A two-stage, two-organism process for biohydrogen from glucose

H2 can potentially be produced in a two-stage biological process: the fermentation of glucose by Escherichia coli HD701 and the photofermentation of the residual medium by Rhodobacter sphaeroides O.U. 001. In a typical batch fermentation, E. coli consumed glucose and produced H2, organic end-products and biomass. Organic end-products and residual glucose were removed during subsequent photofermentation by R. sphaeroides, with associated growth and neutralization of pH. However, photoproduction of H2 did not occur during photofermentation of the residual liquor per se due to the presence of fixed nitrogen compounds. Nevertheless, this two-stage approach could be applied to dispose of sugar-containing industrial wastes, H2 being used for on-site power generation

Integrating dark and light biohydrogen production strategies: towards the hydrogen economy

Biological methods of hydrogen production are preferable to chemical methods because of the possibility to use sunlight, CO2 and organic wastes as substrates for environmentally benign conversions, under moderate conditions. By combining different microorganisms with different capabilities, the individual strengths of each may be exploited and their weaknesses overcome. Mechanisms of bio-hydrogen production are described and strategies for their integration are discussed. Dual systems can be\ud divided broadly into wholly light-driven systems (with microalgae/cyanobacteria as the 1st stage) and partially light-driven systems (with a dark, fermentative initial reaction). Review and evaluation of published data suggests that the latter type of system holds greater promise for industrial application. This is because the calculated land area required for a wholly light-driven dual system would be too large for either centralised (macro-) or decentralised(micro-) energy generation. The potential contribution to the hydrogen economy of partially light-driven dual systems is overviewed alongside that of other biofuels such as bio-methane and bio-ethanol

Biomass-supported catalysts on Desulfovibrio desulfuricans and Rhodobacter sphaeroides

A Rhodobacter sphaeroides-supported dried, ground palladium catalyst (‘‘Rs-Pd(0)’’) was compared with a Desulfovibrio desulfuricans-supported catalyst (‘‘Dd-Pd(0)’’)and with unsupported palladium metal particles made by reduction under H2 (‘‘Chem-Pd(0)’’). Cell surface-located clusters of Pd(0) nanoparticles were detected on both D. desulfuricans and R. sphaeroides but the size and location of deposits differed among comparably loaded preparations.\ud \ud These differences may underlie the observation of different activities of Dd-Pd(0) and Rs-Pd(0) when compared with respect to their ability to promote hydrogen release from hypophosphite and to catalyze chloride release from chlorinated aromatic compounds. Dd-Pd(0) was more effective in the reductive dehalogenation of polychlorinated biphenyls (PCBs), whereas Rs-Pd(0) was more effective in the initial dehalogenation of pentachlorophenol (PCP) although the rate of chloride release from PCP was comparable with both preparations after 2 h

Hydrothermal hydrolysis of starch with CO2 and detoxification of the hydrolysates with activated carbon for bio-hydrogen fermentation.

The imminent use of hydrogen as an energy vector establishes the need for sustainable production technologies based on renewable resources. Starch is an abundant renewable resource suitable for bio-hydrogen generation. It was hypothesised that starch hydrolysates from a large (250 mL) hydrothermal reactor could support bioH2 fermentation without inhibition by toxic byproducts.\ud \ud Starch was hydrolysed at high concentrations (40 200 g.L-1) in hot compressed water (HCW) with CO2 at 30 bar in a 250 mL reactor, the largest so far for polysaccharide hydrolysis, at 180 235 °C, 15 min. Hydrolysates were detoxified with activated carbon (AC) and tested in biohydrogen fermentations. The maximum yield of glucose was 548 g.kg starch 1 carbon at 200 °C. 5 hydroxymethyl furfural, the main fermentation inhibitor, was removed by AC to support 70% more hydrogen production than the untreated hydrolysates. The potential utilization of starch hydrolysates from HCW treatment for upscaled fermentations is promising

Use of Desulfovibrio and Escherichia coli Pd-nanocatalysts in reduction of Cr(VI) and hydrogenolytic dehalogenation of polychlorinated biphenyls and used transformer oil

BACKGROUND Desulfovibrio spp. biofabricate metallic nanoparticles (e.g. ‘Bio-Pd’) which catalyse the reduction of Cr(VI) to Cr(III) and dehalogenate polychlorinated biphenyls (PCBs). Desulfovibrio spp. are anaerobic and produce H2S, a potent catalyst poison, whereas Escherichia coli can be pre-grown aerobically to high density, has well defined molecular tools, and also makes catalytically-active ‘Bio-Pd’. The first aim was to compare ‘Bio-Pd’ catalysts made by Desulfovibrio spp. and E. coli using suspended and immobilised catalysts. The second aim was to evaluate the potential for Bio-Pd-mediated dehalogenation of PCBs in used transformer oils, which preclude recovery and re-use.\ud RESULTS Catalysis via Bio-PdD. desulfuricans and Bio-PdE. coli was compared at a mass loading of Pd:biomass of 1:3 via reduction of Cr(VI) in aqueous solution (immobilised catalyst) and hydrogenolytic release of Cl- from PCBs and used transformer oil (catalyst suspensions). In both cases Bio-PdD. desulfuricans outperformed Bio-Pd E. coli by ~3.5-fold, attributable to a ~3.5-fold difference in their Pd-nanoparticle surface areas determined by magnetic measurements (Bio-PdD. desulfuricans) and by chemisorption analysis (Bio-PdE. coli). Small Pd particles were confirmed on D. desulfuricans and fewer, larger ones on E. coli via electron microscopy. Bio-PdD. desulfuricans-mediated chloride release from used transformer oil (5.6 $\pm$ 0.8 $\mu$g mL-1 ) was comparable to that observed using several PCB reference materials. \ud CONCLUSIONS At a loading of 1:3 Pd: biomass Bio-PdD. desulfuricans is 3.5-fold more active than Bio-PdE. coli, attributable to the relative catalyst surface areas reflected in the smaller nanoparticle sizes of the former. This study also shows the potential of Bio-PdD. desulfuricans to remediate used transformer oil

Polyhydroxybutyrate accumulation by a Serratia sp

A strain of Serratia sp. showed intracellular electron-transparent inclusion bodies when incubated in the presence of citrate and glycerol 2-phosphate without nitrogen source following pregrowth under carbon-limitation in continuous culture. About 1.3 mmol citrate were consumed per 450 mg\ud biomass, giving a calculated yield of maximally 55% of stored material per g of biomass dry wt. The inclusion bodies were stained with Sudan Black and Nile Red (NR), suggesting a lipid material, which was confirmed as polyhydroxybutyrate (PHB) by analysis of molecular fragments by GC and by FTIR spectroscopy of isolated bio-PHB in comparison with reference material. Multi-parameter flow cytometry in conjunction with NR fluorescence, and electron microscopy, showed that not all cells contained heavy PHB bodies, suggesting the potential for increasing\ud the overall yield. The economic attractiveness is\ud enhanced by the co-production of nanoscale hydroxyapatite\ud (HA), a possible high-value precursor for bone replacement materials

Biomanufacture of nano-Pd(0) by Escherichia coli and electrochemical activity of bio-Pd(0) made at the expense of H2 and formate as electron donors

Objective: Palladised cells of Desulfovibrio desulfuricans and Shewanella oneidensis have been reported as fuel cell electrocatalysts but growth at scale may be unattractive/costly; we have evaluated the potential of using E. coli, using H2/formate for Pd-nanoparticle manufacture. Results: Using ‘bio-Pd’ made under H2 (20 wt%) cyclic voltammograms suggested electrochemical activity of bio-NPs in a native state, attributed to proton adsorption/desorption. Bio-Pd prepared using formate as the electron donor gave smaller, well separated NPs; this material showed no electrochemical properties, and hence little potential for fuel cell use using a simple preparation technique. Bio-Pd on S. oneidensis gave similar results to those obtained using E. coli. Conclusion: Bio-Pd is sufficiently conductive to make an E. coli-derived electrochemically active material on intact, unprocessed bacterial cells if prepared at the expense of H2, showing potential for fuel cell applications using a simple one-step preparation method

Increased Hydrogen Production by Genetic Engineering of Escherichia coli

Escherichia coli is capable of producing hydrogen under anaerobic growth conditions. Formate is converted to hydrogen in the fermenting cell by the formate hydrogenlyase enzyme system. The specific hydrogen yield from glucose was improved by the modification of transcriptional regulators and metabolic enzymes involved in the dissimilation of pyruvate and formate. The engineered E. coli strains ZF1 (ΔfocA; disrupted in a formate transporter gene) and ZF3 (ΔnarL; disrupted in a global transcriptional regulator gene) produced 14.9, and 14.4 µmols of hydrogen/mg of dry cell weight, respectively, compared to 9.8 µmols of hydrogen/mg of dry cell weight generated by wild-type E. coli strain W3110. The molar yield of hydrogen for strain ZF3 was 0.96 mols of hydrogen/mol of glucose, compared to 0.54 mols of hydrogen/mol of glucose for the wild-type E. coli strain. The expression of the global transcriptional regulator protein FNR at levels above natural abundance had a synergistic effect on increasing the hydrogen yield in the ΔfocA genetic background. The modification of global transcriptional regulators to modulate the expression of multiple operons required for the biosynthesis of formate hydrogenlyase represents a practical approach to improve hydrogen production