Fungal delignification can be a feasible process to pretreat biomass for bioethanol production if its performance is improved in terms of efficiency through a few modifications. The aim of this study was to enhance the biodelignification pretreatment of rice straw using laccase in the presence of ionic liquid (1-Allyl-3-methylimidazolium chloride, [AMIM]Cl) or surfactant (TritonX-100). Addition of 750 mg/L [AMIM]Cl and 500 mg/L TritonX-100 increases the lignin removal to 18.49% and 31.79%, which is higher than that of laccase only (11.97%). The enzymatic saccharification process was carried out based on different strategies. The highest cellulose conversion, 40.96%, 38.24%, and 37.91%, was obtained after 72 h of enzymatic saccharification when the substrate was washed with distilled water after pretreatment of rice straw with laccase + TritonX-100, laccase + [AMIM]Cl, and laccase only, respectively. In addition, the morphology and structure changes of pretreated and untreated rice straw were studied. Both surface area and cellulose crystallinity are substantially altered after laccase + [AMIM]Cl and laccase + TritonX-100 pretreatment. Enhanced saccharification efficiency of rice straw was achieved by laccase pretreatment with ionic liquid or surfactant in a single system.

The effects of chymosin on the physicochemical and hydrolysis characteristics of casein micelles and individual caseins were investigated. Adding 0.03 units of chymosin/mL led to the casein micelles in skim milk coagulating after a 3 h incubation period at 30 °C. SDS–PAGE investigation showed that β-CN, κ-CN, α_s-CN, and a portion of β-lactoglobulin (β-LG) in the milk supernatant fraction (MSF) were precipitated into the milk pellet fraction (MPF). The mean particle size of the MSF with chymosin decreased from 254.4 nm to 179.2 nm after a 3 h incubation period. Mass spectrometry and SDS–PAGE analysis suggested that chymosin hydrolyzed individual β-CN, κ-CN, and α_s-CN, but not β-LG. Chymosin hydrolysis led to a decrease in the molecular weights of the hydrolyzed β-CN, κ-CN, and α_s-CN. Particle size analysis indicated that there was no difference in the particle size distribution of hydrolyzed β-CN and α_s-CN. Moreover, our outcomes demonstrated that the hydrolysis of κ-CN by chymosin occurs before that of β-CN and α_s-CN.