Functional characterization of Pneumocystis carinii brl1 by transspecies complementation analysis

Pneumocystis jirovecii is a fungus which causes severe opportunistic infections in immunocompromised humans. The brl1 gene of P. carinii infecting rats was identified and characterized by using bioinformatics in conjunction with functional complementation in Saccharomyces cerevisiae and Schizosaccharomyces pombe. The ectopic expression of this gene rescues null alleles of essential nuclear membrane proteins of the Brr6/Brl1 family in both yeasts.     

Application of mass and energy balance regularities to product formation

Material and energy balances for fermentation processes with product formation are developed using regularities which are frequently useful in analyzing experimental results. Available electron balances are presented and used to analyze the results of experiments in which biomass and lysine are produced in batch culture using molasses and corn extract as organic substrates. Measured values of biomass productivity, lysine productivity, O₂ consumption rate, heat evolution rate, CO₂ evolution rate, and rate of utilization of reducing sugars (RS) are used to obtain estimates of biomass energetic yield, lysine energetic yield, and fraction of substrate energy evolved as heat. For two of the three experiments, based on the entire fermentation, the measured mass rate variables are very consistent and energy measurement errors are less than 10% when proper consideration is given to both the RS and the amino acids present in the organic substrate. The average measured value of 26.5 kcal/equivalent (equiv) of available electrons transferred to oxygen compares favorably with the known average value of 27 kcal/equiv.     

SIN and the art of splitting the fission yeast cell

The septation initiation network (SIN) triggers the onset of cytokinesis in the fission yeast Schizosaccharomyces pombe by promoting contraction of the medially placed F-actin ring. SIN signaling is regulated by the polo-like kinase plo1p and by cdc2p, the initiator of mitosis, and its activation is co-ordinated with other events in mitosis to ensure that cytokinesis does not begin until chromosomes have been separated. Though the SIN controls the contractile ring, the signal originates from the poles of the mitotic spindle. Recent studies suggest that the spindle pole body may act as a dynamic assembly site for active SIN signaling complexes. In the budding yeast Saccharomyces cerevisiae the counterpart of the SIN, called the MEN, mediates both mitotic exit and cytokinesis, in part through regulating activation of the phosphoprotein phosphatase Cdc14p. Flp1p, the S. pombe ortholog of Cdc14p, is not essential for mitotic exit, but may contribute to an orderly mitosis-G1 transition by regulating the destruction of the mitotic inducer cdc25p.     

Fermentors with the power introduced by aerating gas

In the present work methods and results of investigations on optimal construction of fermentor elements by multifactor planning at a random number of levels of any factor are presented. The optimal design of column fermentor of 0.02–100 m³ volume with sieve plates containing downcomers and with power introduced by aerating gas has been worked out. Several alternative designs have been compared by examining mass transfer rates, power requirements, and other operating characteristics. Several fermentor designs with the power introduced by aerating gas are discussed with respect to their performance for cultivating various microorganisms (yeast and bacteria).     

Growth of pellets of a basidial fungus pleurotus ostreatus under various cultivation conditions

Studies on the pelletization of fungus Pleurotus ostreatus have established the pellets to differ as to the size, density and chemical composition. Changes in the pellets, taking place during culture growth, are helpful in studying the mode of their development. We have studied various conditions of cultivation, and established the factors enhancing culture growth. Prehomogenization of seeds, velocity of stirrer revolution, partial pressure of dissolved oxygen and that of other factors was established to affect the structure of pellets and parameters of culture growth.     

Biochemical reactions of Brevibacterium flavum depending on medium stirring intensity and flow structure

Authors have studied the dependence of the physiological and biochemical characteristics of lysine producers Brevibacterium flavum on medium stirring intensity upon constant pO₂ during fermentation processes. Various factors affecting the rate of biochemical reactions in cells upon a changeable intensity of medium stirring in apparata with turbine stirrers; oxygen mass transfer intensity in the system; stirring characteristics in the fermenter; biochemical changes in the cells due to gradients of energy introduced by the stirrer in the fermenter, etc. have been studied. A comparison of the experimentally established values, characterizing macrostirring (e) and microstirring (K_R), with the physiological and chemical characteristics of bacteria during fermentation made us suppose that the character of the kinetic energy distribution in the volume of fermenter is one of the factors regulating bacterial metabolism. Besides, there exists the value e_crit, upon which there can be observed essential changes of the physiological and biochemical properties of the cells. The effect of medium flows upon the cells is characterized by index F_stress, which takes into account both, the possible presence of the cells in the zone with e ≧ e_crit, and the degree of medium flow and cell interaction, numerically equal to value (e – e_crit)^0.5.     

Optimization of the analogue-sensitive Cdc2/Cdk1 mutant by in vivo selection eliminates physiological limitations to its use in cell cycle analysis

Analogue-sensitive (as) mutants of kinases are widely used to selectively inhibit a single kinase with few off-target effects. The analogue-sensitive mutant cdc2-as of fission yeast (Schizosaccharomyces pombe) is a powerful tool to study the cell cycle, but the strain displays meiotic defects, and is sensitive to high and low temperature even in the absence of ATP-analogue inhibitors. This has limited the use of the strain for use in these settings. Here, we used in vivo selection for intragenic suppressor mutations of cdc2-as that restore full function in the absence of ATP-analogues. The cdc2-asM17 underwent meiosis and produced viable spores to a similar degree to the wild-type strain. The suppressor mutation also rescued the sensitivity of the cdc2-as strain to high and low temperature, genotoxins and an anti-microtubule drug. We have used cdc2-asM17 to show that Cdc2 activity is required to maintain the activity of the spindle assembly checkpoint. Furthermore, we also demonstrate that maintenance of the Shugoshin Sgo1 at meiotic centromeres does not require Cdc2 activity, whereas localization of the kinase aurora does. The modified cdc2-asM17 allele can be thus used to analyse many aspects of cell-cycle-related events in fission yeast.     

Mitochondrial DNA origins of the Latvian clefting population

Latvia has one of the highest prevalence of isolated cleft lip with or without cleft palate (CL/P) in Europe. To clarify the genetic origins of the Latvian cleft population and establish a method for genetic mapping, mitochondrial DNA variation was studied in a population affected with clefting. One-hundred and seven subjects and 351 samples from unrelated healthy volunteers representing four anthropologically, archaeologically and ethno-linguistically different regions of Latvia were selected. The case group showed a higher frequency of haplogroups U4 (p=0.02) and U5 (p=0.0003) than in non-U haplogroups. We hypothesize that U4 and U5 mtDNA haplotype carriers may also carry susceptibility genes for clefts. Future studies will take into consideration these definitions based on mtDNA haplotypes when analyzing genetic variations and their possible contribution to CL/P susceptibility.     

Equilibrium oxygen concentration in fermentation fluids

Summary The equilibrium oxygen concentration (C₀) was determined in a complex fermentation media containing sucrose, lysine, molasses, corn steep liquor, antifoam agents and biomass. In simple systems, with all the components being dissolved, C₀ represents the oxygen solubility and linearly decreases with increasing solute concentrations. In complex solutions with multi-phase structure an increase in C₀ can be detected. It suggests that C₀ consists of two components — one being oxygen solubility, the other being determined by the amount of oxygen adsorbed on the interphase and bound by macromolecules. The presence of biomass leads to a decrease in C₀.