Computational Molecular Biology 2025, Vol.15, No.1, 53-64 http://bioscipublisher.com/index.php/cmb 60 Nowadays, when extracting polysaccharides fromCordyceps, environmental protection indicators are becoming increasingly important. The traditional methods consume both electricity and water. However, the ultrasonic-enzymatic synergistic extraction does have advantages in this regard-energy consumption can be reduced by about 40% (Yan and Wang, 2006), and the amount of solvent used is also halved. To be honest, the price of enzyme preparations is still a bit high. Fortunately, they can be recycled 3 to 4 times now, which greatly reduces the cost pressure. Laboratory data show that this green process not only saves resources, but also yields higher polysaccharide activity in the end. Of course, when industrial-scale amplification is carried out, the issue of equipment compatibility also needs to be addressed. After all, ultrasonic probes tend to wear out over time. Recently, some manufacturers have launched a new type of variable-frequency ultrasonic equipment. It is said to be both energy-saving and durable, but it is unknown how the actual production effect will be. Overall, this approach strikes a balance between efficiency and environmental protection. Although the initial investment is a bit larger, it is very cost-effective in the long run. 6 Performance Evaluation of Enzymatic and Ultrasound-Assisted Extraction 6.1 Extraction efficiency and polysaccharide purity Recently, it was discovered that the combination of ultrasound and enzymatic methods for extracting polysaccharides fromCordyceps sinensis surprisingly works well. Laboratory data show that when dealing with mycelial medium, this combination can achieve a yield of 15.43%, which is approximately 2 percentage points higher than using the enzymatic method alone. However, microwave extraction is said to have performed more vigorously (Zhu et al., 2016), but unfortunately the specific data have been concealed and not fully disclosed. Interestingly, even with the assistance of ultrasound, the effect of processing mycelium in liquid culture is significantly worse, which may be related to the difference in cell wall structure. These new methods are indeed much better than the old-fashioned hot water extraction, but the equipment investment cost will also increase accordingly. Small-scale production really needs to think twice. Purifying Cordyceps polysaccharides is by no means an easy task. Take Sephadex G-100 column chromatography as an example. Although polysaccharides can be separated by molecular weight, interestingly, polysaccharides composed of the same monosaccharides still have differences in properties. The polysaccharides extracted by ultrasound are quite special. The molecular arrangement is tighter than that obtained by other methods, which might be due to the fact that ultrasound has shaken off some impurity proteins. However, during the purification process, it was found that some small-molecule impurities were particularly stubborn and could not be dealt with merely by gel chromatography. In the laboratory, it was tried that by combining ultrasonic extraction with ion-exchange chromatography, the purity could be increased from 75% to around 90%. However, with more steps, the cost also rose accordingly. 6.2 Bioactivity evaluation The antioxidant capacity of Cordyceps polysaccharides is indeed quite remarkable, especially CMMP-1 obtained by ultrasound-assisted enzymatic method. The experimental data are quite impressive. Ultrasonic extraction alone is not bad either, and it is very effective against ABTS and hydroxyl radicals (Xiao et al., 2012). However, when it comes to immune regulation, although no direct macrophage experimental data have been observed, there is an interesting phenomenon-polysaccharides extracted by microwave have the best anti-tumor effect, which might also be related to immune regulation. It was also found in the laboratory that although polysaccharides obtained by different extraction methods all have antioxidant properties, the efficiency of eliminating different free radicals varies greatly. This might be related to their molecular configurations. Unfortunately, nowadays, fewCordyceps products on the market indicate the specific extraction process, and consumers have no idea at all about the activity of the polysaccharides they purchase. 6.3 Economic and environmental benefit analysis Nowadays, the extraction of Cordyceps polysaccharides has indeed brought many benefits through new technologies. Enzymatic and ultrasound-assisted methods not only save electricity and water, but more importantly, the polysaccharides extracted have particularly high activity, which is just right for making health
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