Regulation of alcoholic fermentation in batch and chemostat cultures of Kluyveromyces lactis CBS 2359

Kluyveromyces lactis is an important industrial yeast, as well as a popular laboratory model. There is currently no consensus in the literature on the physiology of this yeast, in particular with respect to aerobic alcoholic fermentation (‘Crabtree effect’). This study deals with regulation of alcoholic fermentation in K. lactis CBS 2359, a proposed reference strain for molecular studies. In aerobic, glucose-limited chemostat cultures (D=0·05–0·40 h⁻¹) growth was entirely respiratory, without significant accumulation of ethanol or other metabolites. Alcoholic fermentation occurred in glucose-grown shake-flask cultures, but was absent during batch cultivation on glucose in fermenters under strictly aerobic conditions. This indicated that ethanol formation in the shake-flask cultures resulted from oxygen limitation. Indeed, when the oxygen feed to steady-state chemostat cultures (D=0·10 h⁻¹) was lowered, a mixed respirofermentative metabolism only occurred at very low dissolved oxygen concentrations (less than 1% of air saturation). The onset of respirofermentative metabolism as a result of oxygen limitation was accompanied by an increase of the levels of pyruvate decarboxylase and alcohol dehydrogenase. When aerobic, glucose-limited chemostat cultures (D=0·10 h⁻¹) were pulsed with excess glucose, ethanol production did not occur during the first 40 min after the pulse. However, a slow aerobic ethanol formation was invariably observed after this period. Since alcoholic fermentation did not occur in aerobic batch cultures this is probably a transient response, caused by an imbalanced adjustment of enzyme levels during the transition from steady-state growth at μ=0·10 h⁻¹ to growth at μ_max. It is concluded that in K. lactis, as in other Crabtree-negative yeasts, the primary environmental trigger for occurrence of alcoholic fermentation is oxygen limitation. © 1998 John Wiley & Sons, Ltd.     

Charging for ATM services

Advances in analog and digital telecommunications technology have provided us with a wide range of telecommunications networks since the invention of the analog telephony network by Bell in 1876, such as wireless and wired CATV networks, local, metropolitan, and wide area networks, and the Internet; and a range of protocols to go with these networks, such as Internet Protocol or ATM. Charging for services delivered across these networks has always been an issue of major concern to providers and users of telecommunications services. The ability to charge accurately and efficiently for a service is directly related to the potential quality, value, flexibility, and customer care available to the customer, and, equally important, provides a means of survival, growth, and profitability to the provider. ATM is becoming a significant carrier of telecommunications services because its technology can accommodate a wide variety of services such as multimedia on demand, voice services, or videoconferencing, demanded by today's rapidly changing market. While there are many schemes in use for the charging of mobile and fixed telephony, the area of Internet charging and, in particular, ATM charging is relatively uncharted to date. In this article we address the challenge of developing methods of charging for ATM services, and show solutions in the form of technical and commercial recommendations and a practical implementation of these recommendations     

Increasing Water System Robustness in the Netherlands: Potential of Cross-Sectoral Water Reuse

Water Resources Management - Water reuse has the potential to substantially reduce the demand on groundwater and surface water. This study presents a method to evaluate the potential of water reuse...     

An update on thoron exposure in Canada with simultaneous ²²²Rn and ²²⁰Rn measurements in Fredericton and Halifax

Naturally occurring isotopes of radon in indoor air are identified as the second leading cause of lung cancer after tobacco smoking. Radon-222 (radon gas) and radon-220 (thoron gas) are the most common isotopes of radon. While extensive radon surveys have been conducted, indoor thoron data are very limited. To better assess thoron exposure in Canada, radon/thoron discriminating detectors were deployed in 45 homes in Fredericton and 65 homes in Halifax for a period of 3 months. In this study, radon concentrations ranged from 16 to 1374 Bq m(-3) with a geometric mean (GM) of 82 Bq m(-3) and a geometric standard deviation (GSD) of 2.56 in Fredericton, and from 4 to 2341 Bq m(-3) with a GM of 107 Bq m(-3) and a GSD of 3.67 in Halifax. It is estimated that 18 % of Fredericton homes and 32 % of Halifax homes could have radon concentrations above the Canadian indoor radon guideline of 200 Bq m(-3). This conclusion is significantly higher than the previous estimates made 30 y ago with short-term radon measurements. Thoron concentrations were below the detection limit in 62 % of homes in both cities. Among the homes with detectable thoron concentrations, the values varied from 12 to 1977 Bq m(-3) in Fredericton and from 6 to 206 Bq m(-3) in Halifax. The GM and GSD were 86 Bq m(-3) and 3.19 for Fredericton, and 35 Bq m(-3) and 2.35 for Halifax, respectively. On the basis of these results, together with previous measurements in Ottawa, Winnipeg and the Mont-Laurier region of Quebec, it is estimated that thoron contributes ～8 % of the radiation dose due to indoor radon exposure in Canada.     

Interplay of different NOM fouling mechanisms during ultrafiltration for drinking water production

Ultrafiltration is an emerging technology for drinking water production, but a main challenge remains the lack of understanding about fouling. This paper investigates the impact of molecular interactions between different natural organic matter (NOM) compounds on ultrafiltration fouling mechanisms. We performed dead-end filtration experiments with individual and mixed humic acid and alginate (polysaccharide). Alginate showed detrimental, but mostly reversible, flux decline and high solute retention. Our results indicate that this was caused by pore blocking transformed into cake building and weak molecular foulant-membrane and foulant-foulant interactions. In the presence of calcium, aggravated fouling was observed, related to complexation of alginate and its subsequently induced gel formation. With humic acid, more severe irreversible fouling occurred due to humic acid adsorption. Minor adsorption of alginate onto the membrane was also observed, which probably caused the substantial irreversible flux decline. The fouling characteristics in the mixtures reflected a combination of the individual humic acid and alginate experiments and we conclude, that the individual fouling mechanisms mutually influence each other. A model elucidates this interplay of the individual fouling mechanisms via hydrophobic and electrostatic interactions. In our study such an interplay resulted in an alginate cake, or gel in the presence of calcium, which is relatively irreversibly adsorbed onto the membrane by humic acid associations. This study shows the importance of mutual influences between various foulants for improved understanding of fouling phenomena. Furthermore it shows that substances with a minor individual influence might have a large impact in mixed systems such as natural water.     

Stabilization of flux during dead-end ultra-low pressure ultrafiltration

Gravity driven ultrafiltration was operated in dead-end mode without any flushing or cleaning. In contrary to general expectations, the flux value stabilized after about one week of operation and remained constant during an extended period of time (several months). Different surface water types and diluted wastewater were used as feed water and, depending on the feed water composition, stable flux values were in the range of 4-10 L h⁻¹ m⁻². When sodium azide was added to the feed water to diminish the biological activity, no stabilization of flux occurred, indicating that biological processes play an important role in the flux stabilization process. Confocal laser scanning microscopy revealed the presence of a biofouling layer, of which the structure changed over time, leading to relatively heterogeneous structures. It is assumed that the stabilization of flux is related to the development of heterogeneous structures in the fouling layer, due to biological processes in the layer. The phenomenon of flux stabilization opens interesting possibilities for application, for instance in simple and low-cost ultrafiltration systems for decentralized drinking water treatment in developing and transition countries, independent of energy supply, chemicals, or complex process control.     

Effect of specific growth rate on fermentative capacity of baker's yeast

The specific growth rate is a key control parameter in the industrial production of baker's yeast. Nevertheless, quantitative data describing its effect on fermentative capacity are not available from the literature. In this study, the effect of the specific growth rate on the physiology and fermentative capacity of an industrial Saccharomyces cerevisiae strain in aerobic, glucose-limited chemostat cultures was investigated. At specific growth rates (dilution rates, D) below 0.28 h-1, glucose metabolism was fully respiratory. Above this dilution rate, respirofermentative metabolism set in, with ethanol production rates of up to 14 mmol of ethanol . g of biomass-1 . h-1 at D = 0.40 h-1. A substantial fermentative capacity (assayed offline as ethanol production rate under anaerobic conditions) was found in cultures in which no ethanol was detectable (D < 0.28 h-1). This fermentative capacity increased with increasing dilution rates, from 10.0 mmol of ethanol . g of dry yeast biomass-1 . h-1 at D = 0.025 h-1 to 20.5 mmol of ethanol . g of dry yeast biomass-1 . h-1 at D = 0.28 h-1. At even higher dilution rates, the fermentative capacity showed only a small further increase, up to 22.0 mmol of ethanol . g of dry yeast biomass-1 . h-1 at D = 0.40 h-1. The activities of all glycolytic enzymes, pyruvate decarboxylase, and alcohol dehydrogenase were determined in cell extracts. Only the in vitro activities of pyruvate decarboxylase and phosphofructokinase showed a clear positive correlation with fermentative capacity. These enzymes are interesting targets for overexpression in attempts to improve the fermentative capacity of aerobic cultures grown at low specific growth rates.     

Regulation of alcoholic fermentation in batch and chemostat cultures of Kluyveromyces lactis CBS 2359

In order to further investigate the potential of rifamycins as antiinflammatory drugs, twenty-five semisynthetic rifamycins were tested at concentrations ranging from 10(-9) to 10(-5) M on in vitro human neutrophil functions such as locomotion, superoxide anion production, and degranulation, under different stimulatory conditions. They were also tested as antiproliferative agents on peripheral blood lymphocytes. The present semisynthetic derivatives are in general characterized by their carrying a hydrophilic substituent; they are rifamycin S or rifamycin SV derivatives carrying at C(3) either a carboxyalkyl side-chain or a glycosyl side-chain. Derivatives of the former group displayed inhibitory activities covering the whole range of activities tested, suggesting that the sum of these different effects could support their antiinflammatory activity in vivo. These derivatives, carrying a free carboxyl, are more water soluble than rifamycin SV at physiological pH, and may serve as antiinflammatory drugs for local administration, alternative to rifamycin SV, possibly giving higher efficacy and reduced side effects of pain and tissue swelling.     

Treatment of source-separated urine by a combination of bipolar electrodialysis and a gas transfer membrane.

Urine contains nutrients which can be applied usefully as a fertiliser in agriculture, but the relatively high pH can lead to ammonia evaporation. Electrodialysis with bipolar membranes was combined with an additional mass transfer unit in order to render a product containing ammonium and phosphate at a low pH. In one case, the additional mass transfer unit consisted of bubble columns placed in acid and basic concentrate streams, connected with a circulating gas phase. In the other case, the unit consisted of a gas-filled (hydrophobic) membrane placed in between the circulating acid and basic concentrate streams. The results showed that ammonia was transferred through the gas phase, but also carbonate, which is present in stored urine originating from the hydrolysis of urea. Although the pH in the product stream decreases initially, it rises above pH 7 at longer operation times. This pH increase can be attributed to a combination of proton compensating effects. The use of ammonia-selective membranes for the transfer into the acid concentrate could provide a solution to generate an ammonium phosphate product at low pH and high recoveries.