Background: Arginine deiminase of Mycoplasma hominis, an arginine catabolizing enzyme, is currently in clinical trial for the treatment of arginine auxotrophic cancers. However, some drawbacks such as instability and antigenicity have limited its application as a protein drug. Arginine Deiminase (ADI) belongs to the guanidino-group modifying enzyme superfamily. Despite differences in the primary amino acid sequences of various members of this superfamily, the folding and secondary structures are conserved in all members. Despite structural similarities, ADIs in various species have different levels of catalytic activity and physicochemical properties due to the differences in their primary amino acid sequences. Therefore, investigating and comparing sequences between different ADI producing bacterial strains could be helpful in the rational engineering of ADI.

Objective: In the current research, we used an in-silico approach to characterize and classify the available reviewed protein sequences of ADI.

Results: 102 ADI sequences from SwissProt database were extracted. Subsequently, based on clustering analyses, the sequence sets were divided into five distinct groups. Different physicochemical properties, solubility, and antigenicity of the enzymes were determined. Some ADI sequences were introduced as well-suited candidates for protein engineering; Lactobacillus fermentum ADI for low pI value, Mycobacterium avium ADI for high aliphatic index, Bacillus licheniformis ADI for low GRAVY index, Bradyrhizobium diazoefficiens ADI for low antigenicity and high stability index, and among Mycoplasma ADIs, Mycoplasma arthritidis ADI for high stability and aliphatic index, and Mycoplasma capricolum for low antigenicity.

背景：人型支原体的精氨酸脱亚氨酶，一种精氨酸分解代谢酶，目前正在临床试验中，用于治疗精氨酸营养缺陷型癌症。但是，诸如不稳定性和抗原性之类的一些缺点限制了其作为蛋白质药物的应用。精氨酸脱亚氨酶（ADI）属于胍基修饰酶超家族。尽管该超家族的各个成员的一级氨基酸序列不同，但折叠和二级结构在所有成员中均是保守的。尽管结构相似，但由于其一级氨基酸序列的差异，各种物种的ADI具有不同水平的催化活性和理化性质。因此，研究和比较产生ADI的不同细菌菌株之间的序列可能有助于ADI的合理设计。

目的：在当前的研究中，我们使用了计算机模拟方法来表征和分类ADI的可用综述蛋白序列。

结果：从SwissProt数据库中提取了102条ADI序列。随后，基于聚类分析，将序列集分为五个不同的组。确定了酶的不同物理化学性质，溶解度和抗原性。介绍了一些ADI序列作为蛋白质工程的合适候选物；发酵乳杆菌ADI用于低pI值，鸟分枝杆菌ADI用于高脂肪指数，地衣芽孢杆菌ADI用于低GRAVY指数，重氮根瘤菌缓释剂ADI用于低抗原性和高稳定性指数，支原体ADIs，支原体关节炎， ADI用于高稳定性和脂肪族指数和毛状支原体的抗原性低。