Decreasing translation error rate in Escherichia coli increases protein function. Issue 1 (December 2016)

• Record Type: Journal Article
• Authors: Musa, Marina; Radman, Miroslav; Krisko, Anita
• Is Part Of: BMC biotechnology. Volume 16:Issue 1(2016)
• Page Start: 1
• Page End: 5
• Publication Date: 2016-12
• Subjects: Protein activity -- rpsL141 -- Protein expression systems; Biotechnology -- Periodicals; 660.605
• DOI: 10.1186/s12896-016-0259-8
• Languages: English
• ISSNs: 1472-6750

Abstract

Abstract Background Over-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 bearingrpsL141 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 fromEscherichia coli bearingrpsL141 mutation have superior activity compared to those purified from wild typeE. coli, as well as some commercially available industrial enzymes. Results Our results indicate that both alpha-glucosidase and beta-glucanase isolated fromE. coli bearingrpsL141 mutation have increased activity compared to those isolated from wild typeE. coli . Alpha-glucosidase fromrpsL141 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. Conclusion Reduction of the error rate in protein biosynthesis via ribosomalrpsL141 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.