Krasotkina Julya, Borisova Anna A, Gervaziev Yuri V, Sokolov Nikolay N
Laboratory of Medical Biotechnology, Institute for Biomedical Chemistry, Russian Academy of Medical Sciences, 559-B, 10 Pogodinskay St, Moscow 119121, Russia.
Biotechnol Appl Biochem. 2004 Apr;39(Pt 2):215-21. doi: 10.1042/BA20030138.
ECAR-LANS, the recombinant L-asparaginase from Erwinia carotovora, is a prospective therapeutic enzyme for leukaemia treatment. An efficient and economical scheme was developed for the purification, cloning and expression in Eschericha coli of ECAR-LANS. More than 90% purity, complemented with 72% active enzyme recovery, was achieved with a single chromatographic purification step. The activity of purified L-asparaginase was 630 i.u./mg. The ECAR-LANS K (m) value was 98x10(-6) M for the main physiological substrate L-Asn and 3400x10(-6) M for L-Gln. ECAR-LANS was found to have low relative glutaminase activity (1.2%) at physiological concentrations of L-Asn and L-Gln in blood. Kinetic studies of ECAR-LANS showed that the recombinant asparaginase combined the main advantages of Erw. chrysanthemi and E. coli L-asparaginases II, currently used in the treatment of acute lymphoblastic leukaemia.
来自胡萝卜软腐欧文氏菌的重组L-天冬酰胺酶（ECAR-LANS）是一种用于白血病治疗的潜在治疗性酶。开发了一种高效且经济的方案，用于ECAR-LANS在大肠杆菌中的纯化、克隆和表达。通过单步色谱纯化步骤，实现了超过90%的纯度，活性酶回收率为72%。纯化的L-天冬酰胺酶活性为630国际单位/毫克。ECAR-LANS对主要生理底物L-天冬酰胺的K（m）值为98×10⁻⁶ M，对L-谷氨酰胺的K（m）值为3400×10⁻⁶ M。发现在血液中L-天冬酰胺和L-谷氨酰胺的生理浓度下，ECAR-LANS具有较低的相对谷氨酰胺酶活性（1.2%）。ECAR-LANS的动力学研究表明，重组天冬酰胺酶结合了目前用于治疗急性淋巴细胞白血病的菊欧文氏菌和大肠杆菌L-天冬酰胺酶II的主要优点。
