Stabilization of proteins inside cells by immobilization on plasmid

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Article Title

Stabilization of proteins inside cells by immobilization on plasmid

Year

2019

Publication Date

8/1/2019

Authors

PARK, Yunjeong

Data Format

PDF

Delivery Time (Days)

1

Abstract

Enzymes perform a huge diversity of reactions in living cells. It is important to
express enzymes in a soluble form to maintain their activity inside cells. When soluble
expression of enzymes involved in the metabolic pathway leading to product formation
is problematic, various methods can be applied to improve the solubility of the enzyme.
Fusion with soluble tags such as Glutathione S-transferases (GSTs), Maltose-binding
protein (MBP), and thioredoxin is the most commonly employed method. However, all
proteins are not expressed soluble by these fusion tags. In this study, we describe a
versatile platform for protein stabilization inside the cell, called Intracellular
Immobilization Enzyme System (IIES). To design this IIES system, we fused target
protein to a DNA binding domain. IIES helps to form plasmid-protein complex by the
help of DNA binding protein and binding sequence on the plasmid. Plasmids are known
to be one of the stable substances in cells. When the plasmid was used as a carrier, the
enzyme could be stabilized in the cytoplasm like an immobilized enzyme bound to solid
surface. When IIES was applied to make a whole cell biocatalyst through metabolic
engineering, such fusion proteins substantially increased productivity. Therefore, the plasmid-protein complex forms an immobilized enzyme and the plasmid construct acts
as an immobilization carrier, and the use of intracellular immobilized enzymes is likely
to be particularly useful for metabolic engineering.

Keywords

solubility tag, DNA binding protein, immobilized enzyme, protein
stabilization, biotransformation

Author Affiliation/Institution

Sungkyunkwan University

Subject Fields

Biology