Dhankhar Rakhi, Kawatra Anubhuti, Gupta Vatika, Mohanty Aparajita, Gulati Pooja
Medical Microbiology and Bioprocess Technology Laboratory, Department of Microbiology, Maharshi Dayanand University, Rohtak, Haryana India.
Molecular Biology and Genetic Engineering Laboratory, School of Biotechnology, Jawaharlal Nehru University, New Delhi, India.
3 Biotech. 2022 Sep;12(9):220. doi: 10.1007/s13205-022-03292-2. Epub 2022 Aug 12.
Arginine deiminase (ADI), a promising anticancer enzyme from is currently in phase III of clinical trials for the treatment of arginine auxotrophic tumors. However, it has been associated with several drawbacks in terms of low stability at human physiological conditions, high immunogenicity, hypersensitivity and systemic toxicity. In our previous work, 24 was identified as a potent producer of ADI with optimum activity under physiological conditions. In the present study, phylogenetic analysis of microbial ADIs indicated ADI (PfADI) to be closely related to experimentally characterized ADIs of sp. with proven anticancer activity. Immunoinformatics analysis was performed indicating lower immunogenicity of PfADI than MhADI ( ADI) both in terms of number of linear and conformational B-cell epitopes and T-cell epitope density. Overall antigenicity and allergenicity of PfADI was also lower as compared to MhADI, suggesting the applicability of PfADI as an alternative anticancer biotherapeutic. Hence, in vitro experiments were performed in which the ADI coding gene of was cloned and expressed in BL21. Recombinant ADI of was purified, characterized and its anticancer activity was assessed. The enzyme was stable at human physiological conditions (pH 7 and 37 °C) with m of 1.90 mM. PfADI was found to effectively inhibit the HepG2 cells with an IC50 value of 0.1950 IU/ml. Therefore, the current in silico and in vitro studies establish PfADI as a potential anticancer drug candidate with improved efficacy and low immunogenicity.
The online version contains supplementary material available at 10.1007/s13205-022-03292-2.
精氨酸脱亚氨酶（ADI）是一种来自[具体来源未提及]的有前景的抗癌酶，目前正处于治疗精氨酸营养缺陷型肿瘤的III期临床试验阶段。然而，它在人类生理条件下稳定性低、免疫原性高、过敏反应和全身毒性方面存在几个缺点。在我们之前的工作中，[具体微生物未提及]24被鉴定为在生理条件下具有最佳活性的ADI高效生产者。在本研究中，对微生物ADI进行系统发育分析表明，PfADI（[具体微生物来源未提及]的ADI）与具有已证实抗癌活性的[具体微生物属未提及]种的经实验表征的ADI密切相关。进行了免疫信息学分析，结果表明，就线性和构象性B细胞表位数量以及T细胞表位密度而言，PfADI的免疫原性低于MhADI（[具体微生物来源未提及]的ADI）。与MhADI相比，PfADI的总体抗原性和致敏性也较低，这表明PfADI作为一种替代抗癌生物治疗剂的适用性。因此，进行了体外实验，其中[具体微生物来源未提及]的ADI编码基因被克隆并在大肠杆菌BL21中表达。纯化并表征了[具体微生物来源未提及]的重组ADI，并评估了其抗癌活性。该酶在人类生理条件（pH 7和37°C）下稳定，米氏常数为1.90 mM。发现PfADI能有效抑制HepG2细胞，IC50值为0.1950 IU/ml。因此，当前的计算机模拟和体外研究确定PfADI是一种具有提高疗效和低免疫原性的潜在抗癌药物候选物。
在线版本包含可在10.1007/s13205-022-03292-2获取的补充材料。
