Production, purification and characterization of an ionic liquid tolerant cellulase from Bacillus sp. Isolated from rice paddy field soil

Abstract

Background: Lignocellulosic biomass is a renewable, abundant, and inexpensive resource for biorefining process to produce biofuel and valuable chemicals. To make the process become feasible, it requires the use of both efficient pretreatment and hydrolysis enzymes to generate fermentable sugars. Ionic liquid (IL) pretreatment has been demonstrated to be a promising method to enhance the saccharification of biomass by cellulase enzyme; however, the remaining IL in the hydrolysis buffer strongly inhibits the function of cellulase. This study aimed to isolate a potential IL-tolerant cellulase producing bacterium to be applied in biorefining process. Result: One Bacillus sp., MSL2 strain, obtained from rice paddy field soil was isolated based on screening of cellulase assay. Its cellulase enzyme was purified and fractionated using a size exclusion chromatography. The molecular weight of purified cellulose was 48 kDa as revealed by SDS-PAGE and zymogram analysis. In the presence of the IL, 1-ethyl-3-methylimidazolium acetate ([C2mim][OAc]) concentration of 1 M, the cellulase activity retained 77.7% of non-IL condition. In addition, the optimum temperature and pH of the enzyme is 50°C and pH 6.0, respectively. However, this cellulase retained its activity more than 90% at 55°C, and pH 4.0. Kinetic analysis of purified enzyme showed that the Km and Vmax were 0.8 mg/mL and 1000 μM/min, respectively. Conclusion: The characterization of cellulase produced from MSL2 strain was described here. These properties of cellulase made this bacterial strain become potential to be used in the biorefining process. © 2015 Pontificia Universidad Católica de Valparaíso.