A novel method for real-time estimation of insoluble solids and glucose concentrations during enzymatic hydrolysis of biomass.

The study describes a novel method using instantaneous mixing torque and rotational speed to estimate insoluble solids and glucose concentrations during enzymatic hydrolysis of biomass. This method is cost-effective for real-time monitoring and control of enzymatic hydrolysis and potentially scalable. The model was developed using biomass slurries at three solids loading (20, 30 and 45%) at various rotational speeds from 50 to 400 rpm. The results showed a significant drop in mixing torque at 12 h with high solids loading. Maximum glucose concentration (205 g/l) during hydrolysis was achieved at 45% solids loading. Insoluble solids and glucose concentration as a function of torque and rotational speeds were modeled using a modified Herschell-Bulkley model. The model describes the experimental observations with high fidelity (R2 = 0.84) and can be used for real time monitoring of many multiphase reaction systems as enzymatic hydrolysis of lignocellulosic biomass and dry grind corn ethanol processes.

Production of thermostable ß-glucosidase and CMCase by Penicillium sp: LMI01 isolated from the Amazon region

Background: Cellulolytic enzymes of microbial origin have great industrial importance because of their wide application in various industrial sectors. Fungi are considered the most efficient producers of these enzymes. Bioprospecting survey to identify fungal sources of biomass-hydrolyzing enzymes from a high-diversity environment is an important approach to discover interesting strains for bioprocess uses. In this study, we evaluated the production of endoglucanase (CMCase) and ß-glucosidase, enzymes from the lignocellulolytic complex, produced by a native fungus. Penicillium sp. LMI01 was isolated from decaying plant material in the Amazon region, and its performance was compared with that of the standard isolate Trichoderma reesei QM9414 under submerged fermentation conditions. Results: The effectiveness of LMI01 was similar to that of QM9414 in volumetric enzyme activity (U/mL); however, the specific enzyme activity (U/mg) of the former was higher, corresponding to 24.170 U/mg of CMCase and 1.345 U/mg of ß-glucosidase. The enzymes produced by LMI01 had the following physicochemical properties: CMCase activity was optimal at pH 4.2 and the ß-glucosidase activity was optimal at pH 6.0. Both CMCase and ß-glucosidase had an optimum temperature at 60°C and were thermostable between 50 and 60°C. The electrophoretic profile of the proteins secreted by LMI01 indicated that this isolate produced at least two enzymes with CMCase activity, with approximate molecular masses of 50 and 35 kDa, and ß-glucosidases with molecular masses between 70 and 100 kDa. Conclusions: The effectiveness and characteristics of these enzymes indicate that LMI01 can be an alternative for the hydrolysis of lignocellulosic materials and should be tested in commercial formulations.