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Functional display of foreign protein on surface of Escherichia coli using N‐terminal domain of ice nucleation protein
Author(s) -
Li Lin,
Gyun Kang Dong,
Joon Cha Hyung
Publication year - 2003
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10892
Subject(s) - fusion protein , escherichia coli , green fluorescent protein , chemistry , cell , microbiology and biotechnology , biochemistry , biology , gene , recombinant dna
We investigated the ability of the N‐terminal domain of InaK, an ice nucleation protein from Pseudomonas syringae KCTC1832, to act as an anchoring motif for the display of foreign proteins on the Escherichia coli cell surface. Total expression level and surface display efficiency of green fluorescent protein (GFP) was compared following their fusion with either the N‐terminal domain of InaK (InaK‐N), or with the known truncated InaK containing both N‐ and C‐terminal domains (InaK‐NC). We report that the InaK‐N/GFP fusion protein showed a similar cell surface display efficiency (∼50%) as InaK‐NC/GFP, demonstrating that the InaK N‐terminal region alone can direct translocation of foreign proteins to the cell surface and can be employed as a potential cell surface display motif. Moreover, InaK‐N/GFP showed the highest levels of total expression and surface display based on unit cell density. InaK‐N was also successful in directing cell surface display of organophosphorus hydrolase (OPH), confirming its ability to act as a display motif. © 2003 Wiley Periodicals, Inc.