这项工作的目的是使用台式生物反应器通过不同的补料分批培养策略优化大肠杆菌中欧文氏菌胡萝卜素-天冬酰胺酶 II 酶的生产，并评估重组酶对不同急性淋巴细胞白血病细胞系的治疗潜力。使用 DO-stat 补料策略并在培养 18 小时内进行诱导的培养物中获得了最高的酶活性（∼98,000 U/L）。在这些实验条件下，每个底物的重组L-天冬酰胺酶II (rASNase)产量、每个生物量的rASNase产量和生产率的最大值约为1204 U/g葡萄糖、3660 U/g细胞和3260 U/(L·h)，分别。该条件对于实现重组酶的高产率是有效的，该重组酶被纯化并用于体外抗白血病潜力测试。在所有测试的白血病细胞系中，RS4;11 对 rASNase 的敏感性最高，IC50 值约为 0.0006 U/mL，细胞凋亡率超过 70%。研究表明，所使用的培养策略可有效获得 rASNase 的高产量和生产力，该蛋白具有治疗潜力，因为该蛋白具有细胞毒活性和诱导细胞凋亡的作用。© 2024 作者。由美国化学会出版。

The aims of this work were to optimize the production of Erwinia carotovoral-asparaginase II enzyme in Escherichia coli by different fed-batch cultivation strategies using a benchtop bioreactor and to evaluate the therapeutic potential of the recombinant enzyme against different acute lymphoblastic leukemia cell lines. The highest enzyme activities (∼98,000 U/L) were obtained in cultures using the DO-stat feeding strategy with induction in 18 h of culture. Under these experimental conditions, the maximum values for recombinant l-asparaginase II (rASNase) yield per substrate, rASNase yield per biomass, and productivity were approximately 1204 U/gglucose, 3660 U/gcells, and 3260 U/(L·h), respectively. This condition was efficient for achieving high yields of the recombinant enzyme, which was purified and used in in vitro antileukemic potential tests. Of all the leukemic cell lines tested, RS4;11 showed the highest sensitivity to rASNase, with an IC50 value of approximately 0.0006 U/mL and more than 70% apoptotic cells. The study demonstrated that the cultivation strategies used were efficient for obtaining high yield and productivity of rASNase with therapeutic potential inasmuch as cytotoxic activity and induction of apoptosis were demonstrated for this protein.© 2024 The Authors. Published by American Chemical Society.