Cellulose nanostructures obtained using enzymatic cocktails with different compositions.

Cellulose nanostructures obtained from lignocellulosic biomass by the enzymatic route can offer advantages in terms of material properties and processing sustainability. However, most of the enzymatic cocktails commonly used in the saccharification of biomass are designed to promote the complete depolymerization of the cellulose structure into soluble sugars. Here, investigation was made of the way that the action of different commercially available cellulase enzyme cocktails can affect the production of nanocellulose. For this, enzymatic cocktails designed for complete or partial saccharification were compared, using eucalyptus cellulose pulp as a model feedstock. The results showed that all the enzymatic cocktails were effective in the formation of nanocellulose structures, with the complete saccharification enzymes being more efficient in promoting the coproduction of glucose (36.5 g/L, 87% cellulose conversion). The presence of auxiliary enzymes, especially xylanases, acted cooperatively to favor the production of nanostructures with higher crystallinity (up to 79%), higher surface charge (zeta potential up to -30.9 mV), and more uniform dimensions within the size range of cellulose nanocrystals (80 to 350 nm). Interestingly, for the enzymatic cocktails designed for partial saccharification, the xylanase activity was more important than the endoglucanase activity in the production of nanocellulose with improved properties. The findings showed that the composition of the enzymatic cocktails already used for complete biomass saccharification can be suitable for obtaining nanocellulose, together with the release of a glucose stream, in a format compatible with the biorefinery concept.

Low-cost bacterial nanocellulose-based interdigitated biosensor to detect the p53 cancer biomarker.

Low-cost sensors to detect cancer biomarkers with high sensitivity and selectivity are essential for early diagnosis. Herein, an immunosensor was developed to detect the cancer biomarker p53 antigen in MCF7 lysates using electrical impedance spectroscopy. Interdigitated electrodes were screen printed on bacterial nanocellulose substrates, then coated with a matrix of layer-by-layer films of chitosan and chondroitin sulfate onto which a layer of anti-p53 antibodies was adsorbed. The immunosensing performance was optimized with a 3-bilayer matrix, with detection of p53 in MCF7 cell lysates at concentrations between 0.01 and 1000 Ucell. mL-1, and detection limit of 0.16 Ucell mL-1. The effective buildup of the immunosensor on bacterial nanocellulose was confirmed with polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) and surface energy analysis. In spite of the high sensitivity, full selectivity with distinction of the p53-containing cell lysates and possible interferents required treating the data with a supervised machine learning approach based on decision trees. This allowed the creation of a multidimensional calibration space with 11 dimensions (frequencies used to generate decision tree rules), with which the classification of the p53-containing samples can be explained.

A new approach to obtain cellulose nanocrystals and ethanol from eucalyptus cellulose pulp via the biochemical pathway.

The feasibility of integration of cellulosic ethanol production with the manufacture of cellulose nanofibers (CNF) and cellulose nanocrystals (CNC) was evaluated using eucalyptus cellulose pulp as feedstock and employing the biochemical route alone. For the enzymatic hydrolysis step, experimental central composite design (CCD) methodology was used as a tool to evaluate the effects of solids loading (SL) and enzymatic loading (EL) on glucose release and cellulose conversion. Glucose concentrations from 45 to 125 g/L were obtained after 24 h, with cellulose conversions from 35 to 96%. Validation of the statistical model was performed at SL of 20% and EL of 10 mg protein/g, which was defined by the desirability function as the optimum condition. The sugars released were used for the production of ethanol by Saccharomyces cerevisiae, resulting in 62.1 g/L ethanol after 8 h (yield of 95.5%). For all the CCD experimental conditions, the residual solids presented CNF characteristics. Moreover, the use of a new strategy with temperature reduction from 50 to 35°C after 24 h of enzymatic hydrolysis enabled CNC to be obtained after 144 h. The CNC showed a crystallinity index of 83%, length of 260 nm, diameter of 15 nm, and aspect ratio (L/D) of 15. These characteristics are suitable for many applications, such as reinforcement in polymeric materials and other lower volume higher value bio-based products. The findings indicate the viability of obtaining ethanol and CNC using the biochemical route exclusively, potentially contributing to the future implementation of forest biorefineries. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1085-1095, 2017.