Biomedical subjects

C Niklasson

Publications and source records attributed to C Niklasson.

12 recordsLinked to original sources

PubMed2005

Short circuiting in a denitrifying activated sludge tank.

The presence of a short circuit flow in a denitrifying activated sludge tank was identified and modelled. Tracer tests with pulse addition of lithium salt were used to investigate the hydraulics of the tank. The lithium concentration in the effluent was detected and residence time distribution (RTD) curves were generated. Hydraulic models based on completely stirred tank reactors (CSTRs) in series were generated from the RTD curves and the models were compared. The short circuit problem was successfully described using the Martin model, where the inflow is divided into two strands. Each strand was modelled as a number of CSTRs in series. At a normal flow the results of the model show that the tank has 12.8% dead volume, 85.8% main volume and 1.3% short circuiting volume. The inflow was divided into 91.9% entering the main volume and 8.1% entering the short circuiting volume. The mean velocity of the short circuiting stream was estimated to 0.4 m/s. At maximum flow the short circuiting stream was even larger and handled 24.3% of the flow. The short circuiting stream was identified in the upper part of the tank due to the position of the inlet and the outlet. The configuration of a tank including the use of baffles, the geometry of the inlet and mixer configuration should be considered carefully if short circuiting is to be avoided.

Bioreactors↗

PubMed2001

Effects of furfural on anaerobic continuous cultivation of Saccharomyces cerevisiae.

Furfural is an important inhibitor of yeast metabolism in lignocellulose-derived substrates. The effect of furfural on the physiology of Saccharomyces cerevisiae CBS 8066 was investigated using anaerobic continuous cultivations. Experiments were performed with furfural in the feed medium (up to 8.3 g/L) using three different dilution rates (0.095, 0.190, and 0.315 h(-1)). The measured concentration of furfural was low (< 0.1 g/L) at all steady states obtained. However, it was not possible to achieve a steady state at a specific conversion rate of furfural, q(f), higher than approximately 0.15 g/g.h. An increased furfural concentration in the feed caused a decrease in the steady-state glycerol yield. This agreed well with the decreased need for glycerol production as a way to regenerate NAD+, i.e., to function as a redox sink because furfural was reduced to furfuryl alcohol. Transient experiments were also performed by pulse addition of furfural directly into the fermentor. In contrast to the situation at steady-state conditions, both glycerol and furfuryl alcohol yields increased after pulse addition of furfural to the culture. Furthermore, the maximum specific conversion rate of furfural (0.6 g/g.h) in dynamic experiments was significantly higher than what was attainable in the chemostat experiments. The dynamic furfural conversion could be described by the use of a simple Michaelis-Menten-type kinetic model. Also furfural conversion under steady-state conditions could be explained by a Michaelis-Menten-type kinetic model, but with a higher affinity and a lower maximum conversion rate. This indicated the presence of an additional component with a higher affinity, but lower maximum capacity, either in the transport system or in the conversion system of furfural.

Anaerobiosis↗

PubMed2001

Continuous cultivation of dilute-acid hydrolysates to ethanol by immobilized Saccharomyces cerevisiae.

The continuous cultivation of immobilized Saccharomyces cerevisiae CBS 8066 on dilute-acid hydrolysates of forest residuals was investigated. The yeast cells were immobilized in 2-4% Ca-alginate beads. The 2% beads were not stable. However, the 3 and 4% beads were stable for at least 3 wk when an extra resource of calcium ions was available in the medium. The continuous cultivation of a dilute-acid hydrolysate by the immobilized cells at dilution rates of 0.3, 0.5, and 0.6 h(-1) resulted in 86, 83, and 79% sugar consumption, respectively, and an ethanol yield between 0.45 and 0.48 g/g. The hydrolysate was fermentable at a dilution rate of 0.1 h(-1) in a free-cell system but washed out at a dilution rate of 0.2 h(-1). The continuous cultivation of a more inhibiting hydrolysate was not successful by either free- or immobilized-cell systems even at a low dilution rate of 0.07 h(-1). However, when the hydrolysate was overlimed, it was fermentable by the immobilized cells at a dilution rate of 0.2 h(-1).

Alginates↗

PubMed2000

On-line control of fed-batch fermentation of dilute-acid hydrolyzates.

Dilute-acid hydrolyzates from lignocellulose are, to a varying degree, inhibitory to yeast. In the present work, dilute-acid hydrolyzates from spruce, birch, and forest residue, as well as synthetic model media, were fermented by Saccharomyces cerevisiae in fed-batch cultures. A control strategy based on on-line measurement of carbon dioxide evolution (CER) was used to control the substrate feed rate in a lab scale bioreactor. The control strategy was based solely on the ratio between the relative increase in CER and the relative increase in feed rate. Severely inhibiting hydrolyzates could be fermented without detoxification and the time required for fermentation of moderately inhibiting hydrolyzates was also reduced. The feed rate approached a limiting value for inhibiting media, with a corresponding pseudo steady-state value for CER. However, a slow decrease of CER with time was found for media containing high amounts of 5-hydroxymethyl furfural (HMF). The success of the control strategy is explained by the conversion of furfural and HMF by the yeast during fed-batch operation. The hydrolyzates contained between 1.4 and 5 g/l of furfural and between 2.4 and 6.5 g/l of HMF. A high conversion of furfural was obtained (between 65-95%) at the end of the feeding phase, but the conversion of HMF was considerably lower (between 12-40%).

Biomass↗

PubMed2000

Microaerobic glycerol formation in Saccharomyces cerevisiae.

The yeast Saccharomyces cerevisiae produces large amounts of glycerol as an osmoregulator during hyperosmotic stress and as a redox sink at low oxygen availability. NAD(+)-dependent glycerol-3-phosphate dehydrogenase in S. cerevisiae is present in two isoforms, coded for by two different genes, GPD1 and GPD2. Mutants for either one or both of these genes were investigated under carefully controlled static and dynamic conditions in continuous cultures at low oxygen transfer rates. Our results show that S. cerevisiae controls the production of glycerol in response to hypoxic conditions by regulating the expression of several genes. At high demand for NADH reoxidation, a strong induction was seen not only of the GPD2 gene, but also of GPP1, encoding one of the molecular forms of glycerol-3-phosphatase. Induction of the GPP1 gene appears to play a decisive role at elevated growth rates. At low demand for NADH reoxidation via glycerol formation, the GPD1, GPD2, GPP1, and GPP2 genes were all expressed at basal levels. The dynamics of the gene induction and the glycerol formation at low demand for NADH reoxidation point to an important role of the Gpd1p; deletion of the GPD1 gene strongly altered the expression patterns of the GPD2 and GPP1 genes under such conditions. Furthermore, our results indicate that GCY1 and DAK1, tentatively encoding glycerol dehydrogenase and dihydroxyacetone kinase, respectively, may be involved in the redox regulation of S. cerevisiae.

Aerobiosis↗

PubMed2000

Physiological effects of 5-hydroxymethylfurfural on Saccharomyces cerevisiae.

The physiological effects of 5-hydroxymethylfurfural (HMF) on Saccharomyces cerevisiae CBS 8066 in the presence and absence of furfural were studied. Experiments were carried out by pulse addition of HMF (2-4 g/l) as well as HMF (2 g/l) together with furfural (2 g/l) to batch cultivations of S. cerevisiae. Synthetic medium with glucose (50 g/l) as carbon and energy source was used. Addition of 4 g/l of HMF caused a decrease (approx. 32%) in the carbon dioxide evolution rate. Furthermore, the HMF was found to be taken up and converted by the yeast with a specific uptake rate of 0.14 (+/-0.03) g/g x h during both aerobic and anaerobic conditions, and the main conversion product was found to be 5-hydroxymethylfurfuryl alcohol. A previously unreported compound was found and characterized by mass spectrometry. It is suggested that the compound is formed from pyruvate and HMF in a reaction possibly catalysed by pyruvate decarboxylase. When HMF was added together with furfural, very little conversion of HMF took place until all of the furfural had been converted. Furthermore, the conversion rates of both furfural and HMF were lower than when added separately and growth was completely inhibited as long as both furfural and HMF were present in the medium.

Anaerobiosis↗

PubMed2000

Inhibition effects of furfural on aerobic batch cultivation of Saccharomyces cerevisiae growing on ethanol and/or acetic acid.

Physiological effects of furfural on Saccharomyces cerevisiae growing on ethanol (15 g.l(-1)) or acetate (20 g.l(-1)) as the carbon and energy source were investigated. Furfural (4 g.l(-1)), which was added during the exponential growth phase in batch cultures, was found to strongly inhibit cell growth on both carbon sources. No biomass formation occurred in the presence of furfural. However, furfural was in both cases converted to furfuryl alcohol and furoic acid, and growth resumed after complete conversion of furfural. During growth on ethanol, a rapid initial conversion of furfural to furfuryl alcohol was observed during the first few minutes after the addition of furfural, after which the conversion rate decreased to approximately 0.15 g.g(-1).h(-1) for the remaining conversion time. Acetaldehyde accumulated in the medium during the first few hours of conversion. Interestingly, addition of acetate after furfural addition resulted in an increased conversion rate of furfural and a higher carbon dioxide evolution rate, but no growth was observed until after complete conversion of furfural. Furfural addition to cells growing on acetate as the sole carbon source induced no formation of acetaldehyde, and the furfural conversion rate was lower than that on ethanol. The relationship between inhibition effects of furfural and NADH consumption is discussed.

Journal Article↗

PubMed1999

Conversion of furfural in aerobic and anaerobic batch fermentation of glucose by Saccharomyces cerevisiae.

The effect of furfural on aerobic and anaerobic batch cultures of Saccharomyces cerevisiae CBS 8066 growing on glucose was investigated. Furfural was found to decrease both the specific growth rate and ethanol production rate after pulse additions in both anaerobic and aerobic batch cultures. The specific growth rate remained low until the furfural had been completely consumed, and then increased somewhat, but not to the initial value. The CO(2) evolution rate decreased to about 35% of the value before the addition of 4 g x l(-1) furfural, in both aerobic and anaerobic fermentations. The decrease of the CO(2) evolution rate was rapid at first, and then a more gradual decrease was observed. The furfural was converted mainly to furfuryl alcohol, with a specific conversion rate of 0.6 (+/-0.03) g (furfural) x g(-1) (biomass) x h(-1) by exponentially growing cells. However, the conversion rate of furfural by cells in the stationary phase was much lower. A previously unidentified compound was detected during the conversion of furfural. This compound was characterized by mass spectrometry and it is suggested that it is formed from furfural and pyruvate.

Journal Article↗

PubMed1996

The effects of pantothenate deficiency and acetate addition on anaerobic batch fermentation of glucose by Saccharomyces cerevisiae.

Physiological effects of deficiency of pantothenate, a necessary precursor in the synthesis of coenzyme A, were studied using the yeast strain Saccharomyces cerevisiae CBS 8066. Cells were grown on defined media in anaerobic batch cultures with glucose (50 g/l) as the carbon and energy source. Batch cultures containing more than 60 micrograms/l pantothenate showed no significant differences with respect to growth rates and product yields. However, with an initial pantothenate concentration at 30 micrograms/l, the average glucose consumption rate was 50% lower than in rich medium and, at even lower concentration of pantothenate, the culture did not consume all the glucose in the medium. Furthermore, pantothenate deficiency caused the acetate and pyruvate yields to increase and the biomass yield to decrease, compared to the yields in pantothenate-rich medium. The increased acetate formation could be counteracted by initial addition of acetate to the medium, and thereby the glycerol yield could be decreased. An initial addition of acetate of 1.6 g/l to pantothenate-deficient medium (30 micrograms/l) caused a 35% decrease in glycerol yield and a 6% increase in ethanol yield. Furthermore, the time required for complete conversion of the glucose decreased by 40%. Acetate addition affected the acetate and glycerol yields in a similar way in pantothenate-rich medium (1000 micrograms/l) also.

Acetates↗

PubMed1996

Influence of the nitrogen source on Saccharomyces cerevisiae anaerobic growth and product formation.

To prevent the loss of raw material in ethanol production by anaerobic yeast cultures, glycerol formation has to be reduced. In theory, this may be done by providing the yeast with amino acids, since the de novo cell synthesis of amino acids from glucose and ammonia gives rise to a surplus of NADH, which has to be reoxidized by the formation of glycerol. An industrial strain of Saccharomyces cerevisiae was cultivated in batch cultures with different nitrogen sources, i.e., ammonium salt, glutamic acid, and a mixture of amino acids, with 20 g of glucose per liter as the carbon and energy source. The effects of the nitrogen source on metabolite formation, growth, and cell composition were measured. The glycerol yields obtained with glutamic acid (0.17 mol/mol of glucose) or with the mixture of amino acids (0.10 mol/mol) as a nitrogen source were clearly lower than those for ammonium-grown cultures (0.21 mol/mol). In addition, the ethanol yield increased for growth on both glutamic acid (by 9%) and the mixture of amino acids (by 14%). Glutamic acid has a large influence on the formation of products; the production of, for example, alpha-ketoglutaric acid, succinic acid, and acetic acid, increased compared with their production with the other nitrogen sources. Cultures grown on amino acids have a higher specific growth rate (0.52 h-1) than cultures of both ammonium-grown (0.45 h-1) and glutamic acid-grown (0.33 h-1) cells. Although the product yields differed, similar compositions of the cells were attained. The NADH produced in the amino acid, RNA, and extracellular metabolite syntheses was calculated together with the corresponding glycerol formation. The lower-range values of the theoretically calculated yields of glycerol were in good agreement with the experimental yields, which may indicate that the regulation of metabolism succeeds in the most efficient balancing of the redox potential.

Amino Acids↗

PubMed1995

Energetics and product formation by Saccharomyces cerevisiae grown in anaerobic chemostats under nitrogen limitation.

Anaerobic fermentation of glucose (20 g/l) by Saccharomyces cerevisiae CBS 8066 was studied in a chemostat (dilution rate = 0.05-0.25 h-1) at different concentrations of the nitrogen source (5.00 g/l or 0.36 g/l ammonium sulphate). The ethanol yield (g ethanol produced/g glucose consumed) was found to be higher and the glycerol yield (g glycerol formed/g glucose consumed) lower during nitrogen limitation than under carbon limitation. The biomass yield on ATP (g dry weight biomass produced/mol ATP consumed), was consequently found to be lower during nitrogen-limited conditions.

Adenosine Triphosphate↗

PubMedDate not supplied

An extended model for open-ended fluorosensor probes.

Measurements on pure solutions of NADH (8-800 microM) were made at different path lengths (0-140 mm) with an open-ended fluorosensor. It was shown that a geometric factor must be included in a model to quantitatively explain the results. A mathematical expression for this factor was found, and parameter estimations were made. It was shown that the often-used implicit assumption of a completely directed emitted light is not correct. A better fit was obtained by assuming the emitted light to be uniformly distributed in all directions. However, it was necessary to consider the slight divergence of the excitation light beams in the model.

Biosensing Techniques↗