Maleki Fatemeh, Shahpiri Azar
Department of Biotechnology, College of Agriculture, Isfahan University of Technology, 84156-83111, Isfahan, Iran.
Biometals. 2022 Oct;35(5):889-901. doi: 10.1007/s10534-022-00412-6. Epub 2022 Jun 29.
The toxic nature of arsenic has left a trail of disastrous health consequences around the world. Microorganisms have developed various strategies to deal with arsenic. The presence of plasmid and chromosomal ars operons is one of the most important mechanisms for the detoxification of arsenic in bacteria. ArsR is a trans-acting regulatory protein and acts as a repressor on ars operon. The gene encoding ArsR from Corynebacterium glutamicum (CgArsR1) was cloned in expression vectors pET28a. The resulting constructs were transformed into Escherichia coli strains Rosetta (DE3) and Rosetta gami 2. Following the induction with Isopropyl β-D-1-thiogalactopyranoside, the protein His-CgArsR1 was found in the soluble fraction of strain Rg-CgArsR1. For comparison, ArsR from E. coli was also overexpressed in E. coli (strain Rosetta gami 2) as His-EcArsR. A strain containing empty vector pET28a was also used as a control strain. In the medium containing either arsenite (0.5 mM) or arsenate (0.5 mM), the strain Rg-CgArsR1 and Rg-EcArsR were able to accumulate 1200 and 700 µg/g DCW As, respectively. In comparison, the accumulation of As in these strains was 338 and 232 µg/g DCW, respectively. Whereas both strains Rg-CgArsR1 and Rg-EcArsR were able to accumulate higher amounts of As and As with respect to control strain, the accumulation of arsenic in the strain Rg-CgArsR1 was significantly more efficient than strain Rg-EcArsR for removing As and As. Based on the results the gene encoding CgArsR1 is a useful and efficient target gene for the modification of bacteria for bioremediation of arsenic from polluted soil and water.
砷的毒性在全球范围内留下了一系列灾难性的健康后果。微生物已开发出各种应对砷的策略。质粒和染色体上的ars操纵子的存在是细菌中砷解毒的最重要机制之一。ArsR是一种反式作用调节蛋白,对ars操纵子起阻遏作用。将谷氨酸棒杆菌编码ArsR的基因(CgArsR1)克隆到表达载体pET28a中。将所得构建体转化到大肠杆菌菌株Rosetta (DE3) 和Rosetta gami 2中。用异丙基-β-D-硫代半乳糖苷诱导后,在菌株Rg-CgArsR1的可溶部分中发现了蛋白质His-CgArsR1。为作比较,大肠杆菌的ArsR也在大肠杆菌(菌株Rosetta gami 2)中作为His-EcArsR过表达。含有空载体pET28a的菌株也用作对照菌株。在含有亚砷酸盐(0.5 mM)或砷酸盐(0.5 mM)的培养基中,菌株Rg-CgArsR1和Rg-EcArsR分别能够积累1200和700 μg/g干重的砷。相比之下,这些菌株中砷的积累量分别为338和232 μg/g干重。虽然菌株Rg-CgArsR1和Rg-EcArsR相对于对照菌株都能够积累更多的砷,但菌株Rg-CgArsR1在去除砷和砷方面积累砷的效率明显高于菌株Rg-EcArsR。基于这些结果,编码CgArsR1的基因是用于改造细菌以对污染土壤和水中的砷进行生物修复的有用且高效的靶基因。