Effects of metabolic inhibitors on the alcoholic fermentation by several yeasts in batch or in immobilized cell systems

Summary In previous papers it was shown that the bacterium Zymomonas mobilis might be an interesting alternative for industrial alcohol production from sugar, compared to Saccharomyces bayanus. Factors that might increase the glucose to ethanol conversion efficiency and which are in favour of the bacterium, are the production of less biomass and less by-products such as glycerol, succinic acid, butanediol, acetoin, and acetic acid. In order to reduce the synthesis of biomass three metabolic inhibitors were now studied: dinitrophenol, azide and arsenate. Their effects on the alcoholic fermentation in batch and in immobilized cell system were investigated, using three yeasts: Saccharomyces bayanus, Schizosacharomyces pombe, and Saccharomyces diastaticus. It was found that dinitrophenol in 0.1 mM concentration was effective in increasing the conversion of glucose to ethanol especially with Saccharomyces bayanus while azide in 0.1 mM concentration was better with Schizosaccharomyces pombe. In immobilized systems high steady state ethanol production from 15% glucose media was obtained by inclusion into the media of dinitrophenol or azide. Arsenate had less effect at the concentration used. Arsenate had less effect at the concentrations used. As a result ethanol productivity in g·l^-1·h^-1 was increased from around 70 in the absence of inhibitor to around 74 in the presence of dinitrophenol with Saccharomyces bayanus. With Schizosaccharomyces pombe the productivity was increased from around 65 in the absence of inhibitor to around 74 in the presence of azide. The specific ethanol productivity expressed as g ethanol formed per hour and per g viable cells was increased from 0.87 to 1.37 for Schizosaccharomyces pombe and from 1.02 to 1.66 for Saccharomyces bayanus.     

Survival of Curvularia lunata var. aeria in soil

Spore suspensions from 7 day old cultures of Curvularia lunata (Wakker) Boedijn var. aeria (Batista, Lima and Vasconcelos) M.B. Ellis were added to soil samples originally devoid of Curvularia propagules. The test fungus disappeared after six weeks of inoculation irrespective of soil amendments, like the application of green manure, urea, superphosphate, or soil cropped to Sorghum (dura) or cotton seedlings.     

Intestinal helminths of some Nile fishes near Cairo, Egypt with redescriptions of Camallanus kirandensis Baylis 1928 (Nematoda) and Bothriocephalus aegyptiacus Rysavy and Moravec 1975 (Cestoda)

Of 10 species of Nile fishes collected during the summer of 1975 at Giza near Cairo, Egypt, 7 harbored 7 helminth species. Recovered material includes 3 species of nematodes: Camallanus kirandensis Baylis 1928 (new host and geographical record), Cucullanus barbi Baylis 1923, and Rhabdochona congolensis Campana-Rouget 1961 (new host record); three species of cestodes: Khawia sp. (new in Africa), Polyonchobothrium clarias (Woodland 1925) Meggitt 1930, and Bothriocephalus aegyptiacus Rysavy and Moravec 1975; and one acanthocephalan species, Acanthosentis tilapiae Baylis 1947 (new geographical record). Morphologic observations of each species are reported. C. kirandensis and B. aegyptiacus are redescribed.     

Isolation of portulaca oleracea (regla) mucilage and identification of its structure

Portulaca oleracea leaves were found to contain 0.42% of a mucilage mixture. The mucilage was fractionated into an acidic and a neutral fraction. The acidic fraction consists of galacturonic acid residues joined by α-(1→4)-linkages; 60% of these residues are present as the calcium salt, and esterified galacturonic acid residues are absent. The neutral fraction is composed of 41% of arabinose and 43% of galactose residues, besides traces of rhamnose residues.     

Polar arrest of the simian virus 40 tumor antigen-mediated replication fork movement in vitro by the tus protein-terB complex of Escherichia coli.

The effect of the tus protein-terB sequence complex of Escherichia coli on the movement of the SV40 large tumor antigen (T antigen)-mediated replication fork during SV40 DNA replication in vitro has been examined. In the monopolymerase and dipolymerase systems, the tus protein-terB complex efficiently blocked the replication fork movement in a polar fashion, as observed in prokaryotic replication systems. With crude cytosolic extracts of HeLa cells, the same polarity of fork arrest was observed, but the block of replication fork movement was inefficient. These results indicate that the structure of the prokaryotic tus protein-terB complex allows it to block replication fork movement in an orientation-dependent manner. We also show that the tus protein-terB complex blocks the 3'----5' helicase action of T antigen in a polar fashion, using substrates comprised of single-stranded M13 DNA with either a 52-base pair (bp) or 29-bp duplex containing the terB sequence. The tus protein-terB complex formed on the 52-bp duplex was less effective than the complex formed on the 29-bp duplex in blocking the helicase action of T antigen. With the 52-bp duplex substrate, T antigen movement was only partially (30%) blocked by the tus protein-terB sequence complex in the active orientation, whereas the E. coli dnaB helicase moving 5'----3' was blocked more than 90% by the complex in the active orientation. However, with the shorter 29-bp duplex substrate, the complex blocked the T antigen helicase activity about 75%, whereas the dnaB helicase activity was completely blocked. Altogether, these results suggest that the T antigen helicase activity, when coupled to DNA replication, is more susceptible to arrest by the tus protein-terB complex than the T antigen functioning as a helicase alone.     

Conversion of sugar-beet particles to ethanol by the bacteriumZymomonas mobilis in solid-state fermentation

Summary The possibility of usingZymomonas mobilis as the microorganism, in solid-state fermentation of sugar-beet particles was investigated. The major factors affecting the process were investigated and related to ethanol yield and productivity. Ethanol yield of 0.48 g/g sugar, volumetric productivity of 12 g/L h, and final ethanol concentration of 130 g/L show the good performance ofZ.mobilis in a solid-state fermentation.     

Aphid population abundance and pestiferous effect on various bean plant species

The population abundance, infestation, and harmful effects of the aphid Aphis craccivora Koch (Hemiptera: Aphididae) were studied on four bean plant species, namely the country bean (Lablab purpureus var. BARI Seem 1), the yard‐long bean (Vigna sesquipedalis var. BARI Borboti 1), the hyacinth bean (Dolichos lablab var. BARI Seem 6), and the bush bean (Phaseolus vulgaris var. BARI Jar Seem 3). Aphid abundance and infestation on the leaves, inflorescences, flowers, and pods differed significantly among the bean plant species, with P. vulgaris and V. sesquipedalis having the lowest and highest results, respectively. Aphid severity grade and the number of trichomes of the bean plant species were negatively correlated. The duration of the growth stages among the bean plant species were significantly different, with V. sesquipedalis having the shortest durations. Aphid abundance and infestation significantly affected the physical and phytochemical characteristics of the bean plant species. The highest reduction of number of leaves, flower inflorescences, and pod inflorescences per plant, and moisture and chlorophyll content in the leaves was found in L. purpureus. The results for V. sesquipedalis revealed the highest reduction in plant height, seed weight, and pH, while those of D. lablab showed the highest reduction in leaf area.     

Optimization of parameters affecting on CHO cell culture producing recombinant erythropoietin

Abstract

Several factors may affect erythropoietin (EPO) sugar structures including designing cell culture procedure, pH, concentration of additives, dissolved oxygen, and other physicochemical parameters. In this study, we investigated the influence of changes in effective parameters and compounds on the growth rate of Chinese hamster ovary cell (CHO) cells producing recombinant EPO. Cell culture was performed at different temperature, buffering conditions, and varied concentrations of additives such as pyruvic acid, insulin, GlutaMAX, and sodium butyrate. Results indicated that the optimal temperature and pH were 37?°C and 7.2, respectively. Also, optimal concentrations for pyruvic acid, butyrate, glutamate, and insulin were obtained to be 20?mM, 1?mM, 2?mM, and 40?μg/mL, respectively. Then, cell culture was performed in microcarrier-coated spinner flasks under the optimized condition. The results showed recombinant human EPO (rhEPO) production with adequate purity. Optimization of physicochemical conditions and culture media are important factors to improve the quantity and quality of protein products. This study showed that cell growth and recombinant EPO protein production significantly increased under the optimized conditions. The results of this research can also be used in scale-up to increase the efficiency of EPO production.

Abbreviations: EPO: erythropoietin; CHO cell: Chinese hamster ovary cell; rhEPO: recombinant human EPO; DMEM: modified eagle’s medium; FBS: fetal bovine serum; SDS-PAGE: sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IGF-1: insulin-like growth factor 1     

Effectiveness of host cell protein removal using depth filtration with a filter containing diatomaceous earth

The increased cell density and product titer in biomanufacturing have led to greater use of depth filtration as part of the initial clarification of cell culture fluid, either as a stand-alone unit operation or after centrifugation. Several recent studies have shown that depth filters can also reduce the concentration of smaller impurities like host cell proteins (HCP) and DNA, decreasing the burden on subsequent chromatographic operations. The objective of this study was to evaluate the HCP removal properties of the Pall PDH4 depth filter media, a model depth filter containing diatomaceous earth, cellulose fibers, and a binder. Experiments were performed with both cell culture fluid (CCF) and a series of model proteins with defined pI, molecular weight, and hydrophobicity chosen to match the range of typical HCP. The location of adsorbed (fluorescently labeled) proteins within the depth filters was determined using confocal scanning laser microscopy. Protein binding was greater for proteins that were positively charged and more hydrophobic, consistent with adsorption to the negatively charged diatomaceous earth. The lowest degree of binding was seen with proteins near their pI, which were poorly removed by this filter. These results provide new mechanistic insights into the factors governing the filter capacity and performance characteristics of depth filters containing diatomaceous earth that are widely used in the clarification of CCF.