High-rate ethanol production at low pH using the anaerobic granular sludge process.

In this study, we investigated the operational performance and product spectrum of glucose-fermenting anaerobic granular sludge reactor at pH 4. A selective environment for the growth of granules was implemented by the introduction of a 2 min settling phase, a hydraulic retention time of 6 h and a solid retention time of 12 ± 3 days. The fermentation products were ethanol, lactate, and volatile fatty acids (VFA) with yields of 0.55 ± 0.03, 0.15 ± 0.02, and 0.20 ± 0.04 gram chemical oxygen demand (gCOD)/gCOD glucose, respectively. The obtained product spectrum was remarkably different from the VFA-dominated product spectrum reported in a previous study when the same system was operated at higher pH (4.5-5.5). The shift in product spectrum coincided with a shift in the microbial community structure with the dominance of eukaryotic Candida tropicalis, Pichia jaroonii, and prokaryotic Lactobacillus species instead of the Clostridia species obtained at higher pH-values. The control of the microbiomes and the associated product spectra provides bioprocess engineers with the option to tailor a suitable precursor compound mixture for subsequent chain elongation fermentation or PHA biopolymer production.

Thauera aminoaromatica MZ1T Identified as a Polyhydroxyalkanoate-Producing Bacterium within a Mixed Microbial Consortium.

Polyhydroxyalkanoates (PHAs) form a highly promising class of bioplastics for the transition from fossil fuel-based plastics to bio-renewable and biodegradable plastics. Mixed microbial consortia (MMC) are known to be able to produce PHAs from organic waste streams. Knowledge of key-microbes and their characteristics in PHA-producing consortia is necessary for further process optimization and direction towards synthesis of specific types of PHAs. In this study, a PHA-producing mixed microbial consortium (MMC) from an industrial pilot plant was characterized and further enriched on acetate in a laboratory-scale selector with a working volume of 5 L, and 16S-rDNA microbiological population analysis of both the industrial pilot plant and the 5 L selector revealed that the most dominant species within the population is Thauera aminoaromatica MZ1T, a Gram-negative beta-proteobacterium belonging to the order of the Rhodocyclales. The relative abundance of this Thauera species increased from 24 to 40% after two months of enrichment in the selector-system, indicating a competitive advantage, possibly due to the storage of a reserve material such as PHA. First experiments with T. aminoaromatica MZ1T showed multiple intracellular granules when grown in pure culture on a growth medium with a C:N ratio of 10:1 and acetate as a carbon source. Nuclear magnetic resonance (NMR) analyses upon extraction of PHA from the pure culture confirmed polyhydroxybutyrate production by T. aminoaromatica MZ1T.

Simultaneous growth and poly(3-hydroxybutyrate) (PHB) accumulation in a Plasticicumulans acidivorans dominated enrichment culture.

The wide variety of organic carbon to nitrogen and phosphorous ratios that are encountered in different wastewaters has a major impact on the poly(3-hydroxybutyrate) (PHB) accumulation potential of microbial communities. In this study we investigated the influence of the substrate composition in terms of the carbon to nitrogen (C/N) or phosphorus (C/P) ratio on the PHB accumulation performance. A multi-reactor set-up was used, enabling parallel experiments using identical inoculum of an enrichment culture dominated by Plasticicumulans acidivorans. In all experiments simultaneous PHB production and growth was observed. Generally, when trace amounts of growth nutrients were present the PHB production yield on substrate remained high for at least 12 h. Interestingly, from the carbon to nutrient ratio in the substrate, the PHB wt% could be accurately predicted in the accumulations. This study demonstrates that strict uncoupling of microbial growth and PHA accumulation is not required for achieving high cellular PHA-contents. Herewith the range of wastewaters that enable a cellular PHA content of 80 % or higher for at least 12 h is expanded to C:N and C:P-ratios exceeding COD:N of 26 gCOD:gNH4-N and COD:P of 511 gCOD:gPO4-P respectively.

Lipid recovery from a vegetable oil emulsion using microbial enrichment cultures.

BACKGROUND: Many waste streams have a relatively high vegetable oil content, which is a potential resource that should be recovered. Microbial storage compound production for the recovery of lipids from lipid-water emulsions with open (unsterilized) microbial cultures was investigated in a sequencing batch reactor using a diluted vegetable oil emulsion as model substrate. RESULTS: After feeding, triacylglycerides (TAG) were accumulated intracellular by the microbial enrichment culture and subsequently used for growth in the remainder of the sequencing batch cycle. Roughly 50% of the added TAG could be recovered as intracellular lipids in this culture. The maximum lipid storage capacity of the enrichment culture was 54% on volatile suspended solids (VSS) mass basis in a separate fed-batch accumulation experiment. The microbial community was dominated by a lipolytic fungus, Trichosporon gracile, that was responsible for intracellular lipid accumulation but also a significant fraction of lipolytic and long chain fatty-acid-utilizing bacteria was present. CONCLUSION: Herewith, we demonstrate an effective strategy for enrichment of a microbial community that can accumulate significant amounts of lipids from wastewaters without the need for sterilization of substrates or equipment. Further optimization of this process will make recovery of lipids from wastewater possible.

Modeling PHA-producing microbial enrichment cultures--towards a generalized model with predictive power.

Polyhydroxyalkanoate (PHA) production from waste streams using microbial enrichment cultures is a promising option for cost price reduction of this biopolymer. For proper understanding and successful optimization of the process, a consistent mechanistic model for PHA conversion by microbial enrichment cultures is needed. However, there is still a lack of mechanistic expressions describing the dynamics of the feast-famine process. The scope of this article is to provide an overview of the current models, investigate points of improvement, and contribute concepts for creation of a generalized model with more predictive value for the feast-famine process. Based on experimental data available in literature we have proposed model improvements for (i) modeling mixed substrates uptake, (ii) growth in the feast phase, (iii) switching between feast and famine phase, (iv) PHA degradation and (v) modeling the accumulation phase. Finally, we provide an example of a simple uniform model. Herewith we aim to give an impulse to the establishment of a generalized model.

Waste to resource: Converting paper mill wastewater to bioplastic.

In this study we investigated the feasibility of producing polyhydroxyalkanoate (PHA) by microbial enrichments on paper mill wastewater. The complete process includes (1) paper mill wastewater acidogenic fermentation in a simple batch process, (2) enrichment of a PHA-producing microbial community in a selector operated in sequencing batch mode with feast-famine regime, (3) Cellular PHA content maximization of the enrichment in an accumulator in fed-batch mode. The selective pressure required to establish a PHA-producing microbial enrichment, as derived from our previous research on synthetic medium, was validated using an agro-industrial waste stream in this study. The microbial enrichment obtained could accumulate maximum up to 77% PHA of cell dry weight within 5 h, which is currently the best result obtained on real agro-industrial waste streams, especially in terms of biomass specific efficiency. Biomass in this enrichment included both Plasticicumulans acidivorans, which was the main PHA producer, and a flanking population, which exhibited limited PHA-producing capacity. The fraction of P. acidivorans in the biomass was largely dependent on the fraction of volatile fatty acids in the total soluble COD in the wastewater after acidification. Based on this observation, one simple equation was proposed for predicting the PHA storage capacity of the enrichment. Moreover, some crucial bottlenecks that may impede the successful scaling-up of the process are discussed.

Anaerobic digestion of spent bedding from deep litter piggery housing.

This paper investigates spent litter from deep litter piggery housing as a potential substrate for farm-scale anaerobic digestion. Degradability and degradation rates were evaluated under mesophilic conditions for unused, lightly soiled (used by weaner/small pigs), and heavily soiled (used by finishing/large pigs) wheat straw, barley straw, and rice husks bedding. Apparent first order hydrolysis rate coefficients varied, but were comparable across all samples analysed (<0.1 day(-1)). Spent wheat straw was generally more degradable (approximately 60%) than spent barley straw, while spent barley straw was comparable to raw straw (40-50%), but with higher hydrolysis rates, indicating better accessibility. Rice husks were relatively poorly degradable (<20%), but degradability was improved by weathering in a pig shed. Digestion of spent barley and wheat straw litter was significantly faster (approximately twice the rate) at low (8% solids) than high (14% solids) solids loading. Rice husks degradation kinetics were not significantly influenced by solids concentration. Intrinsic methanogenic activity of heavily soiled spent wheat straw and rice husks bedding was initially poor, but achieved full activity after 40-60 days, indicating that reactor operation without external inoculum may be possible with care.