I am trying to express HIV1 antisense protein in E.Coli. The protein is hydrophobic and also has two N-terminal cysteine triplets. I am planning to add a pelb signal peptide for periplasmic secretion. will it work or I should go with thioredoxin fusion partner?
Dear Balakumaran Sathiyamani
Indeed, (in general) E. coli recombinant protein production systems are designed to achieve a high accumulation of soluble protein product in the cytosol. However, in the following excellent review:
Rosano, G. L., & Ceccarelli, E. A. (2014). Recombinant protein expression in Escherichia coli: advances and challenges. Frontiers in microbiology, 5, 172. https://www.frontiersin.org/articles/10.3389/fmicb.2014.00172/full
they rightfully state and I quote “Secretion to the periplasm or to the medium is sometimes the only way to produce a recombinant protein (Mergulhao et al., 2005; de Marco, 2009)… The signal peptides of the following proteins are widely used for secretion: Lpp, LamB, LTB, MalE, OmpA, OmpC, OmpF, OmpT, PelB, PhoA, PhoE, or SpA (Choi and Lee, 2004).” So, yes using PelB is worthwhile trying.
However, keep in mind that the selection the appropriate signal sequence is just one of the factors that determines the level of expression, see for example:
Jashandeep Kaur, Arbind Kumar, Jagdeep Kaur (2018) Strategies for optimization of heterologous protein expression in E. coli: Roadblocks and reinforcements. Int. J. Biol. Macromol. 106, 803-822 https://doi.org/10.1016/j.ijbiomac.2017.08.080
Best regards.
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