Extracellular expression of recombinant chitosanase by EschericExtracellular expression of recombinant chitosanase by Escherichia coli using Bacillus subtilis CH2 derived chitosasnasemutation ignal peptidehia coli using Bacillus subtilis CH2 derived chitosasnasemutation signal peptide

Abstract

The industrial production of recombinant proteins secreted by Escherichia coli is essential in the biotechnology industry. Owing to its extensive use as a model system, E. coli genetics have been extensively characterized, resulting in the development of numerous tools that aid in chromosomal engineering, gene replication, and gene expression. Recombinant proteins can be targeted to various compartments within E. coli, including the cytoplasm, periplasm, and extracellular. Extracellular expression in E. coli offers several advantages for the production of recombinant proteins, including simplified downstream processing, reduced host-cell contamination, high expression levels, reduced protein degradation, and increased solubility. E. coli has two main secretory pathways, the Sec and Tat pathways. The Sec pathway, which facilitates the transportation of proteins from the cytoplasmic membrane, is the first secretory pathway identified in bacteria. This pathway exhibits the ability to translocate proteins in an unfolded state, thereby facilitating the efficient export of target proteins. In contrast, the Tat pathway has the distinctive ability to transport fully folded proteins across the cytoplasmic membrane. Previous studies confirmed that chitosanase derived from B. subtilis CH2 utilizes a Sec pathway signal peptide to be secreted into the extracellular space, and further characterized the enzyme's properties. In this study, we aimed to develop an approach to enhance the efficiency of chitosanase secretion in E. coli expression systems by utilizing a signal peptide from B. subtilis CH2.