Heterologous Expression of Cyanobacterial Cyanase Gene (CYN) in Microalga Chlamydomonas reinhardtii for Bioremediation of Cyanide Pollution

SIMPLE SUMMARY: Cyanide is a known toxic compound produced through natural and anthropogenic activities. Water can be polluted by cyanide ions through wastewater effluents. In high concentrations, cyanide is considered a strong metabolic inhibitor and can cause inhibition in mitochondrial complex IV (cytochrome c oxidase), and its assimilation can result in chronic poisoning and/or acute poisoning to humans and animals. Bioremediation systems involving the usage of transgenic algal systems have become preferable alternatives for the detoxification of cyanide contamination due to the accumulation and the biosorbent efficiency of transgenic Chlamydomonas in the removal of KCNO from fresh water. ABSTRACT: Recombinant DNA technology offered the creation of new combinations of DNA segments that are not found together in nature. The present study aimed to produce an ecofriendly bioremediation model to remediate cyanide pollution from a polluted marine system. Cyanide is a known toxic compound produced through natural and anthropogenic activities. An Agrobacterium-tumefaciens-mediated genetic transformation technique was used to generate transformed Chlamydomonas reinhardtii using plant expression vector pTRA-K-cTp carries isolated coding sequence of the cyanobacterial cyanase gene (CYN) isolated from Synechococcus elongatus (PCC6803). qRT-PCR analysis showed the overexpression of CYN in transgenic C. reinhardtii, as compared with the respective wild type. Growth parameters and biochemical analyses were performed under cyanide stress conditions using transgenic and wild C. reinhardtii for evaluating the effect of the presence of the cyanobacterial cyanase gene in algae. The transgenic C. reinhardtii strain (TC. reinhardtii-2) showed promising results for cyanide bioremediation in polluted water samples. Cyanide depletion assays and algal growth showed a significant resistance in the transgenic type against cyanide stress, as compared to the wild type. Genetically modified alga showed the ability to phytoremediate a high level of potassium cyanide (up to150 mg/L), as compared to the wild type. The presence of the CYN gene has induced a protection response in TC. Reinhardtii-2, which was shown in the results of growth parameter analyses. Therefore, the present study affirms that transgenic C. reinhardtii by the CYN coding gene is a potential effective ecofriendly bioremediator model for the remediation of cyanide pollutants in fresh water.