweries, in particular where closely related strains are being used simultaneously to manufacture different products. In this study, two PCR approaches have been examined to assess their usefulness for the discrimination of brewery ale and lager yeast strains. PCR using arbitrary primers (RAPD PCR) was found unsuitable for such an application since the DNA profiles generated from brewery strains were generally found to be identical, due presumably to the close genetic relatedness of these yeasts. In contrast, PCR using δ sequence primers could rapidly differentiate between many ale and lager strains and characteristic profiles for these were generated. This method could also be applied directly to yeasts isolated from brewery worts or from active dried yeast preparations. Results of such analyses were available within the working day.     

MITOCHONDRIAL RELEVANCE TO YEAST FERMENTATIVE PERFORMANCE: A REVIEW

It is well known that dissolved oxygen fulfils critical roles in brewing yeast physiology and overall fermentative performance. The major and minor roles that have been identified are briefly discussed and another role, that of providing for minimal mitochondrial development and functionality, is suggested. The long accepted theory that mitochondria are irrelevant to fermentative performance is reviewed as to its basis and the evidence in support of it. However, minimal mitochondrial development is required to provide the cell with critical metabolic intermediates and components. These are identified and reviewed and finally, evidence is presented that mitochondria are critical to brewing yeast fermentative performance. The review concludes that when assessing the role of mitochondria, concern should be broader than simply for the energetic function of these organelles.     

Assignment of most genes encoding major peroxisomal polypeptides to chromosomal band V of the asporogenic yeast Candida tropicalis

The peroxisomes of the asporogenic yeast Candida tropicalis contain about 20 major polypeptides (PXPs). We have isolated a number of genes encoding them; 11 POX genes encoded independent PXPs and three POY genes were likely to encode three other PXPs. To locate these genes on the chromosomes, chromosomes of C. tropicalis were separated by pulsed-field gel electrophoresis. Eight chromosomal bands were observed over the range of 1.0 Mbp (band 1) to 2.8 Mbp (band VIII); the genome size was estimated to be about 20 Mbp. Southern blot analysis showed that ten genes were on band V, three genes were on band IV, and the other gene was on band VI. Three genes gave hybridization signals of nearly equal intensity on two different chromosomal bands: POX6A and POX8B, on bands V and VII; and POX8A, on bands IV and VI. Ribosomal RNA genes also hybridized to two bands, VI and VII. Most genes assigned to only one band hybridized to two restriction fragments produced by either NotI or SfiI endonuclease. The results suggested that C. tropicalis was diploid and that restriction sites were conserved little between homologues. The three POX genes that were found on two chromosomal bands hybridized to not more than two restriction fragments, implying that the allelic genes were present on different chromosomal bands.     

Rheological behaviour of concentrated yeast suspension

The effect of yeast cell volumetric concentration on the rheological properties of the suspensions was measured in a pipe-flow viscometer at 30°C: at low microbial concentrations the suspensions were Newtonian; however, non-Newtonian behaviour, which could be described by the power-law equation, was observed with suspensions at high microbial volumetric concentrations. At conditions of constant microbial morphology and growth rate, the results also indicated that a relationship could be developed between the power-law constants and the microbial volumetric concentration.     

Effect of Yeast Extracts Containing Propionic Acid on Bread Dough Fermentation and Bread Properties

ABSTRACT: Three commercial yeast extracts (Oxoid, Champlain, Lallemand) fermented with immobilized cells of Propionibacterium freudenreichii were added in broths and bread formulations, and their effect on growth and gas production by bakers yeast was determined. Appearance and preservation of the breads were examined. There was significantly more gas produced by the yeast when fermented yeast extracts were added to the dough. In broths, the initial growth rate was slower with media containing fermented yeast extract when compared to their unfermented equivalents. Loaves formulated with propionic acid of the fermented yeast extract contained less ethanol. Paired t-tests showed that breads formulated with fermented yeast extract were protected for a longer period against mold than those that contained non-fermented yeast extract.     

Effects of temperature, ammonium and glucose concentrations on yeast growth in a model wine system

In enology, alcoholic fermentation is a complex process involving several mechanisms. Slow and incomplete alcoholic fermentation is a chronic problem for the wine industry and factors leading to sluggish and stuck fermentations have been extensively studied and reviewed. The most studied cause of sluggish and stuck fermentation is the nitrogen content limitation. Nevertheless, other factors, such as temperature of fermentation and sugar concentration can affect the growth of yeasts. In this study we modelled the yeast growth‐cycle in wine model system as a function of temperature, sugar and ammonium concentrations; the individual effects and the interaction of these factors were analysed by means of a quadratic response surface methodology. Cell concentrations and weight loss were monitored in the whole wine fermentation process. The results of central composite design show that lower is the availability of nitrogen, higher is the cell growth rate; moreover, initial nitrogen concentration also influences survival time of Saccharomyces cerevisiae.