Posoongnoen Saijai, Thummavongsa Theera, Junthip Jatupol, Jandaruang Jinda, Preecharram Sutthidech
Division of Chemistry, Faculty of Science and Technology, Nakhon Ratchasima Rajabhat University, Nakhon Ratchasima, 30000, Thailand.
Division of Biology, Faculty of Science and Technology, Nakhon Ratchasima Rajabhat University, Nakhon Ratchasima, 30000, Thailand.
Sci Rep. 2025 Jul 15;15(1):25499. doi: 10.1038/s41598-025-11653-z.
The α-amylase from Cucurbita moschata Duchesne seeds (CmAmy) was purified using ammonium sulfate fractionation, followed by β-cyclodextrin sepharose 6B affinity chromatography. The enzyme exhibited a specific activity of 475.00 U/mg, resulting in a 208.54-fold and a 30.89% yield. The optimal pH and temperature for CmAmy were determined to be 7.0 and 60 °C, respectively. The enzyme demonstrated high stability across a temperature range of 40-80 °C and a pH range of 5-9. The CmAmy has been identified as an eco-friendly candidate for bioethanol production by utilizing cassava and rice waste. The optimal condition for sugar production through hydrolysis with CmAmy involved an enzyme concentration of 7.5 mU, a substrate concentration of 2.0% w/v, and a hydrolysis duration of 4 h for cassava waste. For rice waste, the optimal parameters consisted of an enzyme amount of 10 mU, a substrate concentration of 1.5% w/v, and a hydrolysis time of 3 h. The resulting sugars were fermented using Saccharomyces cerevisiae to yield bioethanol, achieving maximum concentrations of 19.6 g/L (0.98 g ethanol/g cassava waste) and 20.1 g/L (1.33 g ethanol/g rice waste) for cassava and rice waste, respectively. This investigation highlights the potential of CmAmy for eco-friendly bioethanol production, offering advantages of simplicity, cost-effectiveness, and sustainability.
采用硫酸铵分级沉淀法，随后进行β-环糊精琼脂糖6B亲和层析，对南瓜种子中的α-淀粉酶（CmAmy）进行了纯化。该酶的比活性为475.00 U/mg，纯化倍数为208.54倍，产率为30.89%。CmAmy的最适pH值和温度分别确定为7.0和60℃。该酶在40-80℃的温度范围和5-9的pH范围内表现出高稳定性。CmAmy已被确定为利用木薯和水稻废料生产生物乙醇的环保候选酶。用CmAmy水解生产糖的最佳条件是：木薯废料的酶浓度为7.5 mU，底物浓度为2.0% w/v，水解时间为4小时。对于水稻废料，最佳参数为酶量10 mU，底物浓度1.5% w/v，水解时间3小时。所得的糖用酿酒酵母发酵生产生物乙醇，木薯废料和水稻废料分别达到最大浓度19.6 g/L（0.98 g乙醇/g木薯废料）和20.1 g/L（1.33 g乙醇/g水稻废料）。本研究突出了CmAmy在环保生物乙醇生产方面的潜力，具有操作简单、成本效益高和可持续性等优点。
