Azurin, a bacteriocin produced by a human gut bacterium Pseudomonas aeruginosa, can reveal selectively cytotoxic and induce apoptosis in cancer cells. After overcoming two phase I trials, a functional region of Azurin called p28 has been approved as a drug for the treatment of brain tumor glioma by FDA. The present study aims to improve a screening procedure and assess genetic diversity of Azurin genes in P. aeruginosa and Azurin-like genes in the gut microbiome of a specific population in Vietnam and global populations. Firstly, both cultivation-dependent and cultivation-independent techniques based on genomic and metagenomic DNAs extracted from fecal samples of the healthy specific population were performed and optimized to detect Azurin genes. Secondly, the Azurin gene sequences were analyzed and compared with global populations by using bioinformatics tools. Finally, the screening procedure improved from the first step was applied for screening Azurin-like genes, followed by the protein synthesis and NCI in vitro screening for anticancer activity. As a result, this study has successfully optimized the annealing temperatures to amplify DNAs for screening Azurin genes and applying to Azurin-like genes from human gut microbiota. The novelty of this study is the first of its kind to classify Azurin genes into five different genotypes at a global scale and confirm the potential anticancer activity of three Azurin-like synthetic proteins (Cnazu1, Dlazu11, and Ruazu12). The results contribute to the procedure development applied for screening anticancer proteins from human microbiome and a comprehensive understanding of their therapeutic response at a genetic level.

中文翻译：

---

通过依赖于培养和不依赖培养的研究推导的人类肠道微生物组中抗癌Azurin编码和Azurin样基因的分子筛选和遗传多样性分析。

天青素是人的肠道细菌铜绿假单胞菌产生的一种细菌素，可以选择性地显示出细胞毒性并诱导癌细胞凋亡。克服了两项I期试验后，FDA批准了称为p28的Azurin功能区被批准为治疗脑肿瘤神经胶质瘤的药物。本研究旨在改善筛选程序并评估铜绿假单胞菌中天青素基因的遗传多样性越南特定人群和全球人群的肠道微生物组中的Azurin样基因。首先，基于从健康特定人群的粪便样本中提取的基因组和宏基因组DNA的依赖于培养和不依赖于培养的技术都进行了优化，以检测Azurin基因。其次，使用生物信息学工具分析了Azurin基因序列，并将其与全球人群进行了比较。最后，从第一步开始改进的筛选程序用于筛选类似Azurin的基因，然后进行蛋白质合成和NCI体外筛选抗癌活性。结果，该研究成功地优化了退火温度，以扩增用于筛选天青蛋白基因的DNA，并将其应用于人类肠道菌群中的类似天青蛋白的基因。这项研究的新颖性是首次在全球范围内将天青素基因分类为五种不同基因型，并证实了三种天青素样合成蛋白（Cnazu1，Dlazu11和Ruazu12）的潜在抗癌活性。该结果有助于从人类微生物组中筛选抗癌蛋白的程序开发，并全面了解其在基因水平上的治疗反应。