Sustainable PHA production in integrated lignocellulose biorefineries.

In emerging bioeconomies, the compostable biopolymers polyhydroxyalkanoates (PHAs) are desirable products due to their similarity to petropolymers. While industrial PHA production has been growing rapidly, obtaining a cheap and sustainable carbon source is still a challenge. Among biobased feedstocks, lignocellulose is a cheap, abundant and potentially sustainable carbon source. However, because of its recalcitrance, separation and depolymerization processes that have not reached industrial maturity are usually required. Integrated biorefineries utilize a holistic approach to conversion processes to minimize feedstock price and maximize resource use. This review examines the technical feasibility of merging PHA production and lignocellulose biorefining in integrated processing facilities. Among lignocellulosic feedstocks, wood is a promising carbon source due to its mature industrial infrastructure. Among the lignocellulose components, the hemicellulose fraction is the most promising feedstock for PHA production since it is underutilized and can be combined with bioethanol production from the cellulose fraction. Fractionation processes allow separate recovery of cellulose, hemicellulose and lignin, to which PHA can be added as a co-product.

Model Study To Assess Softwood Hemicellulose Hydrolysates as the Carbon Source for PHB Production in Paraburkholderia sacchari IPT 101.

Softwood hemicellulose hydrolysates are a cheap source of sugars that can be used as a feedstock to produce polyhydroxybutyrates (PHB), which are biobased and compostable bacterial polyesters. To assess the potential of the hemicellulosic sugars as a carbon source for PHB production, synthetic media containing softwood hemicellulose sugars (glucose, mannose, galactose, xylose, arabinose) and the potentially inhibitory lignocellulose degradation products (acetic acid, 5-hydroxymethylfurfural (HMF), furfural, and vanillin) were fermented with the model strain Paraburkholderia sacchari IPT 101. Relative to pure glucose, individual fermentation for 24 h with 20 g/L mannose or galactose exhibited maximum specific growth rates of 97% and 60%, respectively. On the other hand, with sugar mixtures of glucose, mannose, galactose, xylose, and arabinose, the strain converted all sugars simultaneously to reach a maximum PHB concentration of 5.72 g/L and 80.5% PHB after 51 h. The addition of the inhibitor mixture at the following concentration, sodium acetate (2.11 g/L), HMF (0.67 g/L), furfural (0.66 g/L), and vanillin (0.93 g/L), to the sugar mixture stopped the growth entirely within 24 h. Individually, the inhibitors either had no effect or only reduced growth. Moreover, it was found that a bacterial inoculum with high initial cell density (optical density, OD ≥ 5.6) could overcome the growth inhibition to yield an OD of 13 within 24 h. Therefore, softwood hemicellulose sugars are viable carbon sources for PHB production. Nevertheless, real softwood hemicellulose hydrolysates need detoxification or a high inoculum to overcome inhibitory effects and allow bacterial growth.

Fibre size does not appear to influence the ease of enzymatic hydrolysis of organosolv-pretreated softwoods.

To determine the effect of fibre size on enzymatic hydrolysis, organosolv-pretreated lodgepole pine was size-fractionated into six substrates ranging in average size from 0.20 to 3.4mm. Other than the fines fraction (<0.2mm) which contained most of the lignin, the fractionated substrates were more readily hydrolyzed than the original substrate with nearly complete hydrolysis after 72 h at 5 FPU g(-1) cellulose. Surprisingly, fibre size was found to have little influence on enzymatic hydrolysis likely due to similarities in the substrates' chemical composition, accessible surface area, cellulose crystallinity and degree of polymerization. To determine the influence of the fines on enzymatic hydrolysis, their content was artificially increased (from 8.9% to 55.4%) however; this did not have a noticeable effect. These results show that within the range of fibre sizes tested, other substrate characteristics likely play a more significant role in the ease of hydrolysis of pretreated substrates.

The effects of increasing swelling and anionic charges on the enzymatic hydrolysis of organosolv-pretreated softwoods at low enzyme loadings.

Organosolv-pretreated Lodgepole pine substrates were physically and chemically treated to increase their hydrophilicity and swelling as these are two substrate attributes which have been shown to improve cellulolytic hydrolysis. Surprisingly, mechanical treatment of the organosolv-treated substrates by PFI-mill refining did not significantly increase hydrolysis yields despite decreases in particle size and crystallinity and increases in swelling. However, sulfonation of the substrate did, significantly, increase enzymatic hydrolysis at loadings of both 5 and 2.5 FPU g(-1) cellulose (from 80% to 95% and from 35% to 80%, respectively). In addition, sulfonation resulted in an increase in the amount of free enzymes detected during the course of hydrolysis to a maximum of 80% after 72 h. This suggested that the beneficial effects of sulfonation were primarily due to a decrease in the non-specific binding of the cellulases to the lignin.

The effect of varying organosolv pretreatment chemicals on the physicochemical properties and cellulolytic hydrolysis of mountain pine beetle-killed lodgepole pine.

Mountain pine beetle-killed lodgepole pine (Pinus contorta) chips were pretreated using the organosolv process, and their ease of subsequent enzymatic hydrolysis was assessed. The effect of varying pretreatment chemicals and solvents on the substrate's physicochemical characteristics was also investigated. The chemicals employed were MgCl2, H2SO4, SO2, and NaOH, and the solvents were ethanol and butanol. It was apparent that the different pretreatments resulted in variations in both the chemical composition of the solid and liquid fractions as well in the extent of cellulolytic hydrolysis (ranging from 21% to 82% hydrolysis after 12 h). Pretreatment under acidic conditions resulted in substrates that were readily hydrolyzed despite the apparent contradiction that pretreatment under alkaline conditions resulted in increased delignification (approximately 7% and 10% residual lignin for alkaline conditions versus 17% to 19% for acidic conditions). Acidic pretreatments also resulted in lower cellulose degree of polymerization, shorter fiber lengths, and increased substrate porosity. The substrates generated when butanol/water mixtures were used as the pretreatment solvent were also hydrolyzed more readily than those generated with ethanol/water. This was likely due to the limited miscibility of the solvents resulting in an increased concentration of pretreatment chemicals in the aqueous layer and thus a higher pretreatment severity.

Microbial communities in wetlands of the Athabasca oil sands: genetic and metabolic characterization.

Naphthenic acids are a complex family of naturally occurring cyclic and acyclic carboxylic acids that are present in the acidic fraction of petroleum. Naphthenic acids are acutely toxic to aquatic organisms. Previous studies showed that wetland sediments exposed to oil sands process water containing naphthenic acids had higher rates of naphthenic acid degradation in vitro compared with unexposed wetlands. In this study we compare the microbial community structures in sediments from wetlands exposed to different amounts of oil sands process water using BIOLOG, phospholipid fatty acid analysis and denaturing gradient gel electrophoresis of total bacterial DNA. Community profiles were compared using cluster analysis. BIOLOG profiles were primarily influenced by seasonal trends rather than naphthenic acids content. In contrast, phospholipid fatty acid analysis comparisons clustered communities that had higher levels of residual oil, although this association was not strong. In contrast, cluster diagrams produced from the denaturing gradient gel electrophoresis data clearly separated bacterial communities according to naphthenic acids concentrations, indicating that naphthenic acids content was a major influence on the composition of the bacterial community. In addition, denaturing gradient gel electrophoresis profiles indicated that naphthenic acids-exposed bacterial communities were homogeneous on a scale of meters, whereas unexposed (off-site) wetlands were less homogeneous.