Podzimek Tomáš, Loužecká Karolína, Urbánková Veronika, Beránek Jan, Lipovová Petra, Benešová Eva
Department of Biochemistry and Microbiology, University of Chemistry and Technology, Technická 5, 166 28, Prague 6, Czech Republic.
Folia Microbiol (Praha). 2025 Jul 11. doi: 10.1007/s12223-025-01296-y.
L-Asparaginases are enzymes known for decades due to their use in medicine for the treatment of acute lymphoblastic leukemia. Recently, they have also found application in the food industry, and other possibilities are emerging in the treatment of infectious diseases or in the design of biosensors. For this reason, an ongoing effort has been made to find and characterize new enzymes with properties suitable for these specific applications. In this work, L-asparaginase from Paenibacillus thiaminolyticus (isoenzyme 1) belonging to the least explored group of L-asparaginases derived from L-asparaginase from Rhizobium etli was recombinantly produced with high yields (335 mg per L of culture medium) in E. coli cells and characterized: K = (26 ± 8) mmol L, pH = 10.3, without L-glutaminase and urease activity. A probable oligomeric structure (homodimer) was derived by computer modeling and confirmed by gel chromatography experiments. The results of this work extend the current limited knowledge about the poorly described class of R. etli L-asparaginases. Moreover, this L-asparaginase exhibits suitable properties for use in biosensor construction because of the high yields during recombinant production, K value, stability, and absence of L-glutaminase activity.
天冬酰胺酶作为一种酶已为人所知数十年，因其在医学上用于治疗急性淋巴细胞白血病。最近，它们在食品工业中也有应用，并且在传染病治疗或生物传感器设计方面也出现了其他可能性。因此，人们一直在努力寻找和鉴定具有适合这些特定应用特性的新酶。在这项工作中，来自解硫胺芽孢杆菌（同工酶1）的天冬酰胺酶属于源自根瘤菌天冬酰胺酶的研究最少的天冬酰胺酶组，在大肠杆菌细胞中以高产率（每升培养基335毫克）重组产生并进行了表征：K =（26±8）毫摩尔/升，pH = 10.3，无L-谷氨酰胺酶和脲酶活性。通过计算机建模推导并经凝胶色谱实验证实了可能的寡聚结构（同型二聚体）。这项工作的结果扩展了目前关于描述较少的根瘤菌天冬酰胺酶类别的有限知识。此外，由于重组生产过程中的高产率、K值、稳定性以及缺乏L-谷氨酰胺酶活性，这种天冬酰胺酶具有适合用于构建生物传感器的特性。
