fulltext.study @t Gmail

Exploring codon context bias for synthetic gene design of a thermostable invertase in Escherichia coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
16981 42628 2015 7 PDF Available
Title
Exploring codon context bias for synthetic gene design of a thermostable invertase in Escherichia coli
Abstract

•Application of codon context fitness for heterologous enzyme expression in E. coli.•Novel design criteria of secondary structure for codon optimization in E. coli.•Increased invertase activity of 40% by codon context optimization over wild type.•Importance of codon context fitness for heterologous protein expression in E. coli.

Various isoforms of invertases from prokaryotes, fungi, and higher plants has been expressed in Escherichia coli, and codon optimisation is a widely-adopted strategy for improvement of heterologous enzyme expression. Successful synthetic gene design for recombinant protein expression can be done by matching its translational elongation rate against heterologous host organisms via codon optimization. Amongst the various design parameters considered for the gene synthesis, codon context bias has been relatively overlooked compared to individual codon usage which is commonly adopted in most of codon optimization tools. In addition, matching the rates of transcription and translation based on secondary structure may lead to enhanced protein folding. In this study, we evaluated codon context fitness as design criterion for improving the expression of thermostable invertase from Thermotoga maritima in Escherichia coli and explored the relevance of secondary structure regions for folding and expression. We designed three coding sequences by using (1) a commercial vendor optimized gene algorithm, (2) codon context for the whole gene, and (3) codon context based on the secondary structure regions. Then, the codon optimized sequences were transformed and expressed in E. coli. From the resultant enzyme activities and protein yield data, codon context fitness proved to have the highest activity as compared to the wild-type control and other criteria while secondary structure-based strategy is comparable to the control. Codon context bias was shown to be a relevant parameter for enhancing enzyme production in Escherichia coli by codon optimization. Thus, we can effectively design synthetic genes within heterologous host organisms using this criterion.

Keywords
Invertase; Fructofuranosidases; Codon context optimization; Secondary structure; Synthetic gene design; Thermotoga maritima
First Page Preview
Exploring codon context bias for synthetic gene design of a thermostable invertase in Escherichia coli
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volumes 75–76, July–August 2015, Pages 57–63
Authors
, , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us