3′-UTR engineering to improve soluble expression and fine-tuning of activity of cascade enzymes in Escherichia coli

Ji Won Song, Ji Min Woo, Gyoo Yeol Jung, Uwe T. Bornscheuer, Jin Byung Park

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

3′-Untranslated region (3′UTR) engineering was investigated to improve solubility of heterologous proteins (e.g., Baeyer-Villiger monooxygenases (BVMOs)) in Escherichia coli. Insertion of gene fragments containing putative RNase E recognition sites into the 3′UTR of the BVMO genes led to the reduction of mRNA levels in E. coli. Importantly, the amounts of soluble BVMOs were remarkably enhanced resulting in a proportional increase of in vivo catalytic activities. Notably, this increase in biocatalytic activity correlated to the number of putative RNase E endonucleolytic cleavage sites in the 3′UTR. For instance, the biotransformation activity of the BVMO BmoF1 (from Pseudomonas fluorescens DSM50106) in E. coli was linear to the number of RNase E cleavage sites in the 3′UTR. In summary, 3′UTR engineering can be used to improve the soluble expression of heterologous enzymes, thereby fine-tuning the enzyme activity in microbial cells.

Original languageEnglish
Article number29406
JournalScientific Reports
Volume6
DOIs
StatePublished - 11 Jul 2016

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