Miriam A. W. Budde

Yields from Glucose, Xylose, and Paper Sludge Hydrolysate During Hydrogen Production by the Extreme Thermophile Caldicellulosiruptor saccharolyticus

This study addressed the utilization of an industrial waste stream, paper sludge, as a renewable cheap feedstock for the fermentative production of hydrogen by the extreme thermophile Caldicellulosiruptor saccharolyticus. Hydrogen, acetate, and lactate were produced in medium in which paper sludge hydrolysate was added as the sole carbon and energy source and in control medium with the same concentration of analytical grade glucose and xylose. The hydrogen yield was dependent on lactate formation and varied between 50 and 94% of the theoretical maximum. The carbon balance in the medium with glucose and xylose was virtually 100%. The carbon balance was not complete in the paper sludge medium because the measurement of biomass was impaired owing to interfering components in the paper sludge hydrolysate. Nevertheless, >85% of the carbon could be accounted for in the products acetate and lactate. The maximal volumetric hydrogen production rate was 5 to 6 mmol/(L·h), which was lower than the production rate in media with glucose, xylose, or a combination of these sugars (9–11 mmol/[L·h]). The reduced hydrogen production rate suggests the presence of inhibiting components in paper sludge hydrolysate.

Efficient hydrogen production from the lignocellulosic energy crop Miscanthus by the extreme thermophilic bacteria Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana.

BackgroundThe production of hydrogen from biomass by fermentation is one of the routes that can contribute to a future sustainable hydrogen economy. Lignocellulosic biomass is an attractive feedstock because of its abundance, low production costs and high polysaccharide content.ResultsBatch cultures of Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana produced hydrogen, carbon dioxide and acetic acid as the main products from soluble saccharides in Miscanthus hydrolysate. The presence of fermentation inhibitors, such as furfural and 5-hydroxylmethyl furfural, in this lignocellulosic hydrolysate was avoided by the mild alkaline-pretreatment conditions at a low temperature of 75°C. Both microorganisms simultaneously and completely utilized all pentoses, hexoses and oligomeric saccharides up to a total concentration of 17 g l-1 in pH-controlled batch cultures. T. neapolitana showed a preference for glucose over xylose, which are the main sugars in the hydrolysate. Hydrogen yields of 2.9 to 3.4 mol H2 per mol of hexose, corresponding to 74 to 85% of the theoretical yield, were obtained in these batch fermentations. The yields were higher with cultures of C. saccharolyticus compared to T. neapolitana. In contrast, the rate of substrate consumption and hydrogen production was higher with T. neapolitana. At substrate concentrations exceeding 30 g l-1, sugar consumption was incomplete, and lower hydrogen yields of 2.0 to 2.4 mol per mol of consumed hexose were obtained.ConclusionEfficient hydrogen production in combination with simultaneous and complete utilization of all saccharides has been obtained during the growth of thermophilic bacteria on hydrolysate of the lignocellulosic feedstock Miscanthus. The use of thermophilic bacteria will therefore significantly contribute to the energy efficiency of a bioprocess for hydrogen production from biomass.

Fermentative hydrogen production from pretreated biomass: A comparative study

The aim of this work was to evaluate the potential of employing biomass resources from different origin as feedstocks for fermentative hydrogen production. Mild-acid pretreated and hydrolysed barley straw (BS) and corn stalk (CS), hydrolysed barley grains (BG) and corn grains (CG), and sugar beet extract (SB) were comparatively evaluated for fermentative hydrogen production. Pretreatments and/or enzymatic hydrolysis led to 27, 37, 56, 74 and 45 g soluble sugars/100 g dry BS, CS, BG, CG and SB, respectively. A rapid test was applied to evaluate the fermentability of the hydrolysates and SB extract. The thermophilic bacterium Caldicellulosiruptor saccharolyticus showed high hydrogen production on hydrolysates of mild-acid pretreated BS, hydrolysates of BG and CG, and SB extract. Mild-acid pretreated CS showed limited fermentability, which was partially due to inhibitory products released in the hydrolysates, implying the need for the employment of a milder pretreatment method. The difference in the fermentability of BS and CS is in strong contrast to the similarity of the composition of these two feedstocks. The importance of performing fermentability tests to determine the suitability of a feedstock for hydrogen production was confirmed.

Hydrogen production from paper sludge hydrolysate

The main objective of this study was to develop a system for the production of “renewable” hydrogen. Paper sludge is a solid industrial waste yielding mainly cellulose, which can be used, after hydrolysis, as a feedstock in anaerobic fermentation by (hyper)thermophilic organisms, such as Thermotoga elfii and Caldicellulosiruptor saccharolyticus. Tests on different medium compositions showed that both bacteria were able to produce hydrogen from paper sludge hydrolysate, but the amount of produced hydrogen and the requirement for other components differed. Hydrogen production by T. elfii strongly depended on the presence of yeast extract and salts. By contrast, C. saccharolyticus was less dependent on medium components but seemed to be inhibited by a component present in the sludge hydrolysate. Utilization of xylose was preferred over glucose by C. saccharolyticus.

“In situ” removal of isopropanol, butanol and ethanol from fermentation broth by gas stripping

In this study, the removal of IBE from aqueous solutions by gas stripping has been characterized. The effect of one or more components in the solution on the kinetics of the separation has been studied, both at 37°C and at 70°C. Gas stripping has been applied to batch, repeated batch and continuous cultures of Clostridium beijerinckii grown on a glucose/xylose mixed sugar substrate mimicking lignocellulosic hydrolysates, with the aim of finding optimal conditions for a stable IBE-producing culture with high productivity. An innovative repeated-batch process has been demonstrated in which the gas-stripping is performed at 70°C, resulting in a prolonged stable IBE culture.

Increase in phosphorylase activity during cold-induced sugar accumulation in potato tubers

SummaryThe accumulation of reducing sugars, sucrose and hexose phosphates in cv. Bintje and genotype KW77-2916 during storage at 2, 4, or 8°C was studied in relation to several catalytic activities. Bintje tubers accumulated sugars during storage at 2 or 4°C, whereas KW77-2916 showed reduced cold-sweetening at 2°C. The increase in glucose 6-phosphate and sucrose occurred concurrently and preceded the increase in reducing sugar concentration. Phosphorylase activity showed a strong interaction with temperature, storage duration and sugar accumulation in both genotypes. Invertase activity increased in Bintje concomitantly with the increase in reducing sugars, but this effect was less obvious in KW77-2916. The activities of other glycolytic and Krebs cycle enzymes showed no obvious correlation with sugar accumulation. It is suggested that the increase in phosphorylase activity acts as a triggering event in the sweetening of potato tubers during cold storage.

Possible involvement of fructose 1,6-bisphosphatase in cold-induced sweetening of potato tubers

SummaryIn measuring fructose 1,6-bisphosphatase (EC 3.1.3.11; FBPase) in potato tubers, we used anti PFP antibodies to ensure that the assay was specific for FBPase and that PPi:fructose 6-phosphate 1-phosphotransferase (EC 2.7.1.90: PFP) did not contribute to the production of fructose 6-phosphate (Fru-6-P). The involvement of cytosolic FBPase in cold sweetening was determined by monitoring enzyme activity during storage ofSolanum tuberosum cv. Erntestolz tubers at 2 or 8°C. In contrast to the rapid increase in sucrose and reducing sugars, the specific activity of FBPase showed no change and PFP rose slightly during storage at 2°C. Sugar concentrations and enzyme activities remained virtually unchanged during storage at 8°C. These data allow the interconversion of fructose 1.6-bisphosphate (Fru-1,6-P2) to Fru-6-P and vice versa by FBPase and PFP, respectively, in potato tubers, but do not support a causal relationship between coarse metabolic control of FBPase and cold-induced sweetening.