Environmentally Benign Fast-Degrading Conductive Composites.

An environmentally benign conductive composite that rapidly degrades in the presence of warm water via enzyme-mediated hydrolysis is described. This represents the first time that hydrolytic enzymes have been immobilized onto eco-friendly conductive carbon sources with the express purpose of degrading the encapsulating biodegradable plastic. Amano Lipase (AL)-functionalized carbon nanofibers (CNF) were compounded with polycaprolactone (PCL) to produce the composite film CNFAL-PCL (thickness ∼ 600 µm; CNFAL = 20.0 wt %). To serve as controls, films of the same thickness were also produced, including CNF-AL5-PCL (CNF mixed with AL and PCL; CNF = 19.2 wt % and AL = 5.00 wt %), CNF-PCL (CNF = 19.2 wt %), ALx-PCL (AL = x = 1.00 or 5.00 wt %), and PCL. The electrical performance of the CNF-containing composites was measured, and conductivities of 14.0 ± 2, 22.0 ± 5, and 31.0 ± 6 S/m were observed for CNFAL-PCL, CNF-AL5-PCL, and CNF-PCL, respectively. CNFAL-PCL and control films were degraded in phosphate buffer (2.00 mg/mL film/buffer) at 50 °C, and their average percent weight loss (Wtavg%) was recorded over time. After 3 h CNFAL-PCL degraded to a Wtavg% of 90.0% and had completely degraded after 8 h. This was considerably faster than CNF-AL5-PCL, which achieved a total Wtavg% of 34.0% after 16 days, and CNF-PCL, which was with a Wtavg% of 7.00% after 16 days. Scanning electron microscopy experiments (SEM) found that CNFAL-PCL has more open pores on its surface and that it fractures faster during degradation experiments which exposes the interior enzyme to water. An electrode made from CNFAL-PCL was fabricated and attached to an AL5-PCL support to form a fast-degrading thermal sensor. The resistance was measured over five cycles where the temperature was varied between 15.0-50.0 °C. The sensor was then degraded fully in buffer at 50 °C over a 48 h period.

Woody biomass as a potential feedstock for fermentative gaseous biofuel production.

Biogas and biohydrogen are compatible gaseous biofuels that can be blended with natural gas for reticulated fuel supply to reduce greenhouse gas emissions. Sustainably grown woody biomass is emerging as a potential feedstock in the production of biofuels. Woody biomass is widely available, uses non-arable land for plantation, does not require synthetic fertilisers to grow and acts as a carbon sink. The cellulose and hemicellulose fractions of wood are renewable sources of sugars that can be used for fermentative production of gaseous biofuels. However, widespread use of wood as a gaseous biofuel feedstock is constrained due to the recalcitrant nature of wood to enzymatic hydrolysis. Pretreatment makes cellulose and hemicellulose accessible to microbial enzymes to produce fermentable sugars. Here we review wood composition, its structure and different pretreatment techniques in the context of their effects on deconstruction of wood to improve hydrolysis and fermentative gaseous fuel production. The anaerobic digestion of pretreated wood for biogas and dark fermentation for biohydrogen production are discussed with reference to gas yields. Key advancements in lab-scale research are described for pretreatments and for pure, co- and mixed culture fermentations. Limitations to yield improvements are identified and future perspectives and prospects of gaseous biofuel production from woody biomass are discussed, with reference to new developments in engineered biocatalysts and process integration.

Synthesis of graft copolymers of chitosan-poly(caprolactone) by lipase catalysed reactive extrusion.

Chitin is an abundant natural polymer and its deacetylated derivative chitosan has been a focus for the development of biobased, biocompatible and antimicrobial materials. In this work, a green and scalable route to grafting polycaprolactone (PCL) to chitosan using an enzyme catalysed reactive extrusion process is described. FTIR, 1H and 13C NMR spectroscopy and HSQC analysis confirm grafting of PCL to chitosan and show differences in the grafting pattern obtained using two commercially produced lipase enzymes from Candida antarctica (CALB® and NovoCor®). The thermostable NovoCor enzyme gave a much higher grafting yield (96.3%) than the less thermostable CALB enzyme (5.90%). In the esterification reaction, CALB preferentially catalyses reaction on primary OH groups at the C-6 position of chitosan, whereas NovoCor catalyses on the secondary OH groups of chitosan at the C-3 position. This is related to the differences in the selectivity of the two lipase enzymes. The control synthesized without enzyme did not show any grafting reaction. The degree of crystallinity and thermal stability of the lipase catalysed copolymer was reduced compared to unmodified chitosan. Moreover, the PCL grafted chitosan produced by a solvent free reactive extrusion route retained antimicrobial property against E.coli. Such grafted co-polymers may have applications in the controlled release coatings and tissue culture surfaces.

A mild thermomechanical process for the enzymatic conversion of radiata pine into fermentable sugars and lignin.

BACKGROUND: Conversion of softwoods into sustainable fuels and chemicals is important for parts of the world where softwoods are the dominant forest species. While they have high theoretical sugar yields, softwoods are amongst the most recalcitrant feedstocks for enzymatic processes, typically requiring both more severe pretreatment conditions and higher enzyme doses than needed for other lignocellulosic feedstocks. Although a number of processes have been proposed for converting softwoods into sugars suitable for fuel and chemical production, there is still a need for a high-yielding, industrially scalable and cost-effective conversion route. RESULTS: We summarise work leading to the development of an efficient process for the enzymatic conversion of radiata pine (Pinus radiata) into wood sugars. The process involves initial pressurised steaming of wood chips under relatively mild conditions (173 °C for 3-72 min) without added acid catalyst. The steamed chips then pass through a compression screw to squeeze out a pressate rich in solubilised hemicelluloses. The pressed chips are disc-refined and wet ball-milled to produce a substrate which is rapidly saccharified using commercially available enzyme cocktails. Adding 0.1% polyethylene glycol during saccharification was found to be particularly effective with these substrates, reducing enzyme usage to acceptable levels, e.g. 5 FPU/g OD substrate. The pressate is separately hydrolysed using acid, providing additional hemicellulose-derived sugars, for an overall sugar yield of 535 kg/ODT chips (76% of theoretical). The total pretreatment energy input is comparable to other processes, with the additional energy for attrition being balanced by a lower thermal energy requirement. This pretreatment strategy produces substrates with low levels of fermentation inhibitors, so the glucose-rich mainline and pressate syrups can be fermented to ethanol without detoxification. The lignin from the process remains comparatively unmodified, as evident from the level of retained ß-ether interunit linkages, providing an opportunity for conversion into saleable co-products. CONCLUSIONS: This process is an efficient route for the enzymatic conversion of radiata pine, and potentially other softwoods, into a sugar syrup suitable for conversion into fuels and chemicals. Furthermore, the process uses standard equipment that is largely proven at commercial scale, de-risking process scale-up.

Micromorphological changes and mechanism associated with wet ball milling of Pinus radiata substrate and consequences for saccharification at low enzyme loading.

In this work, substrates prepared from thermo-mechanical treatment of Pinus radiata chips were vibratory ball milled for different times. In subsequent enzymatic hydrolysis, percent glucan conversion passed through a maximum value at a milling time of around 120min and then declined. Scanning electron microscopy revealed breakage of fibers to porous fragments in which lamellae and fibrils were exposed during ball milling. Over-milling caused compression of the porous fragments to compact globular particles with a granular texture, decreasing accessibility to enzymes. Carbon-13 NMR spectroscopy showed partial loss of interior cellulose in crystallites, leveling off once fiber breakage was complete. A mathematical model based on observed micromorphological changes supports ball milling mechanism. At a low enzyme loading of 2FPU/g of substrate and milling time of 120min gave a total monomeric sugar yield of 306g/kg of pulp which is higher than conventional pretreatment method such as steam exploded wood.

Green route to modification of wood waste, cellulose and hemicellulose using reactive extrusion.

A large volume of wood waste is produced in timber processing industry which traditionally used in low value applications. Here, value addition to the wood waste (Sander dust) and cellulose, hemicellulose isolated thereof by functionalisation using cyclic anhydrides in a solvent-free and green reactive extrusion process is reported. The effect of extrusion temperature, catalyst and different weight ratios of Sander dust (SD):succinic anhydride (SA) on the esterification reaction is evaluated. The esterified products were characterised by the acid value, degree of substitution (DS), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), solid state (13)C NMR and thermo-gravimetric analysis (TGA). Under optimum extrusion conditions, mixed esters are formed, with highest acid value obtained for succinylation of cellulose (0.122 g/g at DS of 0.350) which is two times higher compared to succinylated SD (0.059 g/g at a weight gain of 0.452) and hemicellulose (0.043 g/g at DS of 0.290). The reactivity trend for individual anhydride was: (1) SA-Cellulose>SD>hemicellulose; (2) maleic anhydride (MA)-SD>hemicellulose>cellulose and (3) dodecenyl succinic anhydride (DDSA)-SD ≈ cellulose ≫ hemicellulose. The pendant free carboxyl groups generated through functionalisation of wood waste, cellulose and hemicellulose without the presence of polymeric carriers will allow more tailored or targeted modification of wood-plastic composites.

Improved saccharification of steam-exploded Pinus radiata on supplementing crude extract of Penicillium sp.

Commercially available enzymes do not contain all the necessary softwood-specific accessory enzymes to obtain high saccharification efficiency. In this work, six saprophytic fungi obtained from Pinus radiata plantation site were screened for the putative softwood-specific accessory enzyme, ß-mannanase. A Penicillium sp. was found to produce ß-mannanase in both solid (31.6 units/g of dry biomass) and liquid (117 units/g of dry biomass) cultures using locust bean gum as an inducer after 2 weeks of incubation. The saccharification of steam-exploded Pinus radiata was 7.8 % w/w improved when the crude extract of Penicillium sp. was added to a mixture of commercial enzymes.

A mathematical model for the inhibitory effects of lignin in enzymatic hydrolysis of lignocellulosics.

A new model for enzymatic hydrolysis of lignocellulosic biomass distinguishes causal influences from enzyme deactivation and restrictions on the accessibility of cellulose. It focuses on calculating the amount of unreacted cellulose at cessation of enzyme activity, unlike existing models that were constructed for calculating the time dependence of conversion. There are three adjustable parameters: (1) 'occluded cellulose' is defined as cellulose that cannot be hydrolysed regardless of enzyme loading or incubation time, (2) a 'characteristic enzyme loading' is sufficient to hydrolyse half of the non-occluded cellulose, (3) a 'mechanism index' measures deviations from first-order kinetics. This model was used to predict that the optimal incubation temperature is lower for lignocellulosics than for pure cellulose. For steam-exploded pine wood after 96h incubation, occluded cellulose was 24% and 26% at 30°C and 50°C, and the characteristic enzyme loadings were 10 and 18FPU/g substrate, respectively.

Optimizing the enzyme loading and incubation time in enzymatic hydrolysis of lignocellulosic substrates.

A mathematical model for costing enzymatic hydrolysis of lignocellulosics is presented. This model is based on three variable parameters describing substrate characteristics and three unit costs for substrate, enzymes and incubation. The model is used to minimize the cost of fermentable sugars, as intermediate products on the route to ethanol or other biorefinery products, by calculating optimized values of enzyme loading and incubation time. This approach allows comparisons between substrates, with processing conditions optimized independently for each substrate. Steam-exploded pine wood was hydrolyzed in order to test the theoretical relationship between sugar yield and processing conditions.