Khabade Sarina P, Sirigiri Divijendra Natha Reddy, Ram Anshu Beulah
Department of PG Studies in Biotechnology, Nrupathunga University, Bangalore 560001, India.
Department of Biotechnology, BMS College of Engineering, Bangalore, Affiliated to Visvesvaraya Technological University, Belagavi, Karnataka 560019, India.
ACS Omega. 2024 Jan 5;9(3):3616-3624. doi: 10.1021/acsomega.3c07633. eCollection 2024 Jan 23.
l-Asparaginase (E.C. 3.5.1.1) is an indispensable analeptic anticancer enzyme used as an amalgam with additional cancer medicines for the cure of acute lymphoblastic leukemia (ALL). The presence of lAparaginase in tomato was confirmed byWestern blotting and DNA sequencing. The l-Asparaginase gene from tomato has been deposited in the NCBI database with accession number: OR736141. Crude enzyme was extracted from the fruit pulp of and the activity was determined by the Nesslerization method. Further, the crude extract was subjected to purification, and kinetic parameters were studied. The percentage yield was calculated to be 6.457, and the purification fold was 0.086. The enzyme showed maximum activity at optimum pH 7.0, optimum temperature 37 °C, and incubation time of 05 min. The Michaelis constant "" and maximum velocity "" values were determined by the Lineweaver-Burk plot, which showed a low value of 0.66 and of 3.846 IU. Cytotoxic studies were carried out for crude and purified l-asparaginase. Purified l-Asparaginase has exhibited anticancer activity against the ALL model system, K-562 cell line, comparable to that of the anticancer compound vinblastine. Hence, l-Asparaginase from the fruit extract of tomato could be used as a nutraceutical to support cancer treatment in acute lymphoblastic leukemia.
L-天冬酰胺酶(E.C. 3.5.1.1)是一种不可或缺的解毒抗癌酶,与其他抗癌药物联合用于治疗急性淋巴细胞白血病(ALL)。通过蛋白质免疫印迹法和DNA测序证实了番茄中存在L-天冬酰胺酶。来自番茄的L-天冬酰胺酶基因已保存在NCBI数据库中,登录号为:OR736141。从[具体品种]的果肉中提取粗酶,并采用奈斯勒试剂法测定其活性。此外,对粗提物进行了纯化,并研究了动力学参数。计算得到的百分产率为6.457,纯化倍数为0.086。该酶在最适pH 7.0、最适温度37℃和孵育时间5分钟时表现出最大活性。通过Lineweaver-Burk图测定了米氏常数“Km”和最大速度“Vmax”值,结果显示较低的Km值为0.66,Vmax为3.846 IU。对粗制和纯化的L-天冬酰胺酶进行了细胞毒性研究。纯化的L-天冬酰胺酶对ALL模型系统K-562细胞系表现出抗癌活性,与抗癌化合物长春碱相当。因此,番茄果实提取物中的L-天冬酰胺酶可作为一种营养保健品,用于支持急性淋巴细胞白血病的癌症治疗。