Medium chain length polyhydroxyalkanoate produced from ethanol by Pseudomonas putida grown in liquid obtained from acidogenic digestion of organic municipal solid waste.

Production of medium chain length polyhydroxyalkanoate (mcl-PHA) up to about 6 g.L-1 was obtained by feeding ethanol to Pseudomonas putida growing in liquid obtained from acidogenic digestion of organic municipal solid waste. Washing the wet, heat-inactivated Pseudomonas cells at the end of the fermentation with ethanol obviated the need of drying the biomass and enabled the removal of contaminating lipids before solvent-mediated extraction of PHA. Using 'green' solvents, 90 to near 100% of the mcl-PHA was extracted and purities of 71-78% mcl-PHA were reached already by centrifugation and decantation without further filtration for biomass removal. The mcl-PHA produced in this way consists of 10-18% C8, 72-78% C10 and 8-12% C12 chains (entirely medium chain length), has a crystallinity and melting temperature of ∼13% and ∼49 °C, respectively, and is a stiff rubberlike, colourless material at room temperature.

Process simulation and life cycle assessment of converting autoclaved municipal solid waste into butanol and ethanol as transport fuels.

In 2015/2016, the total municipal solid waste (MSW) collected by local authority in the U.K. was 26 million tonnes and over 57% is still put into landfill or incinerated. MSW is a promising feedstock for bio-butanol production as it has a high lignocellulosic fibre content such as paper, wood, and food waste, about 50 wt% of a typical MSW stream. The study evaluates acetone, butanol, ethanol and hydrogen production from autoclaved municipal solid waste feedstock. Life cycle assessment is undertaken to evaluate the acetone, butanol, ethanol and hydrogen production process, considering cogeneration of heat and power from residual biogenic waste based on experimental data and process modelling. Acetone, butanol, and ethanol product yield can be achieved at 12.2 kg butanol, 1.5 kg ethanol, 5.7 kg acetone, and 0.9 kg hydrogen per tonne MSW. The product yield is relatively low compared to other lignocellulosic feedstocks primarily because of the lower hydrolysis yield (38% for glucose) achieved in this study; however, hydrolysis yields could be improved in future optimisation work. The conversion shows a net primary energy demand of -1.11 MJ/MJ liquid biofuels (butanol and ethanol) and net greenhouse gas emission of -12.57 g CO2eq/MJ liquid biofuels, achieving a greenhouse gas reduction of 115% compared to gasoline comparator.

Monascus ruber as cell factory for lactic acid production at low pH.

A Monascus ruber strain was isolated that was able to grow on mineral medium at high sugar concentrations and 175g/l lactic acid at pH 2.8. Its genome and transcriptomes were sequenced and annotated. Genes encoding lactate dehydrogenase (LDH) were introduced to accomplish lactic acid production and two genes encoding pyruvate decarboxylase (PDC) were knocked out to subdue ethanol formation. The strain preferred lactic acid to glucose as carbon source, which hampered glucose consumption and therefore also lactic acid production. Lactic acid consumption was stopped by knocking out 4 cytochrome-dependent LDH (CLDH) genes, and evolutionary engineering was used to increase the glucose consumption rate. Application of this strain in a fed-batch fermentation resulted in a maximum lactic acid titer of 190g/l at pH 3.8 and 129g/l at pH 2.8, respectively 1.7 and 2.2 times higher than reported in literature before. Yield and productivity were on par with the best strains described in literature for lactic acid production at low pH.

"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.

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

BACKGROUND: The 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. RESULTS: Batch 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 degrees 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. CONCLUSION: Efficient 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.

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 x h), which was lower than the production rate in media with glucose, xylose, or a combination of these sugars (9-11 mmol/[L x h]). The reduced hydrogen production rate suggests the presence of inhibiting components in paper sludge hydrolysate.

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.