Optimisation of signal peptide for recombinant protein secretion in bacterial hosts

Abstract

Escherichia coli-the powerhouse for recombinant protein production-is rapidly gaining status as a reliable and efficient host for secretory expression. An improved understanding of protein translocation processes and its mechanisms has inspired and accelerated the development of new tools and applications in this field and, in particular, a more efficient secretion signal. Several important characteristics and requirements are summarised for the design of a more efficient signal peptide for the production of recombinant proteins in E. coli. General approaches and strategies to optimise the signal peptide, including the selection and modification of the signal peptide components, are included. Several challenges in the secretory production of recombinant proteins are discussed, and research approaches designed to meet these challenges are proposed.

In Silico Evaluation of Different Signal Peptides for the Secretory Production of Human Growth Hormone in E. coli

“…It has been also believed that the presence of the basic residue, particularly lysine, at the second codon (P2) improves the rate of protein secretion (Puziss et al 1992). Therefore, the presence of one or more basic amino acids in the n-region is probably essential for the development of an applicable signal peptide (Low et al 2013). The net positive charge was 2 for most of the signal peptides in this study, except endo-1.4-betaxylanase and MalE with net positive charges of 4 and 3 respectively, and PelB, PhoA and L-asparaginase II with net positive charge of 1.…”

“…Sec-dependent system is the predominant pathway for protein secretion in E. coli using the sec-dependent signal peptides (e.g. OmpA, PhoA, MalE or LamB) (Low et al 2013;Rusch and Kendall 2007). In addition to the aforementioned system, signal recognition particle (SRP)-dependent pathway is another system for protein secretion through N-terminal signal peptides (e.g.…”

RECONSTITUTION OF DIFFERENT SIGNAL PEPTIDES FOR ENHANCED THERMOSTABLE ALPHA AMYLASE SECRETION IN Bacillus subtilis

“…SPs play a very important role in the transport of secretion proteins by the Sec pathway because they interact with SecA proteins, the signal recognition particle (SRP) and they have recognized site for signal peptidases [5]. The interaction between SP and mature protein is also known to affect the secretion of proteins [5,9]. Therefore, an effective signal peptide for the secretion of any target protein is the most important choice.…”

“…Signal peptide in Sec pathway usually is 18 -40 amino acids long and although the primary structures of different signal peptides show a little similarity and don't have sequence homologous, this signal always consists of three identifiable domains as: a positively charged amino terminal (N-), a central hydrophobic (H-), and carboxyl-terminal (C-) regions [7,8]. N region is 1-5 amino acids long, charged positive, and often has two amino acids Lys and Arg [6,9]. The hydrophobic core (H region) with 7 -15 amino acids takes an alpha helical conformation when it contacts with the membrane lipid phase.…”

Untitled

“…B. lehensis G1 genome contain 4017 protein-coding sequences with approximately 70% assigned biological functions [15]. The availability of this bacterial genome may facilitate researcher to find new potential area to explore, as there are already few articles reported on B. lehensis G1 such as cyclodextrin glucanotransferase [16,17,18], membrane protein [19], and signal peptide [20,21]. In this paper, ongoing works on recombinant proteases isolated from B. lehensis G1 together with the molecular structure prediction will be discussed.…”