Decreasing translation error rate in Escherichia coli increases protein functionReport as inadecuate

Decreasing translation error rate in Escherichia coli increases protein function - Download this document for free, or read online. Document in PDF available to download.

BMC Biotechnology

, 16:28

Protein and enzyme technology


BackgroundOver-expressed native or recombinant proteins are commonly used for industrial and pharmaceutical purposes, as well as for research. Proteins of interest need to be purified in sufficient quantity, quality and specific activity to justify their commercial price and eventual medical use. Proteome quality was previously positively correlated with ribosomal fidelity, but not on a single protein level. Here, we show that decreasing translational error rate increases the activity of single proteins. In order to decrease the amount of enzyme needed for catalysis, we propose an expression system bearing rpsL141 mutation, which confers high ribosomal fidelity. Using alpha-glucosidase exo-alpha-1,4-glucosidase and beta-glucanase beta-D-glucanase as examples, we show that proteins purified from Escherichia coli bearing rpsL141 mutation have superior activity compared to those purified from wild type E. coli, as well as some commercially available industrial enzymes.

ResultsOur results indicate that both alpha-glucosidase and beta-glucanase isolated from E. coli bearing rpsL141 mutation have increased activity compared to those isolated from wild type E. coli. Alpha-glucosidase from rpsL141 background has a higher activity than the purchased enzymes, while beta-glucanase from the same background has a higher activity compared to the beta-glucanase purchased from Sigma, but not compared to the one purchased from Megazyme.

ConclusionReduction of the error rate in protein biosynthesis via ribosomal rpsL141 mutation results in superior functionality of single proteins. We conclude that this is a viable system for expressing proteins with higher activity and that it can be easily scaled up and combined with other expression systems to meet the industrial needs.

KeywordsProtein activity rpsL141 Protein expression systems  Download fulltext PDF

Author: Marina Musa - Miroslav Radman - Anita Krisko


Related documents