3′-UTR engineering to improve soluble expression and fine-tuning of activity of cascade enzymes in Escherichia coli
SCIE
SCOPUS
- Title
- 3′-UTR engineering to improve soluble expression and fine-tuning of activity of cascade enzymes in Escherichia coli
- Authors
- Song, JW; Woo, JM; Jung, GY; Bornscheuer, UT; Park, JB
- Date Issued
- 2016-07-11
- Publisher
- Nature Publishing Group
- 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.
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/39254
- DOI
- 10.1038/SREP29406
- ISSN
- 2045-2322
- Article Type
- Article
- Citation
- Scientific Reports, vol. 6, 2016-07-11
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