Development of a bacteria resistant to high osmolarity through the exploration of biological parts

Abstract

Biotechnology has been a field present since ancient times, and over the years, the development of this science has enabled the creation of various new techniques and knowledge, among them synthetic biology. The ability to alter and create genes, genetic circuits, and organisms made possible by synthetic biology has allowed its use in various areas such as medicine, agriculture, and industry. Within this context, an important application would be in projects involving bioeconomy and sustainability. The use of microorganisms in bioprocesses is highly beneficial, as they can transform substances of little value into valuable products. However, the water footprint caused by these processes is unfavorable to the environment, as they consume large amounts of freshwater. An alternative to this issue would be the use of seawater during the processes, as it constitutes the majority of global water resources. However, seawater has a high concentration of salts, which hinders the growth and survival of microorganisms in this environment. Therefore, obtaining a microorganism that can thrive in these conditions is of utmost importance. Based on this, this work aims to obtain a bacterium resistant to high salinity. To achieve this goal, we will initially proceed with the identification, through literature research and database searches, of the biological parts that respond to saline stress. Next, we will move on to the creation of gene circuits, using the cloning of genes and promoters that were previously identified in the pVANT plasmid. The characterization of these elements will be conducted by cultivating them in a high-salinity medium, allowing us to assess the functionality of these components in the model bacteria Escherichia coli DH10B and Pseudomonas putida KT2440.