Abstract：
To harness the diverse industrial applications of cellulase, including its use in the food, pulp, textile, agriculture, and biofuel sectors, this study focused on the high-yield production of a bioactive insect-derived endoglucanase, Monochamus saltuarius glycoside hydrolase family 5 (MsGHF5). MsGHF5 was introduced into the genome of Kluyveromyces lactis to maintain expression stability, and mass production of the enzyme was induced using fed-batch fermentation. After 40 h of cultivation, recombinant MsGHF5 was successfully produced in the culture broth, with a yield of 29,000 U/L, upon galactose induction. The optimal conditions for the activity of purified MsGHF5 were determined to be a pH of 5 and a temperature of 35 °C, with the presence of ferrous ions enhancing the enzymatic activity by up to 1.5-fold. Notably, the activity of MsGHF5 produced in K. lactis was significantly higher than that produced in Escherichia coli, suggesting that glycosylation is crucial for the functional performance of the enzyme. This study highlights the potential use of K. lactis as a host for the production of bioactive MsGHF5, thus paving the way for its application in various industrial sectors.
摘要：
为了利用纤维素酶的各种工业应用，包括它在食物中的使用，纸浆,纺织品,农业,和生物燃料部门，这项研究的重点是高产生产生物活性昆虫来源的内切葡聚糖酶，单香菜GH家族5（MsGHF5）。将MsGHF5导入乳酸克鲁维酵母基因组以维持表达稳定性,并使用补料分批发酵诱导酶的大量生产。培养40小时后，重组MsGHF5在培养液中成功生产，产量为29,000U/L，在半乳糖诱导。确定纯化的MsGHF5活性的最佳条件为pH为5，温度为35°C，亚铁离子的存在使酶活性提高了1.5倍。值得注意的是,在K.lactis中产生的MsGHF5的活性显着高于在大肠杆菌中产生的MsGHF5，表明糖基化对酶的功能性能至关重要。这项研究强调了K.lactis作为生产生物活性MsGHF5的宿主的潜在用途，从而为其在各个工业部门的应用铺平了道路。
