Premium
Tolerance of Escherichia coli β‐galactosidase C‐terminus to different‐sized fusions
Author(s) -
Corchero José L.,
Villaverde Antonio
Publication year - 1999
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/(sici)1097-0290(19990920)64:6<644::aid-bit2>3.0.co;2-#
Subject(s) - proteolysis , protease , recombinant dna , fusion protein , c terminus , chemistry , biochemistry , biology , microbiology and biotechnology , enzyme , gene , amino acid
The tolerance of the β‐galactosidase C‐terminus to foreign protein fusions has been explored by using different‐sized derivatives of the chimeric protein LACVP1. While the molecular mass of the partner domain shows a minor influence on protein toxicity for the producing E. coli cells, it dramatically affects the proteolytic susceptibility of the whole fusion. Surprisingly, the observed structural modulation of proteolysis is not an all‐or‐nothing process, but it exhibits a continuous effect concomitantly with the length of the fusion. The conformational effects caused by increasingly sized partners seem to progressively expose cryptic protease target sites, initiating a proteolytic cascade that dramatically reduces the yield of the recombinant protein. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 64: 644–649, 1999.