A green technology of biodiesel production focuses on the use of enzymes as the catalyst. In enzymatic biodiesel synthesis, suitable solvent system is very essential to reduce the inhibition effects of the solvent to the enzymes. This study produced ethanol-based biodiesel from a low-cost sludge palm oil (SPO) using locally-produced Candida cylindracea lipase from fermentation of palm oil mill effluent (POME) based medium. The optimum levels of ethanol-to-SPO molar ratio and enzyme loading were found to be 4:1 and 10 U/25 g of SPO respectively with 54.4% w/w SPO yield of biodiesel and 21.7% conversion of free fatty acid (FFA) into biodiesel. Addition of tert-butanol at 2:1 tert-butanol-to-SPO molar ratio into the ethanol-solvent system increased the yield of biodiesel to 71.6% w/w SPO and conversion of FFA into biodiesel to 28.8%. The SPO and ethanol have promising potential for the production of renewable biodiesel using enzymatic-catalyzed esterification and transesterification.
Deep Eutectic Solvents (DESs) have recently emerged as a new generation of ionic liquids for lignocellulose pretreatment. However, DESs contain salt components which tend to inactivate cellulase in the subsequent saccharification process. To alleviate this problem, it is necessary to evaluate the applicability of the DESs-Cellulase system. This was accomplished in the present study by first studying the stability of cellulase in the presence of selected DESs followed by applicability evaluation based on glucose production, energy consumption and kinetic performance. Results showed that the cellulase was able to retain more than 90% of its original activity in the presence of 10% (v/v) for glycerol based DES (GLY) and ethylene glycol based DES (EG). Furthermore, both DESs system exhibited higher glucose percentage enhancement and lower energy consumption as compared to diluted alkali system. Among the two DESs studied, EG showed comparatively better kinetic performance.
Recombinant elastase strain K overexpressed from E. coli KRX/pCon2(3) was purified to homogeneity by a combination of hydrophobic interaction chromatography and ion exchange chromatography, with a final yield of 48% and a 25-fold increase in specific activity. The purified protein had exhibited a first ever reported homodimer size of 65 kDa by SDS-PAGE and MALDI-TOF, a size which is totally distinct from that of typically reported 33 kDa monomer from P. aeruginosa. The organic solvent stability experiment had demonstrated a stability pattern which completely opposed the rules laid out in previous reports in which activity stability and enhancement were observed in hydrophilic organic solvents such as DMSO, methanol, ethanol and 1-propanol. The high stability and enhancement of the enzyme in hydrophilic solvents were explained from the view of alteration in secondary structures. Elastinolytic activation and stability were observed in 25 and 50% of methanol, respectively, despite slight reduction in α-helical structure caused upon the addition of the solvent. Further characterization experiments had postulated great stability and enhancement of elastase strain K in broad range of temperatures, pHs, metal ions, surfactants, denaturing agents and substrate specificity, indicating its potential application in detergent formulation.