Characteristic Decrease of Nuclear Protein Kinase Activity in G1-Arrested Cells of Saccharomyces cerevisiae cdc28

An alkalopsychrotrophic strain, Micrococcus sp. 207, inducibly and extracellularly produced amylase and pullulanase. The main hydrolysis product from amylose, with a crude enzyme preparation, was maltotetraose. The optimum temperature for activity of the amylase was 60°C and that for pullulanase 55°C. The activities at 0 to 30°C exhibited similar activation energy values. In an optimized production medium at pH 9.7, the highest yields of these enzymes were obtained after cell growth at 18°C for 4 days. At pH 8.5, the yields of amylase and pullulanase became maximum after 3 days cultivation. With more prolonged cultivation, the yield of amylase but not that of pullulanase activity decreased. These enzymes were not produced at temperatures above 30°C. Sucrose was not effective as an inducer, but it stimulated cell growth and enhanced the enzyme productivities with soluble starch.     

Growth dynamics and transcriptional responses of a Red Sea Prochlorococcus strain to varying temperatures

Prochlorococcus play a crucial role in the ocean's biogeochemical cycling, but it remains controversial how they will respond to global warming. Here we assessed the response to temperature (22–30°C) of the growth dynamics and gene expression profiles of a Red Sea Prochlorococcus strain (RSP50) in a non-axenic culture. Both the specific growth rate (0.55–0.80 day⁻¹) and cell size (0.04–0.07 μm³) of Prochlorococcus increased significantly with temperature. The primary production released extracellularly ranged from 20% to 34%, with humic-like fluorescent compounds increasing up to fivefold as Prochlorococcus reached its maximum abundance. At 30°C, genes involved in carbon fixation such as CsoS2 and CsoS3 and photosynthetic electron transport including PTOX were downregulated, suggesting a cellular homeostasis and energy saving mechanism response. In contrast, PTOX was found upregulated at 22°C and 24°C. Similar results were found for transaldolase, related to carbon metabolism, and citrate synthase, an important enzyme in the TCA cycle. Our data suggest that in spite of the currently warm temperatures of the Red Sea, Prochlorococcus can modulate its gene expression profiles to permit growth at temperatures lower than its optimum temperature (28°C) but is unable to cope with temperatures exceeding 30°C.     

COMBINED EFFECTS OF TEMPERATURE AND IRON ON THE GROWTH AND PHYSIOLOGY OF THE MARINE DIATOM PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE)

Phaeodactylum tricornutum Bohlin (Bacillariophyceae) was maintained in exponential growth under Fe‐replete and stressed conditions over a range of temperatures from 5 to 30° C. The maximum growth rate (GR) was observed at 20° C (optimal temperature) for Fe‐replete and ‐stressed cells. There was a gradual decrease in the GR decreasing temperatures below the optimum temperature; however, the growth rate dropped sharply as temperature increased above the optimum temperature. Fe‐stressed cells grew at half the growth rate of Fe‐replete cells at 20° C, whereas this difference became larger at lower temperatures. The change in metabolic activities showed a similar pattern to the change in growth rate temperature aside from their optimum temperature. Nitrate reductase activity (NRA) and respiratory electron transport system activity (ETS) per cell were maximal between 15 and 20° C, whereas cell‐specific photosynthetic rate (P_cell) was maximal at 20° C for Fe‐replete cells. These metabolic activities were influenced by Fe deficiency, which is consistent with the theoretical prediction that these activities should have an Fe dependency. The degree of influence of Fe deficiency, however, was different for the four metabolic activities studied: NRA > P_cell > ETS = GR. NRA in Fe‐stressed cells was only 10% of that in Fe‐replete cells at the same temperature. These results suggest that cells would have different Fe requirements for each metabolic pathway or that the priority of Fe supply to each metabolic reaction is related to Fe nutrition. In contrast, the order of influence of decreasing the temperature from the optimum temperature was ETS > P_cell > NRA > GR. For NRA, the observed temperature dependency could not be accounted for by the temperature dependency of the enzyme reaction rate itself that was almost constant with temperature, suggesting that production of the enzyme would be temperature dependent. For ETS, both the enzyme reactivity and the amount of enzyme accounted for the dependency. This is the first report to demonstrate the combined effects of Fe and temperature on three important metabolic activities (NRA, P_cell, and ETS) and to determine which activity is affected the most by a shortage of Fe. Cellular composition was also influenced by Fe deficiency, showing lower chl a content in the Fe‐stressed cells. Chl a per cell volume decreased by 30% as temperature decreased from 20 to 10° C under Fe‐replete conditions, but chl a decreased by 50% from Fe‐replete to Fe‐stressed conditions.     

The Influence of n-Propanol on the Growth and Conidiation of Pestalotia rhododendri

Pestalotia rhododendri was exposed to vapours from 1 ml propanol solution in water and linear growth, formation of aerial hyphae and production of conidia were determined. A special Petri dish technique was used and maximum stimulation of conidial formation was induced by the vapours from a propanol concentration of 3–4 % (v/v) at 25°C. When propanol was added directly to the medium, a concentration of 1.2 × 10^?2M was optimal for growth and sporulation at 30°C. Sporulation stimulated by propanol was observed at temperatures from 20–32°C, with an optimum at 30°C. Certain observations indicated that an exposure to propanol for 24 hours was enough to induce a stimulated spore production. The stimulation was noticed on different media at 25°C, and was more pronounced at 30°C. One exception was observed. Propanol did not promote sporulation when the fungus was grown on maltagar at 30°C. Propanol 3 ° (v/v) in combination with the standard medium containing (NH₄)₂-tartrate as sole nitrogen source, inhibited the linear growth at 15–20°C, was inactive at 22.5° and 25°C, and stimulated growth at 27.5–31°C. The stimulatory effect was maximal at 30°C. Other media were tested at 25° and 30°C. At both temperatures stimulations of linear growth caused by propanol were observed with a medium containing KNO₃ as sole nitrogen source, and inhibitions with maltagar and another medium containing l -asparaginc as sole nitrogen source. The linear growth could be either inhibited or stimulated while the sporulation was stimulated.     

The green alga Dictyosphaerium chlorelloides biomass and polysaccharides production determined using cultivation in crossed gradients of temperature and light

The green microalga Dictyosphaerium chlorelloides was identified as promising microorganism for biotechnological production of exopolysaccharides (EPS). In stationary phase the culture suspension solidifies to thick gel, with very high viscosity and high content of EPS which may be interesting for many biotechnological applications. To develop cultivation protocol for maximum biomass/polysaccharide production, the optimum conditions for growth and polysaccharides production were determined in this study using the crossed gradient cultivation method. Temperature and irradiance requirements of Dictyosphaerium chlorelloides were evaluated by statistical analyses for growth rate/biomass, extracellular (EPS) and intracellular (IPS) polysaccharides contents in crossed gradients of temperature (4–45°C) and irradiance (2–18 W/m², 9.1 – 82.3 μmol/(m² s)). The maximum relative growth rate was observed at temperatures around 19.2°C and relatively low irradiances in range 2.6–11 W/m² (11.9–50.3 μmol/(m² s)). The maximum IPS production was observed at temperatures around 19.2°C and irradiance around 11 W/m² (50.3 μmol/(m² s)). The maximum production of EPS was observed at temperatures around 25.7°C and similar irradiances as IPS production. Due to temperature separation of growth and EPS production, development of cultivation protocol based controlled temperature manipulation is possible.     

Nutrient Requirement and Growth of a Synechococcus Species Isolated from Lake Balaton

A pico sized Synechococcus species isolated from Lake Balaton was studied in batch and continuous cultures. This picocyanobacterium had a pH optimum at 8.5 and a temperature optimum at 28-30°C. The I_k value for growth was 52 μEinstein m⁻² S⁻¹, the maximum growth rate 2.27 d⁻¹, the half saturation Constant of growth 1.2 μg PO₄-P I⁻¹ and the minimal cell quota 1.74 nig P g dry weight⁻¹. The dry weight of cells showed a minimum, the chlorophyll-a/biomass ratio a maximum as a function of growth rate. Above the quota of 3.4 fg P Cell⁻¹ significant amounts of non-reactive dissolved Phosphorous were released.     

Effects of Supraoptimal Temperatures on Merulius lacrymans

The growth rate of Merulius lacrymans (Jacq.) Fr. decreases very rapidly at temperatures above optimum (about 22°C). No growth occurred above 28°C. The thermal death-time of the fungus at different temperatures above maximum was determined. At 37.5°C the thermal death-time is 4 hours. At this temperature, the respiration is not directly affected, since an appreciable respiration occurred even after 6 hours. On the basis of ¹⁴C-experiments and analysis of UV-absorbing material excreted from the mycelium at supraoptimal temperatures, it is suggested that the detrimental effects of supraoptimal temperatures on this fungus include a degradation of the nucleic acids and a subsequent leakage of the nucleotides out of the cells.     

Temperature effects on ethanol and isopropanol utilization by Candida krusei

Candida Krusei has a optimum growth temperature of 37°C on SASOL ethanol-isopropanol mixture. The organism was unable to grow on isopropanol, but oxidized it partially to acetone in the presence and absence of ethanol. Growth at 40°C in the alcohol mixture was slightly faster than at 30°C over an ethanol concentration range of 0.43 to 3.6% (v/v), although at both temperatures the growth rate declined continuously with increasing concentration. At an ethanol concentration greater than 3.6% (v/v), the mixture was much more inhibitory to growth at 40 and 30°C. The inhibitory effect was due to the ethanol rather than the isopropanol. Metabolites such as acetate, acetaldehyde, and ethyl acetate accumulated in the medium, but the degree of accumulation depended upon the temperature and alcohol mixture concentration. At 40°C, acetaldehyde and acetate accumulated to a greater extent than 30°C on a 4.0% (v/v) synthetic alcohol mixture and this may also cause the greater inhibition at this temperature. The alcohol mixture is unsuitable for single cell protein (SCP) production in batch culture because of the low cell densities observed at all alcohol concentrations.     

Effect of culture temperature on follicle-stimulating hormone production by Chinese hamster ovary cells in a perfusion bioreactor

Follicle-stimulating hormone (FSH) was produced in Chinese hamster ovary (CHO) cells using a perfusion bioreactor. Perfusion culture at 37°C yielded a high cell density but a low FSH production. To investigate the effect of culture temperature in the range of 26–37°C on cell growth and FSH production, batch cultures were performed. Lowering culture temperature below 32°C resulted in growth suppression. However, specific productivity of FSH, q _FSH, increased as culture temperature decreased, and the maximum q _FSH of 43.4 ng/10⁶ cells/h was obtained at 28°C, which is 13-fold higher than that at 37°C. Based on the results obtained from batch cultures, we performed perfusion cultures with two consecutive temperatures. CHO cells were grown up to 3.2 × 10⁷ cells/ml at 37°C and culture temperature shifted down to 28°C to obtain a high FSH titer. Soon after the maximum FSH titer of 21 μg/ml was achieved, a rapid loss of not only viable cell concentration but also cell viability was observed, probably due to the low activities of enzymes related to cell growth. Thus, the extension of production period at 28°C is critical for the enhancement of FSH production, and the use of antiapoptotic genes seems to be promising.     

General introduction to the lists of threatened biotopes,flora and fauna of the trilateral Wadden Sea Area (red data book)

The effect of the acclimation temperature on the temperature tolerance ofPorphyra leucosticta, and on the temperature requirements for growth and survival ofEnteromorpha linza was determined under laboratory conditions. Thalli ofP. leucosticta (blade or Conchocelis phases), acclimated to twenty-five degrees, survived up to 30°C, i.e. 2°C more than those acclimated to 15°C which survived up to 28°C. Lower temperature tolerance of bothPorphyra phases that were acclimated to 15°C was −1°C after an 8-week exposure time at the experimental temperatures. The upper temperature tolerance ofE. linza also increased by 2°C, i.e. from 31 to 33°C, when it was acclimated to 30°C instead of 15°C. The lower temperature tolerance increased from 1 to −1°C, when it was acclimated to 5°C instead of 15°C.E. linza thalli acclimated for 4 weeks to 5 or 10°C reached their maximum growth at 15°C, i.e. at a 5°C lower temperature than those acclimated to 15 or 30°C. These thalli achieved higher growth rates in percent of maximal growth at low temperatures than those acclimated to 15 or 30°C. Thalli acclimated for 1 week to 5°C reached their maximum growth rate at 20°C and achieved growth rates at low temperatures similar to those recorded for thalli acclimated to 15°C. Thalli ofE. linza acclimated for 4 weeks to 5°C lost this acclimation after being post-cultivated for the same period at 15°C. That was not the case with thalli acclimated for 8 weeks to 5°C and post-acclimated for 4 weeks to 15°C. These thalli displayed similar growth patterns at 10–25°C, while a decline of growth rate was observed at 5 or 30°C. The significance of the acclimation potential ofE. linza with regard to its seasonality in the Gulf of Thessaloniki, and its distribution in the N Atlantic, is also discussed.     

The Influence of <Emphasis Type="Italic">P. fluorescens</Emphasis> Cell Morphology on the Lytic Performance and Production of Phage ϕIBB-PF7A

This study aims at assessing the influence of Pseudomonas fluorescence cell morphology on the effectiveness and production of the lytic bacteriophage ϕIBB-PF7A. P. fluorescens were cultured as rods or as elongated cells by varying the temperature and rotary agitation conditions. Cells presented rod shape when grown at temperatures up to 25°C and also at 30°C under static conditions, and elongated morphology only at 30°C when cultures were grown under agitation. Elongated cells were 0.4 up to 27.9 μm longer than rod cells. Rod-shaped hosts were best infected by phages at 25°C which resulted in an 82% cell density reduction. Phage infection of elongated cells was successful, and the cell density reductions achieved was statistically similar (P > 0.05) to those obtained at the optimum growth temperature of P. fluorescens. Phage burst size varied with the cell growth conditions and was approximately 58 and 153 PFU per infected rod and elongated cells, grown at 160 rpm, at 25°C (the optimal temperature) and 30°C, respectively. Phage adsorption was faster to elongated cells, most likely due to the longer length of the host. The surface composition of rod and elongated cells is similar in terms of outer membrane proteins and lipopolysaccharide profiles. The results of this study suggest that the change of rod cells to an elongated morphology does not prevent cells from being attacked by phages and also does not impair the phage infection.     

Production of L-malate from fumarate by the yeast Dipodascus magnusii

An alternative microbiological method for the production of malate from fumarate is presented. The yeast Dipodascus magnusii was used for this bioconversion. The optimum cell growth temperature was 28°C and the working volume 120 ml. The highest level of fumarase activity during bioconversion was achieved at a pH of 7.5 and a temperature of 37°C. These conditions were determined as optimal. Using sodium fumarate (1M), the maximum specific productivity of malic acid obtained was 1.72 g/(g_DCW × h) for intact cells. In the case of ammonium fumarate, it was 2.25 g/(g_DCW × h).     

Ecophysiological adaptations of two Mediterranean red algae in relation to distribution

Effects of irradiance and temperature on the Mediterranean red algae Eupogodon spinellus and Eupogodon planus were tested. Growth of both species was saturated at an irradiance of 10–20?μmol?m^?2?s^?1, which is in accordance with their sublittoral habitat. Eupogodon spinellus and E. planus survived permanently at temperatures between 8 and 30?°C. The temperature optimum for growth was 25?°C with suboptimal growth occurring at (10?)15 and 30?°C in both species. At their collection locality (Corsica), potential monthly growth yields would be highest in summer and in winter would be only about 20% of the maximum. Reproductive requirements could be determined only in E. planus. Gametophytes reproduced both in long and in short days but only at 20?°C. Tetrasporophytes reproduced at 15–20?°C but only in short days. Geographic distribution boundaries are not set by growth or survival limits. However, the reproductive requirements of E. planus did account for its restricted distribution in the Mediterranean and on the Canary Islands.     

Direct ethanol production from cellulosic materials at high temperature using the thermotolerant yeast Kluyveromyces marxianus displaying cellulolytic enzymes

To exploit cellulosic materials for fuel ethanol production, a microorganism capable of high temperature and simultaneous saccharification–fermentation has been required. However, a major drawback is the optimum temperature for the saccharification and fermentation. Most ethanol-fermenting microbes have an optimum temperature for ethanol fermentation ranging between 28 °C and 37 °C, while the activity of cellulolytic enzymes is highest at around 50 °C and significantly decreases with a decrease in temperature. Therefore, in the present study, a thermotolerant yeast, Kluyveromyces marxianus, which has high growth and fermentation at elevated temperatures, was used as a producer of ethanol from cellulose. The strain was genetically engineered to display Trichoderma reesei endoglucanase and Aspergillus aculeatus β-glucosidase on the cell surface, which successfully converts a cellulosic β-glucan to ethanol directly at 48 °C with a yield of 4.24 g/l from 10 g/l within 12 h. The yield (in grams of ethanol produced per gram of β-glucan consumed) was 0.47 g/g, which corresponds to 92.2% of the theoretical yield. This indicates that high-temperature cellulose fermentation to ethanol can be efficiently accomplished using a recombinant K. marxianus strain displaying thermostable cellulolytic enzymes on the cell surface.     

Germination behaviour of Conyza bonariensis to constant and alternating temperatures across different populations

Conyza bonariensis is one of the most problematic weed species throughout the world. It is considered highly noxious due to its interference with human activities, and especially the competition it poses with economically important crops. This research investigated the temperature requirements for seed germination of four populations of C. bonariensis with distinct origin and the influence of daily alternating temperatures. For this, a set of germination tests were performed in growth chambers to explore the effect of constant and alternating temperatures. Seeds of the four populations (from Lleida, Badajoz and Seville, Spain and Bahía Blanca, Argentina) were maintained at constant temperatures ranging from 5 to 35°C. The final germination and cardinal temperatures (base, optimum and maximum) of each population were obtained. We also tested the influence of daily alternating temperatures on final germination. To do so, seeds were exposed to two temperature regimes: 5/15, 10/20, 15/25, 20/30 and 25/35°C night/day temperature (intervals increasing 5°C, with constant oscillation of 10°C) and to 18/22, 16/24, 14/26, 12/28 and 10/30°C night/day temperature (intervals with average of 20°C, but increasing the oscillation in 4°C between intervals). In general, all populations behaved similarly, with the highest germination percentages occurring in the optimum temperature range (between 21.7°C and 22.3°C) for both constant and alternating temperatures. In general, climatic origin affected germination response, where seeds obtained from the coldest origin exhibited the highest germination percentage at the lowest temperature assayed. In addition, we observed that the alternating temperatures can positively affect total germination, especially in oscillations that were further from the average optimum temperature (20°C), with high germination percentage for the oscillations of 15/25, 20/30, 18/22, 16/24, 14/26, 12/28 and 10/30°C in all populations. The cardinal temperatures obtained were significantly different across the populations. These results provide information that will facilitate a better understanding of the behaviour of Conyza and improve current field emergence models.     

Maintaining mosquito cell lines at high tempetatures: Effects on the replication of flaviviruses

Summary The upper thermal limit for maintenance of eleven mosquito cell lines was studied. Although most cell lines could be grown at 32°C to 34°C,Anopheles stephensi cell line could be maintained at 37°C. At higher temperatures initial growth rate was higher, but yield of cells after about a week of incubation was lower than at the standard temperature (28°C). Replication of several flaviviruses inAedes albopictus cell cultures adapted to 34.5°C was faster, and viral titers were higher than at 28°C.     

Effect of temperature on ethanol tolerance of thermotolerant Isshatchenkia orientalis IPE100

Tolerance to high temperature and ethanol is a major factor in high‐temperature bio‐ethanol fermentation. The inhibitory effect of exogenously added ethanol (0–100 g L^?1) on the growth of the newly isolated thermotolerant Issatchenkia orientalis IPE100 was evaluated at a range of temperatures (30–45°C). A generalized Monod equation with product inhibition was used to quantify ethanol tolerance, and it correlated well with the experimental data on microbial growth inhibition of ethanol at the temperatures of 30–45°C. The maximum inhibitory concentration of ethanol for growth (P_m) and toxic power (n) at the optimal growth temperature of 42°C were estimated to be 96.7 g L^?1 and 1.23, respectively. The recently isolated thermotolerant I. orientalis IPE100 shows therefore a strong potential for the development of future high‐temperature bio‐ethanol fermentation technologies. This study provides useful insights into our understanding of the temperature‐dependent inhibitory effects of ethanol on yeast growth.     

Effect of temperature on growth parameters of psychrophilic bacteria and yeasts

Three bacterial (Pedobacter heparinus, Pedobacter piscium, Pedobacter cryoconitis) and three yeast strains (Saccharomyces cerevisiae, Leucosporidiella creatinivora, Rhodotorula glacialis) of different thermal classes (mesophiles and psychrophiles) were tested for the effect of temperature on a range of growth parameters, including optical density, viable cell numbers, and cell dry mass, in order to determine the temperature conditions under which maximum biomass formation is obtained. Maximum values of growth parameters obtained at the stationary growth phase of the strains were used for statistical calculation. Temperature had a significant (P ≤ 0.05) effect on all growth parameters for each strain; correlations between the growth parameters were significant (P ≤ 0.05–0.01). The maximum growth temperature or the temperature at which microbial growth was fastest was in no case the temperature at which the investigated strains produced the highest amount of biomass. All tested psychrophilic bacteria and yeast strains produced highest amounts of cells (as calculated per mg cell dry mass or per OD₆₀₀ unit) at 1°C, while cell numbers of mesophiles were highest at 20°C. Thus, cultivation temperatures close to the maximum growth temperature are not appropriate for studying psychrophiles.     

Growth of juvenileArctica islandica under experimental conditions

In two laboratory experiments, the effects of temperature and food availability on the growth of 10- to 23-mm high specimens of the bivalveArctica islandica were estimated. Each experimental set-up consisted of 5 treatments in which either the food supply or the temperature differed. It was demonstrated thatArctica is able to grow at temperatures as low as 1°C. A tenfold increase of shell growth was observed at temperatures between 1° and 12°C. The greatest change in growth rate took place between 1° and 6°C. Average instantaneous shell growth varies between 0.0003 at 1°C to 0.0032/day at 12°C. The results suggest that temperature hardly affects the time spent in filtration, whereas particle density strongly influences that response. Starved animals at 9°C have their siphons open during only 12% of the time, whereas the siphons of optimally fed animals were open on average during 76% of the observations. Increased siphon activity corresponded to high shell and tissue growth. At 9°C, average shell growth at the optimum cell density of 20×10⁶ cell/l was 3.1 mm corresponding to an instantaneous rate of 0.0026/day. An algal cell density (Isochrysis galbana, Dunaliella marina) ranging between 5 and 7×10⁶ cell/l is just enough to keep shells alive at 9°C. Carbon conversion efficiency at 9°C is estimated to vary between 11 and 14%.