Lignocellulosic saccharification methods typically involve enzymatic hydrolysis. In the conventional process, 10 g of pretreated lignocellulosic material is mixed with deionized water at a specific solid-to-water ratio and adjusted to pH 6. A cellulase solution is added at 1% (w/w) concentration, and the mixture is incubated for a certain period. After hydrolysis, the reaction is stopped, and the mixture is cooled to room temperature. Centrifugation separates the enzymatic residue from the sugar-rich liquid, which is then analyzed for reducing sugars using the DNS colorimetric method.
In the ultrasonic-assisted enzymatic hydrolysis, similar conditions are maintained—temperature and pH remain constant—but an ultrasonic frequency of 20 kHz is applied. The ultrasonic treatment follows a cycle of 1 minute of sonication followed by 2 minutes of rest. After the same duration of hydrolysis, the reaction is terminated, cooled, and centrifuged as before. The resulting sugar solution is also measured for reducing sugars using the DNS method.
Ultrasonic assistance significantly impacts alkaline pretreatment. It reduces the required alkali concentration and reaction time while maintaining or even improving the decomposition rate of the raw material. Comparative experiments showed that under the same decomposition efficiency, both alkali concentration and hydrolysis time were reduced. For instance, when using ultrasonic-assisted alkaline hydrolysis, the alkali concentration dropped from 8% to 5%, and the reaction time decreased from 1.5 hours to 1 hour. This makes ultrasonic-assisted pretreatment more efficient and environmentally friendly.
The effects of ultrasonic-assisted alkaline pretreatment on the raw material components were compared with those of single alkaline hydrolysis. The results showed that the removal of lignin increased by 23.3%, and the degradation of cellulose improved by 12.1%. Alkaline pretreatment is effective in removing lignin, and the micro-vibration caused by ultrasound enhances this process further. Additionally, it increases the degradation of hemicellulose, improving the accessibility of cellulose fibers and thus boosting the enzymatic saccharification rate.
Scanning electron microscopy (SEM) was used to observe the morphological changes in corn stalks after different treatments. Untreated samples (a) showed dense and hard structures, while samples treated with only ultrasound (b) showed minimal surface changes. Alkali hydrolysis (c) caused significant structural damage, making the fibers more porous and increasing their surface area. The ultrasonic-assisted alkaline pretreatment combined these benefits, leading to a more open structure that enhances reagent accessibility and improves the overall enzymatic saccharification efficiency.
In conclusion, the use of ultrasonic-assisted alkaline pretreatment not only reduces chemical and energy consumption but also enhances the efficiency of subsequent enzymatic hydrolysis. This method shows great potential for improving the saccharification process in biofuel production.
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