Industrial Baker Research Articles

An industrial baker's yeast strain improved for use in frozen dough

An industrial baker's yeast strain improved for use in frozen dough

Lysine Overproduction Mutations in the YeastSaccharomyces cerevisiae Are Introduced into Industrial Yeast Strains

Yeast mutants resistant to a toxic lysine analog, thialysine were obtained by a method described in the literature [1]. A strain excreting the maximum amount of lysine (0.45 g/l) was selected from these mutants. The intracellular content of lysine was also increased by 30%. The genetic nature of lysine overproduction was studied in this strain. An increase in the amount of excreted lysine was shown to be determined by at least two genes, one of which carries a mutation of thialysine resistance manifesting the pleiotropic effect of lysine overproduction (Th1R) and the other is involved in the regulation of lysine production (PRL). Linkage groups of these genes were determined: the first gene was mapped to the IV chromosome and the second, to the XV chromosome. Both genetic characters were introduced into industrial baker's yeast strains via a series of backcrosses. The stabilization of the genome in the newly derived strains was confirmed by electrokaryotyping.

Stable high-copy-number integration of Aspergillus oryzae alpha-AMYLASE cDNA in an industrial baker's yeast strain.

The Aspergillus oryzae alpha-amylase cDNA was placed under the control of the Saccharomyces cerevisiae actin promoter (pACT1) and introduced into the ribosomal DNA locus of an industrial baker's yeast strain. To obtain a strain eligible for commercial use, we constructed an integrative cassette lacking bacterial DNA sequences but containing the alpha-amylase cDNA and ribosomal DNA sequences to target the integration to this locus. High-copy-number integrants were obtained including a defective TRP1d promoter in the integrative cassette. We selected one transformant, Rib-AMY (CECT10872), in which the multi-integrated sequences were stable even after 200 generations of growth in nonselective medium. This transformant also expressed and secreted high levels of alpha-amylase. Bread made with this strain had a higher volume, lower density, and softer crumbs than bread made with a control strain. The Rib-AMY transformant also was useful in retarding bread firming. This new strain fulfills all the requirements for commercial utilization and should reduce or eliminate the requirement for addition of exogenous alpha-amylase to the flour, reducing allergenic work-related symptoms due to this enzyme.

Stress tolerance: the key to effective strains of industrial baker's yeast.

Application of yeasts in traditional biotechnologies such as baking, brewing, distiller's fermentations, and wine making, involves them in exposure to numerous environmental stresses. These can be encountered in concert and sequentially. Yeast exhibit a complex array of stress responses when under conditions that are less than physiologically ideal. These responses involve aspects of cell sensing, signal transduction, transcriptional and posttranslational control, protein-targeting to organelles, accumulation of protectants, and activity of repair functions. The efficiency of these processes in a given yeast strain determines its robustness, and to a large extent, whether it is able to perform to necessary commercial standards in industrial processes. This article reviews aspects of stress and stress response in the context of baker's yeast manufacturing and applications, and discusses the potential for improving the general robustness of industrial baker's yeast strains, in relation to physiological and genetic manipulations.

Construction of Baker's Yeast Strains that Secrete Different Xylanolytic Enzymes and their use in Bread Making

cDNAs corresponding to theAspergillus nidulansendoxylanase X22 (xlnA), X24 (xlnB) and X34 (xlnC) and to theAspergillus nigerα-L-arabinofuranosidase (abfB) were placed under the control of theSaccharomyces cerevisiaeactin promoter (pACT1) and introduced into industrial baker's yeast. OnlyA. nidulansendoxylanase X24 andA. nigerα-L-arabinofuranosidase were expressed and secreted properly. Bread made with transformant YEpACT-X24 (containing thexlnBcDNA) showed an increase in volume around 5%. The combined action of endoxylanase X24 and α-L-arabinofuranosidase on bread did not improve the effects produced by the endoxylanase alone.

Combined Expression of Aspergillus nidulans Endoxylanase X24 and Aspergillus oryzae (alpha)-Amylase in Industrial Baker's Yeasts and Their Use in Bread Making

The Aspergillus nidulans endoxylanase X24 and the Aspergillus oryzae (alpha)-amylase cDNAs were placed under the control of the Saccharomyces cerevisiae actin promoter (pACT1) and introduced into baker's yeast. Bread made with transformants expressing both enzymes (YEpACT-AMY-ACT-X24) showed a 30% increase in volume and reduced firmness in comparison with that produced with a commercial strain. Endoxylanase X24 and (alpha)-amylase seem to act synergistically to improve the quality of bread in terms of volume and density.

Heat flux measurements for the fast monitoring of dynamic responses to glucose additions by yeasts that were subjected to different feeding regimes in continuous culture.

Heat measurements have been successfully as an analytical tool for the study of the dynamics of energy metabolism of Saccharomyces cerevisiae and Candida utilis grown in continuous culture under fluctuating substrate supply. A low average dilution rate (D = 0.05 h-1) was maintained either by adding the medium as continuously (dropwise) as possible or (blockwise) by adding the medium at high speed during a short period (D = 0.5 h-1 for 40 s) and not at all during the following period (D = 0.00 h-1 for 360 s). The resulting biological activity was monitored on-line with conventional (O2 and CO2) off-gas analyses, DOT measurements, and heat flux measurements. In C. utilis cultures, the biomass-specific maximum oxygen consumption rate (qO2,max), the biomass yield (Ys,x), and the dynamic responses to a glucose pulse and to a change in feeding regime were not significantly affected by different preceding feeding regimes. In contrast, S. cerevisiae grown in continuous culture with blockwise feed showed a 50% increase in qO2,max and a 25% drop in Ys,x compared to the culture grown with dropwise feed. The dynamic response to a glucose pulse (0.6 g L-1) was slower for the continuous (dropwise) than for the blockwise grown S. cerevisiae. With a second testing method for the dynamic response of the yeasts, the feeding regime was changed. The blockwise fed S. cerevisiae proved to be better "trained" to cope with sudden changes in glucose supply and, therefore, was more "shockproof" toward a change in feeding regime. This clearly points to major differences in the intracellular metabolic flux control between the yeasts. These findings are of relevance for industrial baker's yeast production, where reactor mixing times of one to several minutes are not uncommon. The observed, heat production, together with the dissolved oxygen concentration, appeared to give the fastest response to actual changes in the culture. It is suggested that heat measurements can be a very useful tool to monitor and control the growth of S. cerevisiae in laboratory and industrial fermenter operations.

Simulation of Profit Optimization for Industrial Baker's Yeast Continuous Fermentation

An on-line profit optimization approach for baker’s yeast continuous fermentation was developed. The profit optimization was done based on the process model system and profit function which were earlier established. An on-line rolling identification method with moving-data-windows technique was used to modify the model system so as to adapt the different transients caused by different initial conditions from run to run and the time-varied dynamics of cell metabolism. With the help of such an adaptive process model, optimal molasses feeding strategies were determined by maximizing the profit function. Another primary manipulating variable, the process water feeding rate, was found to have no significant influence on the profit optimization within its normal operation range.

Differential importance of trehalose in stress resistance in fermenting and nonfermenting Saccharomyces cerevisiae cells.

The trehalose content in laboratory and industrial baker's yeast is widely believed to be a major determinant of stress resistance. Fresh and dried baker's yeast is cultured to obtain a trehalose content of more than 10% of the dry weight. Initiation of fermentation, e.g., during dough preparation, is associated with a rapid loss of stress resistance and a rapid mobilization of trehalose. Using specific Saccharomyces cerevisiae mutants affected in trehalose metabolism, we confirm the correlation between trehalose content and stress resistance but only in the absence of fermentation. We demonstrate that both phenomena can be dissociated clearly once the cells initiate fermentation. This was accomplished both for cells with moderate trehalose levels grown under laboratory conditions and for cells with trehalose contents higher than 10% obtained under pilot-scale conditions. Retention of a high trehalose level during fermentation also does not prevent the loss of fermentation capacity during preparation of frozen doughs. Although higher trehalose levels are always correlated with higher stress resistance before the addition of fermentable sugar, our results show that the initiation of fermentation causes the disappearance of any other factor(s) required for the maintenance of stress resistance, even in the presence of a high trehalose content.

Construction of industrial baker's yeast strains able to assimilate maltose under catabolite repression conditions

Spore progeny from an industrial baker's yeast strain were mutagenized with UV and mutants resistant to 2-deoxyglucose isolated. One of these mutants (10a12–13) showed high levels of maltase (α-glucosidase) and external invertase, and assimilated maltose when growing under catabolite repression conditions. This mutant was not allelic to any of the catabolite repression mutants tested cat4, cat80, cid1, cyc8, hex2, hxk2 and tup1. Mutant 10a12–13 was crossed with appropriate strains to construct hybrids that were also able to assimilate maltose in the presence of glucose. These hybrids may be useful in fermentation processes where both glucose and maltose are present.

Construction of industrial baker's yeast strains able to assimilate maltose under catabolite repression conditions

Construction of industrial baker's yeast strains able to assimilate maltose under catabolite repression conditions

Electrofusion of an industrial Baker’s yeast strain with a sour dough yeast

A method for hybridization of yeast protoplasts ofSaccharomyces cerevisiae andCandida holmii by electrofusion was optimized. The hybrids were screened on maltose-acetate agar plates. The average fusion frequency was 1.1xl0-3. Two hybrids of 132 collected from selection plates were found to be stable over 15 sequential shake flask cultivations. However, the strains reverted during production of baker’s yeast in a laboratory scale process imitating the industrial process.

Changes in the rate of fermentation of maltose during propagation of industrial baker's yeast.

The rate of fermentation of glucose and maltose and the maltase activity of cellfree preparations of yeast were investigated during yeast propagation at the individual production stages. It was found that the yeast cells do not change much in their fermentation of glucose but that the level of the maltose-hydrolyzing enzyme undergoes changes, together with the character of the anaerobic fermentation of maltose, depending on the character of cultivation (batch or incremental feeding). After an initial decrease the maltose activity of cell-free preparations is maintained practically on the same level until the expedition phase is reached when it rapidly decreases to low values. The basis of the changes investigated is discussed together with their importance for yeast production technology.

Centralization and Decentralization in Industrial Relations, BAKER, H. and FRANCE. R., Industrial Relations Section, Princeton University, 210 pp., 1954.

Centralization and Decentralization in Industrial Relations, BAKER, H. and FRANCE. R., Industrial Relations Section, Princeton University, 210 pp., 1954.. Un article de la revue Relations industrielles / Industrial Relations (Volume 10, numéro 2, mars 1955, p. 74-142) diffusée par la plateforme Érudit.