Bacterial Surface Display of a Co-Factor Containing Enzyme, ω-Transaminase from<i>Vibrio fluvialis</i>Using the<i>Bacillus subtilis</i>Spore Display System

Hwang, Bum-Yeol; Kim, Byung‐Gee; Kim, June-Hyung

doi:10.1271/bbb.110307

Cited by 23 publications

(14 citation statements)

References 18 publications

(17 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Hence, with the absence of caddy protein, cloning and expression of single gene, BMT, is much easier and simple way for the functional surface display compared to ST. Furthermore, the spore display system is very suitable for the surface expression of multimeric proteins such as, streptavidin and ω ‐transaminase and β ‐galactosidase .…”

Section: Discussionmentioning

confidence: 99%

Surface display of bacterial tyrosinase on spores of Bacillus subtilis using CotE as an anchor protein

Hosseini-Abari

Kim

Lee

et al. 2016

J. Basic Microbiol.

Self Cite

View full text Add to dashboard Cite

Tyrosinases, copper-containing monooxygenases, are widely used enzymes for industrial, medical, and environmental applications. We report the first functional surface display of Bacillus megaterium tyrosinase on Bacillus subtilis spores using CotE as an anchor protein. Flow Cytometry was used to verify surface expression of tyrosinase on the purified spores. Moreover, tyrosinase activity of the displayed enzyme on B. subtilis spores was monitored in the presence of L-tyrosine (substrate) and CuSO (inducer). The stability of the spore-displayed tyrosinase was then evaluated after 15 days maintenance of the spores at room temperature, and no significant decrease in the enzyme activity was observed. In addition, the tyrosinase-expressing spores could be repeatedly used with 62% retained enzymatic activity after six times washing with Tris-HCl buffer. This genetically immobilized tyrosinase on the spores would make a new advance in industrial, medical, and environmental applications.

show abstract

Section: Discussionmentioning

confidence: 99%

Surface display of bacterial tyrosinase on spores of Bacillus subtilis using CotE as an anchor protein

Hosseini-Abari

Kim

Lee

et al. 2016

J. Basic Microbiol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Various authors have tested the employment of spores as particles for surface display suitable for vaccination [9-12,19], for review see [5-8,13]. In this work we continue the use of subunit A of the urease (UreA) of Helicobacter acinonychis [10], which has been used extensively as an antigen able to induce, an immune response [20-24].…”

Section: Discussionmentioning

confidence: 99%

New stable anchor protein and peptide linker suitable for successful spore surface display in B. subtilis

2013

View full text Add to dashboard Cite

BackgroundIn last decade spores have been successfully used as a surface display platform. Various peptides or proteins were displayed this way as functional enzymes or antigens. Nearly all attempts involved use of three coat proteins: CotB, CotC or CotG. Increasing knowledge of the structure of the spore coat allowed us to propose the use of other proteins whose localization in the spore envelope has been determined. We also propose the application of a new linker suitable for building fusion proteins.ResultsWe show that a member of the outer coat, CotZ, is a good candidate as a new anchor protein useful in spore surface display. This protein allows use of relatively large passenger proteins and their efficient display on the spore surface. Analysis by Western- and dot-blotting, combined with immunofluorescence microscopy, allowed us to estimate the number of displayed fusion proteins molecules as 1.4 × 102 per spore. In addition, we present data indicating that the use of a peptide linker, which forms a stable α-helix, may greatly improve the display of anchored proteins on the spore surface.ConclusionCotZ can be used as an efficient anchor protein in the outer spore coat. Its localisation in the coat crust layer should guarantee surface display of passenger proteins. Moreover, a CotZ based fusion can tolerate relatively large passenger proteins for efficient spore surface display. In addition, to the properties of both the anchor and passenger proteins, an important issue is the nature of the linker. Here we present evidence that the linker, which forms a stable α-helix, may be crucial for successful display.

show abstract

“…Spore surface display technology can successfully express large molecular weight and multimeric exogenous proteins on the surfaces of spores without crossing the cell membrane. For example, Hwang et al [41] used spore surface display technology to produce pyridoxal phosphate (PLP) in the presence of CotG, used as an anchoring motif, to display dimers on the surfaces of B. subtilis spores. The enzymatic activity of the fusion protein was more than 30 times greater than that of the host spores.…”

Section: Production Of Macromolecular Multimeric Proteinsmentioning

confidence: 99%

Bacillus subtilis Spore Surface Display Technology: A Review of Its Development and Applications

Zhang

Zabed

et al. 2019

Journal of Microbiology and Biotechnology

View full text Add to dashboard Cite

Bacillus subtilis spore surface display (BSSD) technology is considered to be one of the most promising approaches for expressing heterologous proteins with high activity and stability. Currently, this technology is used for various purposes, such as the production of enzymes, oral vaccines, drugs and multimeric proteins, and the control of environmental pollution. This paper presents an overview of the latest developments in BSSD technology and its application in protein engineering. Finally, the major limitations of this technology and future directions for its research are discussed.

show abstract

Bacterial Surface Display of a Co-Factor Containing Enzyme, ω-Transaminase fromVibrio fluvialisUsing theBacillus subtilisSpore Display System

Cited by 23 publications

References 18 publications

Surface display of bacterial tyrosinase on spores of Bacillus subtilis using CotE as an anchor protein

Surface display of bacterial tyrosinase on spores of Bacillus subtilis using CotE as an anchor protein

New stable anchor protein and peptide linker suitable for successful spore surface display in B. subtilis

Bacillus subtilis Spore Surface Display Technology: A Review of Its Development and Applications

Contact Info

Product

Resources

About