Critical points worthy of consideration in the soluble expression of
recombinant proteins in Escherichia coli: the accuracy of the solubility
prediction tools versus experimental results
Abstract
Abstract Purpose: Recombinant proteins have become increasingly
important items in research and industry. Due to its low cost, high
yield and rapid growth rate, Escherichia coli (E. coli) is the first
choice as host for the production of recombinant proteins. The
expression of recombinant proteins in E. coli systems often result in
inclusion bodies lacking proper folding and structure. In silico
bioinformatics prediction tools may be promising in optimal expression
of soluble recombinant proteins. Materials and methods: In this review,
we aimed at making critical recommendations on how to improve the
soluble expression of recombinant proteins. Furthermore, we compared the
solubility of recombinant proteins using bioinformatics prediction tools
versus experimental results. Data were analyzed using SPSS software.
Results: Some recommendations worthy of consideration in gene design and
expression were reminded. The results of a comparison between
bioinformatics and experimental methods revealed that no significant
coordination existed. RPSP and SOLpro showed higher sensitivity (43.5%
and 56.5%, respectively) and specificity (52.9% and 47.1%,
respectively), when compared to FoldIndex and PSoL. The results from
p-value and roc curve indicated the effect of MW, helix percentage and
aliphatic index on protein solubility (p-value< 0.05).
Conclusions: This review discusses efficient expression of soluble
recombinant proteins. The bioinformatics prediction tools were examined
for their sensitivity and specificity. MW, helix percentage and
aliphatic parameters should be considered in gene design.