Plasmid Display for Stabilization of Enzymes Inside the Cell to Improve Whole-Cell Biotransformation Efficiency

Yunjeong Park, Jonghyeok Shin, Jinkyeong Yang, Hooyeon Kim, Younghun Jung, Hyunseok Oh, Yongjoon Kim, Jaehyeon Hwang, Myeongseo Park, Choongjin Ban, Ki Jun Jeong, Sun Ki Kim, Dae Hyuk Kweon

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Recombinant whole-cell biocatalysts are widely used for biotransformation of valuable products. However, some key enzymes involved in biotransformation processes are unstable and cannot be easily expressed in the functional form. In this study, we describe a versatile platform for enzyme stabilization inside the cell: Intracellularly Immobilized Enzyme System (IIES). A 1,2-fucosyltransferase from Pedobactor saltans (PsFL) and a 1,3-fucosyltransferase from Helicobacter pylori (HpFL), chosen as model proteins, were fused with Oct-1 DNA-binding domain, which mediated the formation of a plasmid–protein complex. Oct-1 fusion enabled both soluble and stable expression of recombinant proteins in the cytoplasm because the fusion proteins were stabilized on the plasmid like immobilized enzymes bound to solid surface. As a result, Oct-1-fusion proteins exhibited significantly greater product titer and yield than non-fusion proteins. Use of fusion proteins PsFL-Oct-1 with C-terminal Oct-1 and Oct-1-PsFL with N-terminal Oct-1 resulted in ~3- and ~2-fold higher 2′-fucosyllactose titers, respectively, than with the use of PsFL alone. When Oct-1 was fused to HpFL, which requires dimerization through heptad repeats, almost two times more 3-fucosyllactose was produced. Fucosyllactose has been used as a food additive because it has various beneficial effects on human health. We anticipate that IIES using Oct-1 fusion protein developed in this study can be applied to stabilize other unstable enzymes.

Original languageEnglish
Article number444
JournalFrontiers in Bioengineering and Biotechnology
Volume7
DOIs
StatePublished - 10 Jan 2020

Keywords

  • DNA binding protein
  • Escherichia coli
  • Oct-1
  • intracellularly immobilized enzyme system
  • whole-cell biotransformation

Fingerprint

Dive into the research topics of 'Plasmid Display for Stabilization of Enzymes Inside the Cell to Improve Whole-Cell Biotransformation Efficiency'. Together they form a unique fingerprint.

Cite this