Sugar In Medium Research Articles

Microstructural characteristics and corrosion behaviour of high-chromium cast iron alloys in sugar media

The work is aimed at using high-Cr White Irons in manufacturing equipment for sugar cane industry.

Using inert dusts to detect and assess varroa infestations in honey bee colonies

SUMMARYDetection and assessment of varroa infestations in honey bee colonies are important for successful beekeeping. We examined the use of inert dusts to dislodge mites from adult honey bees that were isolated from their nest. Six dusts (powdered sugar, fine sugar, wheat flour, talcum powder, corn starch and baking soda) were evaluated for their ability to dislodge mites from adult bees collected in jars. We obtained the highest recovery rate with powdered sugar (92.9 ± 5.5%) and talcum powder (84.0 ± 5.6%). We also examined mite survival after recovery with inert dusts and compared it to mite survival after recovery from brood. After 24 h mite survival was significantly greater when mites were recovered with corn starch, powdered sugar, and from brood (F = 22.88, d.f. = 6,35, P < 0.0001). Finally, ether and powdered sugar were compared as tools for detecting and assessing the degree of infestation. Powdered sugar did not differ from ether in detecting or assessing low (1–5 mites per sample) infestation levels (F = 2.81, d.f. = 1,49, P = 0.1). At medium (6–30 mites per sample) and high (> 30 mites per sample) infestation levels, more mites were recovered with powdered sugar (medium: F = 14.28, d.f. = 1,29, P = 0.008; high: F = 6.34, d.f. = 1,17, P = 0.023).

Performance testing of Zymomonas mobilis metabolically engineered for cofermentation of glucose, xylose, and arabinose.

IOGEN Corporation of Ottawa, Canada, has recently built a 40t/d biomass-to-ethanol demonstration plant adjacent to its enzyme production facility. It has partnered with the University of Toronto to test the C6/C5 cofermenta-tion performance characteristics of the National Renewable Energy Labora-tory's metabolically engineered Zymomonas mobilis using various biomass hydrolysates. IOGEN's feedstocks are primarily agricultural wastes such as corn stover and wheat straw. Integrated recombinant Z. mobilis strain AX101 grows on D-xylose and/or L-arabinose as the sole carbon/energy sources and ferments these pentose sugars to ethanol in high yield. Strain AX101 lacks the tetracycline resistance gene that was a common feature of other recombinant Zm constructs. Genomic integration provides reliable cofermentation performance in the absence of antibiotics, another characteristic making strain AX101 attractive for industrial cellulosic ethanol production. In this work, IOGEN's biomass hydrolysate was simulated by a pure sugar medium containing 6% (w/v) glucose, 3% xylose, and 0.35% arabinose. At a level of 3 g/L (dry solids), corn steep liquor with inorganic nitrogen (0.8 g/L of ammonium chloride or 1.2 g/L of diammonium phosphate) was a cost-effective nutritional supplement. In the absence of acetic acid, the maximum volumetric ethanol productivity of a continuous fermentation at pH 5.0 was 3.54 g/L x h. During prolonged continuous fermentation, the efficiency of sugar-to-ethanol conversion (based on total sugar load) was maintained at >85%. At a level of 0.25% (w/v) acetic acid, the productivity decreased to 1.17 g/L x h at pH 5.5. Unlike integrated, xylose-utilizing rec Zm strain C25, strain AX101 produces less lactic acid as byproduct, owing to the fact that the Escherichia coli arabinose genes are inserted into a region of the host chromosome tentatively assigned to the gene for D-lactic acid dehydrogenase. In pH-controlled batch fermentations with sugar mixtures, the order of sugar exhaustion from the medium was glucose followed by xylose and arabinose. Both the total sugar load and the sugar ratio were shown to be important determinants for efficient cofermentation. Ethanol at a level of 3% (w/v) was implicated as both inhibitory to pentose fermentation and as a potentiator of acetic acid inhibition of pentose fermentation at pH 5.5. The effect of ethanol may have been underestimated in other assessments of acetic acid sensitivity. This work underscores the importance of employing similar assay conditions in making comparative assessments of biocatalyst fermentation performance.

Comparison of fermentative capacities of industrial baking and wild-type yeasts of the species Saccharomyces cerevisiae in different sugar media.

To compare the fermentative capacity of wild and domesticated isolates of the genus Saccharomyces. The fermentative capacity of yeasts from a variety of wild and domesticated sources was tested in synthetic dough media that mimic major bread dough types. Domesticated yeast strains were found to have better maltose-utilizing capacity than wild yeast strains. The capacity to ferment sugars under high osmotic stress was randomly distributed amongst wild and baking strains of Saccharomyces. The domestication of bakers' yeast has enhanced the ability of yeasts to ferment maltose, without a similar impact on the fermentative capacity under high osmotic conditions. This study, combined with molecular studies of both wild and domesticated yeast, showed that domestication of bakers' yeast has resulted in improved maltose utilization, apparently via the duplication and mutation of the MAL genes.

Genotypic characterization of strains of commercial wine yeastsby tetrad analysis

The objective of this work was to use tetrad analysis to define the genotypes of a number of commercially available wine yeasts for a range of characteristics related to wine making. The levels of sporulation and spore viability of 13 wine yeasts were determined. Sporulation was very low in one strain and varied from low to high in the other 12 strains. Spore viability of these 12 strains varied from 0–95% and this range was comparable to a large sample of naturally-occurring wine strains. Colonies from viable spores, predominantly from 4-spored asci, from 11 strains were characterized for the ten traits: homothallism/heterothallism, fermentation of sucrose, galactose, maltose; growth on glycerol (nonfermentable); slow growth on glucose and glycerol; level of sulfide production; copper resistance; putative presence of a recessive lethal mutation (inviability of at least two spores/tetrad); yellow pigment (in colonies) on sugar media. The number of heterozygosities for these ten characteristics varied from zero to seven in 11 strains, and eight strains were genetically distinct. Another three strains, distinct from these eight strains, were identical for the ten characteristics and also equivalent for the levels of sporulation and spore viability. Although these three strains are marketed under different designations, there is a strong probability that they were derived from a common ancestral strain. The genotypic characterization of these 11 strains constitutes an important foundation for their identification and their use in breeding programs.

Viability of plant hairy roots is sustained without propagation in low sugar medium kept at ambient temperature

The effect of sucrose concentration in the medium on the growth and resumption ability to form lateral roots was investigated using the hairy roots of pak-bung and tobacco. It was found that the growth evaluated by root tip elongation of pak-bung and tobacco hairy roots was suppressed in the medium having an initial sucrose concentration of <2.5 kg/m 3, and that the resumption abilities of both the hairy roots could be preserved when the hairy roots were kept at an initial sucrose concentration of 2.5 kg/m 3 under ambient temperature conditions. The values of maintenance energy for pak-bung and tobacco hairy roots were determined to be 0.11 and 0.12 per day, respectively, from the total sugar consumption rates. Under the oligotrophic condition of the sucrose concentration of 2.5 kg/m 3, the hairy roots were considered to exist as resting cells with maintenance metabolism, and the minimum demand for the energy source to ensure survival of the cells was met because the cells hardly multiplied and sugar consumption was not significant. In addition, long-term storage of pak-bung hairy roots in the liquid medium with 2.5 kg/m 3 sucrose was performed at 25°C. It was demonstrated that the hairy roots could maintain their resumption abilities without a serious loss of viability over 600 days and that the number of budding lateral roots per unit length of the main roots remained a value of 72 roots/m after the 600-day storage.

Alternative pathways of glucose transport in Prevotella bryantii B(1)4(1).

Prevotella bryantii B(1)4 grew faster on glucose than mannose (0.70 versus 0.45 h(-1)), but these sugars were used simultaneously rather than diauxically. 2-deoxy-glucose (2DG) decreased the growth rate of cells that were provided with either glucose or mannose, but 2DG did not completely prevent growth. Cells grown on glucose or mannose transported both (14)C-glucose and (14)C-mannose, but cells grown on glucose had over three-fold higher rates of (14)C-glucose transport than cells grown on mannose. The (14)C-mannose transport rates of glucose- and mannose-grown cells were similar. Woolf-Augustinsson-Hofstee plots were not linear, and it appeared that the glucose/mannose/2DG carrier acted as a facilitated diffusion system at high substrate concentrations. When cultures were grown on nitrogen-deficient (excess sugar) medium, isolates had three-fold lower (14)C-glucose transport, but the (14)C-mannose transport did not change significantly. (14)C-glucose and (14)C-mannose transport rates could be inhibited by 2DG and either mannose or glucose, respectively. The (14)C-glucose transport of mannose-grown cells was inhibited more strongly by mannose and 2DG than those grown on glucose. Cells grown on glucose or mannose had similar ATP-dependent glucokinase activity, and 2DG was a competitive inhibitor (K(i)=0.75 mM). Thin layer chromatography indicated that cell extracts also had ATP-dependent mannose phosphorylation, but only a small amount of phosphorylated 2DG was detected. Glucose, mannose or 2DG were not phosphorylated in the presence of PEP. Based on these results, it appeared that P. bryantii B(1)4 had: (1) two mechanisms of glucose transport, a constitutive glucose/mannose/2DG carrier and an alternative glucose carrier that was regulated by glucose availability, (2) an ATP-dependent glucokinase that was competitively inhibited by 2DG but was unable to phosphorylate 2DG at a rapid rate, and (3) virtually no PEP-dependent glucose, mannose or 2DG phosphorylation activities.

Alternative pathways of glucose transport in Prevotella bryantii B 14

Prevotella bryantii B 14 grew faster on glucose than mannose (0.70 versus 0.45 h −1), but these sugars were used simultaneously rather than diauxically. 2-deoxy-glucose (2DG) decreased the growth rate of cells that were provided with either glucose or mannose, but 2DG did not completely prevent growth. Cells grown on glucose or mannose transported both 14C-glucose and 14C-mannose, but cells grown on glucose had over three-fold higher rates of 14C-glucose transport than cells grown on mannose. The 14C-mannose transport rates of glucose- and mannose-grown cells were similar. Woolf-Augustinsson-Hofstee plots were not linear, and it appeared that the glucose/mannose/2DG carrier acted as a facilitated diffusion system at high substrate concentrations. When cultures were grown on nitrogen-deficient (excess sugar) medium, isolates had three-fold lower 14C-glucose transport, but the 14C-mannose transport did not change significantly. 14C-glucose and 14C-mannose transport rates could be inhibited by 2DG and either mannose or glucose, respectively. The 14C-glucose transport of mannose-grown cells was inhibited more strongly by mannose and 2DG than those grown on glucose. Cells grown on glucose or mannose had similar ATP-dependent glucokinase activity, and 2DG was a competitive inhibitor ( K i=0.75 mM). Thin layer chromatography indicated that cell extracts also had ATP-dependent mannose phosphorylation, but only a small amount of phosphorylated 2DG was detected. Glucose, mannose or 2DG were not phosphorylated in the presence of PEP. Based on these results, it appeared that P. bryantii B 14 had: (1) two mechanisms of glucose transport, a constitutive glucose/mannose/2DG carrier and an alternative glucose carrier that was regulated by glucose availability, (2) an ATP-dependent glucokinase that was competitively inhibited by 2DG but was unable to phosphorylate 2DG at a rapid rate, and (3) virtually no PEP-dependent glucose, mannose or 2DG phosphorylation activities.

A history of research on yeasts 2: Louis Pasteur and his contemporaries, 1850-1880

A history of research on yeasts 2: Louis Pasteur and his contemporaries, 1850-1880

Enhancement of maltose utilisation by Saccharomyces cerevisiae in medium containing fermentable hexoses.

Some industrial strains of Saccharomyces cerevisiae are unable to maintain high rates of fermentation during transition from catabolism of hexoses to maltose. This phenomenon, termed 'maltose lag', presents problems for the baking, brewing and distilling industries, which rely on yeast catabolism of mixtures of hexoses and maltose. Maltose utilisation requires the presence of maltose permease and alpha-glucosidase (maltase), encoded by MAL genes. Synthesis of these is induced by maltose and repressed by glucose. One strain of baker's yeast used in this work exhibited a marked maltose lag, whereas a second strain exhibited a shorter lag during conversion from hexose to maltose metabolism. The extent of the lag was linked to the levels of maltose permease and maltase in cells at the time of inoculation into mixed sugar medium. This view is supported by results showing that pulsing yeast with maltose to induce expression of MAL genes prior to inoculation into mixed sugar medium, enhanced sugar fermentation. Maltose pulsing of yeasts could therefore be useful for enhancing some fermentations relevant to baking and other yeast industries.

Water of hydration and cross-linking in live and dead cells

Thermal analysis has been used successfully for the in vitro estimation of aging-related changes in biological materials, such as human erythrocytes and spermatozoa, human ear cartilage and porcine scleral tissue cells. Through low-temperature DSC curves we have observed that, as a cellular population ages, the glass-transition temperature of the overall cellular constituents (−31°> T g>−50°C) shifts to higher values. Conversely, the usual procedures followed in cryobiology to improve cellular survival (i.e. procedures that use rapid freezing by vitrification in a sugar medium, like sucrose, glycerol and PPG, to reduce the amount of freezable water in the live cells) shift the T g to lower temperatures. Thus, T g has been shown to be a very good marker in the control of the in vitro vitality degree of a cellular population. Statistically, the cellular life-to-death curves, both for erythrocytes and spermatozoa, can be described by a cubic model.

Culture on sugar medium enhances photosynthetic capacity and high light resistance of plantlets grown in vitro

The significance of photosynthetic photon flux (PPF) and sugar feeding for the production of plants in vitro is only poorly understood. Nicotiana tabacum L. plantlets were grown photoautotrophically and photomixotrophically (3% sucrose) at two different PPFs (60 µmol m−2 s−1 and 200 µmol m−2 s−1) to investigate the effect of these culture parameters on photosynthetic performance and growth. Photomixotrophically‐grown plantlets showed an increase in carbohydrate content, mainly in glucose and fructose. Plant growth, dry matter accumulation and total leaf area were higher under photomixotrophic than photoautotrophic conditions. Not only biomass formation but also photosynthesis was positively affected by exogenous sucrose; the chlorophyll (Chl) content and the light‐saturated rate of photosynthetic oxygen evolution were higher in photomixotrophic plantlets. Photoinhibition occurred in plantlets that were grown photoautotrophically at the higher PPF. It became apparent as a loss in Chl content and photochemical efficiency. Photoinhibited plantlets showed a decrease in the D2/LHCII and CP47/LHCII ratios, suggesting a preferential loss of proteins from the photosystem II (PSII) core. The increased content of xanthophyll cycle pigments in photoinhibited plantlets indicated that also protective mechanisms were activated. Photomixotrophic growth of the plantlets prevented the occurrence of photoinhibitory symptoms. Therefore, we conclude that culture on sugar medium increases not only the photosynthetic potential but also the high light resistance of plantlets grown in vitro.

Effect of pyruvate on lens myo-inositol transport and polyol formation in diabetic cataract.

In diabetic cataract, sorbitol pathway flux perturbs intracellular metabolism by two putative mechanisms. The osmolyte hypothesis implicates the aldose reductase enzyme, increased rate of reduction of glucose of sorbitol and reciprocal osmoregulatory depletion of organic osmolytes (myo-inositol). Redox hypothesis favors alterations in the ratios (NADP+/NADPH and/or NADH/NAD+ as the primary cause of glucose-induced aldose reductase related defects. Increase in NADH/NAD+ promotes increased oxidation of sorbitol to fructose by polyol dehydrogenase; potential normalization of this ratio by coadministration of pyruvate (which reoxidizes NADH to NAD+ via lactate dehydrogenases reaction) was investigated. Effects of exogenous pyruvate on lens polyol formation and sodium-dependent myo-inositol (MI) cotransporter using two in vitro models of sugar cataract were determined. Rat lenses were incubated for 16 h in either normal (5.5 mM) or high sugar medium, 35.5 mM glucose or 30 mM galactose. Then lens MI influx was compared to polyol, MI and fructose content. Pyruvate did not affect MI influx or sorbitol content in lenses incubated in control medium. In 35.5 mM glucose, coadministration of pyruvate maintained lens MI influx at 76% of control values vs. 43% for lenses without pyruvate. Furthermore, pyruvate treatment diminished lens sorbitol content by 50% and increased lens sugar content (myo-inositol, fructose, lactate) and media lactate levels. Lenses incubated in high galactose medium formed galactitol with a corresponding decreased MI content. Coadministration of pyruvate had no effect on either lens sugar content (galactitol, myo-inositol, fructose) or MI influx, consistent with the fact that galactitol was not metabolized to fructose. In conclusion, pyruvate did not exert a direct effect on the MI co-transporter or prevent galactitol inhibition of MI influx. Coadministration of pyruvate with high glucose altered lens metabolism and promoted reduction of pyruvate to lactate, increased fructose, decreased sorbitol, enhanced MI influx, maintained lens MI content, implicating both osmotic and redox systems.

Production of poly-b-hydroxybutyrate by Azotobacter vinelandii in a two-stage fermentation process

Poly-b-hydroxybutyrate (PHB) production in Azotobacter vinelandii UWD, a mutant that produces PHB constitutively, was suppressed by high aeration of beet molasses medium. Thus a two-stage process was designed using aeration to promote growth and suppress PHB production in the first phase, while lower aeration of raw sugar medium containing fish peptone was used to promote PHB formation in the second phase. A PHB yield of 36 g/l and productivity of > 1 g polymer l -1 .h was obtained by this approach.

Photoautotrophic Micropropagation of Triploid Melon: II. Rooting and Plantlet Growth on Sugar-free Media

Two triploid clones of melon from the same tetraploid parent were grown in vitro with and without sugar, rooted without sugar in media both in a laboratory controlled environment chamber (in vitro) and a greenhouse acclimatization unit (ex vitro), and compared for subsequent nursery growth in the greenhouse unit. The clone `(L-14 c B) × L-14' produced more shoots in both photomixotrophic (with sucrose) or photoautotrophic (sugar-free) conditions. Both genotypes were equally likely to root in sugar-free media, and `(L-14 × B) × L-14' rooted as well from either photoautotrophic and photomixotrophic shoots but `(L-14 × B) × Mainstream' rooted less frequently from photoautotrophic shoots. Seventy-six percent (76%) of the shoots were able to root photoautotrophically in vitro, whereas 47% of the ex vitro shoots were rooted. About 85% of plantlets from all treatments survived after transfer to the nursery. After growth in the greenhouse nursery, the sizes of plants (fresh and dry weight, leaf area) were the same for either clone, from either photoautotrophic or photomixotrophic shoots. Also, after growth in the nursery, plantlets that had been rooted in vitro were larger than those rooted ex vitro. Photoautotrophic rooting demonstrates a concept for integrating micropropagation and plug-type vegetable transplant production.

The growth of Escherichia coli in glucose-limited chemostat cultures: a re-examination of the kinetics

The relationship between specific growth rate (μ) and steady-state glucose concentration was investigated for Escherichia coli ML30 in carbon-limited chemostat culture. This was made possible by the development of a method for measuring reducing sugars in culture media in the μg · 1 −1-range. Cells initially cultivated in batch culture at high glucose concentrations required long-term adaptation to nutrient-limited growth conditions in the chemostat (between 100–200 volume changes at D = 0.6 h −1) until steady-state with respect to residual glucose concentration was reached; for adapted cells, however, new steady-state glucose concentrations were usually obtained within less than 10 volume changes. A statistical evaluation of different kinetic models showed that between 0.2 h −1 < D < 0.8 h −1 the three models proposed by Monod (1942), Shehata and Marr (1971), and Westerhoff et al. (1982) described the data equally well and the applicability of the different models is discussed. Depending on the model used, calculated glucose concentrations supporting half maximum growth rate ( K s) were in the range of 40–80 μg · 1 −1. The data strongly suggest that the large differences in K s constants reported in the literature (ranging from 40 μg · 1 −1 up to 99 mg · 1 −1) are due to the use of E. coli cells adapted to different degrees to nutrient-limited growth conditions. This indicates that it is probably not possible to describe the kinetic properties of a bacterium with a single set of kinetic ’constants’.

Periplasmic aggregation limits the proteolytic maturation of the Escherichia coli penicillin G amidase precursor polypeptide.

The Escherichia coli penicillin G amidase (PGA), which is a key enzyme in the production of penicillin G derivatives is generated from a precursor polypeptide by an unusual internal maturation process. We observed the accumulation of the PGA precursor polypeptide in the insoluble material recovered after sonication of recombinant E. coli JM109 cells grown at 26 degrees C. The aggregated nature of the accumulated molecules was demonstrated using detergents and chaotropic agents in solubilization assays. The periplasmic location of the aggregates was shown by trypsin-accessibility experiments performed on the spheroplast fraction. Finally, we showed that addition of sucrose or glycerol in the medium strongly reduces this periplasmic aggregation and as a consequence PGA production is substantially increased. Thus, periplasmic aggregation of the PGA precursor polypeptide limits PGA production by recombinant E. coli and this limitation can be overcome by addition in the medium of a non-metabolizable sugar, such as sucrose, or of glycerol.

Effect of ionenes on ion permeability of erythrocyte membranes

Ionophore activity of ionenes, like that of polymyxins, is shown to be potentiated by long-chain antiions. Rapid stoichiometric anion equilibration in isoosmotic buffered sugar medium revealed that the amplitude of Cl−/OH− exchange in erythrocytes induced by an ionene-detergent complex gradually drops with the decrease of both the charge density and the number of monomeric moieties in the polymeric chain. This is also paralleled by passive K+ leakage from the cells.

Mass Production of Ginseng (Panax ginseng) Embryoids on Media Containing High Concentrations of Sugar1

A lot of ginseng embryoids were produced by culturing on high concentrations of sugar media from the embryogenic tissues obtained by moderately high temperature treatment. When the sucrose concentration was 100 g/l, the number of embryoids produced were over 100 pieces per g of inoculum weight. It was about ten times of that produced by culturing on 30 g/l of sucrose. Glucose showed an effect similar to sucrose on the basis of weight percentage. However, mannitol did not show this effect. The embryoids obtained by these processes redifferentiated to normal plantlets on culturing on the medium containing 30 g/l of sucrose. The saponin components of the tissue containing embryoids showed a similar pattern to those of natural GINSENG by HPLC.

Inhibition ofZymomonas growth by carbon sources

Zymomonas mobilis growth is slowed by 250g substrate/L. The effect is less marked in media containing 20g substrate/L and 230g non-fermentable sugar/L. This suggests that high sugar media exert more than a simple osmotic effect.