Yildiz Melike, Askin Hakan
a Department of Molecular Biology and Genetics, Faculty of Science, Ataturk University , Erzurum , Turkey.
Prep Biochem Biotechnol. 2019;49(8):800-806. doi: 10.1080/10826068.2019.1621894. Epub 2019 Jun 1.
In this study, azurin, a bacteriocin with anticancer property, was produced by food-grade using the Nisin Controlled Gene Expression (NICE) System. In addition, the antibacterial and cytotoxic properties of recombinant azurin in the culture supernatant were also investigated. Azurin gene from was cloned into the pNZ8149 vector and the resulting recombinant DNA was transformed into food grade NZ3900. The expression of azurin protein was induced by the optimum concentration of nisin for 3 h. Inhibition zones for and were observed at 5.0 and 10 mg/mL concentrations of lyophilized supernatants containing azurin, but no inhibition zone at azurin-free lyophilized supernatants. When HUVEC, HT29, HCT116, and MCF7 cell lines were treated with lyophilized culture supernatants with azurin or without azurin, cell viability decreased with increasing concentrations of the supernatant. Furthermore, the supernatants containing azurin showed more anti-proliferative effect than the azurin-free supernatants. This work provides a practicable method to produce recombinant azurin in the food grade strain. As a result, the recombinant strain, producing azurin, can be used in the investigation of food biopreservatives and in the development of a therapeutic probiotic.
在本研究中，使用乳链菌肽控制基因表达（NICE）系统，通过食品级菌株生产了具有抗癌特性的细菌素天青蛋白。此外，还研究了培养上清液中重组天青蛋白的抗菌和细胞毒性特性。将来自[具体来源未提及]的天青蛋白基因克隆到pNZ8149载体中，并将所得重组DNA转化到食品级菌株NZ3900中。用最佳浓度的乳链菌肽诱导天青蛋白表达3小时。在含有天青蛋白的冻干上清液浓度为5.0和10mg/mL时，观察到对[具体菌种未提及]和[具体菌种未提及]的抑菌圈，但在不含天青蛋白的冻干上清液中未观察到抑菌圈。当用含有或不含天青蛋白的冻干培养上清液处理人脐静脉内皮细胞（HUVEC）、人结肠癌细胞系HT29、人结肠癌细胞系HCT116和人乳腺癌细胞系MCF7时，细胞活力随着上清液浓度的增加而降低。此外，含有天青蛋白的上清液比不含天青蛋白的上清液表现出更强的抗增殖作用。这项工作提供了一种在食品级菌株中生产重组天青蛋白的可行方法。因此，产生天青蛋白的重组菌株可用于食品生物防腐剂的研究和治疗性益生菌的开发。
