Random knock-in expression system for high yield production of heterologous protein in Bacillus subtilis

Cited 14 time in scopus
Metadata Downloads
Title
Random knock-in expression system for high yield production of heterologous protein in Bacillus subtilis
Author(s)
Da-Eun Jeong; Younju So; S Y Park; Seung-Hwan Park; Soo Keun Choi
Bibliographic Citation
Journal of Biotechnology, vol. 266, pp. 50-58
Publication Year
2018
Abstract
Chromosome-integrated recombinant protein expression in bacteria has advantages for the stable maintenance of genes without any use of antibiotics during large-scale fermentation. Even though different levels of gene expression were reported, depending upon their chromosomal position in bacterial species, only a limited number of integration sites have been used in B. subtilis. In this study, we randomly integrated the GFP and AprE expression cassettes into the B. subtilis genome to determine integration sites that can produce a high yield of heterologous protein expression. Our mariner transposon-based expression cassette integration system was able to find integration sites, which can produce up to 2.9-fold and 1.5-fold increased expression of intracellular GFP and extracellular AprE, respectively, compared to the common integration site amyE. By analyzing the location of integration sites, we observed an adjacent promoter effect, gene dosage effect, and gene knock-out effect all complexly contributing to the increased level of integrated gene expression. Besides obtaining a high yield of heterologous protein expression, our system can also provide a wide-range of expression to expand the systematic application for steady-state metabolic protein production. ⓒ 2017 Elsevier B.V.
Keyword
Bacillus expression systemChromosomal integration siteMariner-based transposonRandom knock-in
ISSN
0168-1656
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.jbiotec.2017.12.007
Type
Article
Appears in Collections:
Division of Research on National Challenges > Infectious Disease Research Center > 1. Journal Articles
Files in This Item:
  • There are no files associated with this item.


Items in OpenAccess@KRIBB are protected by copyright, with all rights reserved, unless otherwise indicated.