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. 2022 Jul 7;10(7):1373.
doi: 10.3390/microorganisms10071373.

Construction of HGF-Displaying Yeast by Cell Surface Engineering

Seiji Shibasaki  1   2 , Yuki Nakatani  3 , Kazuaki Taketani  3 , Miki Karasaki  2   4 , Kiyoshi Matsui  2 , Mitsuyoshi Ueda  4 , Tsuyoshi Iwasaki  2   3
Affiliations

Construction of HGF-Displaying Yeast by Cell Surface Engineering

Seiji Shibasaki et al. Microorganisms. .

Abstract

Hepatocyte growth factor (HGF) has been investigated as a regulator for immune reactions caused by transplantation and autoimmune diseases and other biological functions. Previous studies demonstrated that cDNA-encoding HGF administration could inhibit acute graft-versus-host disease (GVHD) after treatment via hematopoietic stem cell transplantation. This study aimed to show the preparation of HGF protein on yeast cell surfaces to develop a tool for the oral administration of HGF to a GVHD mouse model. In this study, full-length HGF and the heavy chain of HGF were genetically fused with α-agglutinin and were successfully displayed on the yeast cell surface. This study suggested that yeast cell surface display engineering could provide a novel administration route for HGF.

Keywords: cell surface; graft versus host disease; hepatocyte growth factor-displaying; oral administration; yeast cells.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Plasmid constructs for HGF display on the yeast cell surface. (A) Genetic fusion of HGF and α-agglutinin. Leu2-d in the plasmid is inserted for a multiple copy in the yeast cell. (B) Schematics of HGF production on the yeast cell surface. HGF, hepatocyte growth factor.
Figure 2
Figure 2
Immunofluorescence microscopic observation of HGF on Saccharomyces cerevisiae. (A–C) Light field micrograph. (D–F) Fluorescence micrograph. (A,D) BY4741/HGF-FL; (B,E), BY4741/HGF-HC; (C,F), BY4741/pULD1 (control). Dashed circles in (D) indicate areas with fluorescent cells. HGF, hepatocyte growth factor. Scale bar = 5 μm.
Figure 3
Figure 3
Evaluation of HGF-displaying cells during culture. (A) Cell growth. Circle, control; square, BY4741/HGF-FL (full length), diamond, BY4741/HGF-HC (heavy chain). (B) The fluorescence intensity from the yeast cell surface was measured and expressed in relative fluorescence units (RFU). Gray, BY4741/HGF-FL; White, BY4741/HGF-HC. Data represent the means ± SD of three independent experiments. SD, standard deviation; HGF, hepatocyte growth factor.
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