Computational Molecular Biology 2025, Vol.15, No.1, 53-64 http://bioscipublisher.com/index.php/cmb 59 sufficient. Interestingly, adding a 5-minute ultrasound in the middle of enzymatic hydrolysis can increase the yield by another 5%, but the operation is too troublesome and industrial production may not be able to handle it. Figure 2 SEM micrographs of the surface structure of U. lactuca samples (1000×) (Adopted from Wang et al., 2024a) Image caption: (A) untreated, (B) ultrasound-assisted hot water treatment and (C) ultrasound-assisted cellulase treatment (Adopted from Wang et al., 2024a) When conducting ultrasound-enzymatic co-extraction, temperature and pH are two parameters that people both love and hate. Enzymes are particularly delicate-they slack off when the temperature drops to 40℃and stop working directly when it exceeds 60℃. The pH value is even more picky. A difference of 0.5 May cause the activity to drop by half (Yang et al., 2017). But interestingly, ultrasound works best at around 55℃, which is also the "comfort zone" of most enzymes. Repeated debugging in the laboratory found that when the pH was controlled at 5.5 and the temperature was maintained in the sweet spot range of 50-55℃, the yield of polysaccharides could be increased by 25% compared with the conventional conditions. However, in actual operation, the temperature of the reaction system will continuously rise with the ultrasonic waves, so a sensitive temperature control system needs to be equipped for real-time regulation. Recently, it was also discovered that the optimal pH for Cordyceps sinensis from different origins actually varies slightly. This finding requires us to re-optimize our extraction process. 5.3 Advantages of synergistic extraction The combination of ultrasound and enzymatic methods for extracting polysaccharides is indeed much more effective than doing it alone. Let's start with the output. The combined use can extract 20% to 30% more polysaccharides. This has been verified several times in the laboratory. What is even more remarkable is that the polysaccharides proposed in this way are particularly "clean" with far fewer impurities. However, the most surprising thing is that the structure of the polysaccharides was hardly damaged-those key active groups were all well preserved. Of course, in actual operation, it should be noted that the ultrasonic power should not be too high; otherwise, it may instead shatter the polysaccharide molecules. Recently, it has also been found that ultrasonic treatment followed by enzyme addition yields better results than the reverse operation. This might be because the enzymes are more likely to function when the cell walls are loosened by vibration. Although the equipment investment is a bit high, considering that the purification process can be simplified, it is still quite cost-effective in the long run.
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