Structural conversion from non-native to native form of recombinant human epidermal growth factor by Brevibacillus choshinensis.
Brevibacillus choshinensis (Bacillus brevis) HPD31 is a very efficient producer of recombinant human epidermal growth factor (EGF). The produced EGF is secreted into the medium with high efficiency. However part of the EGF that accumulates in the medium, exists as multimeric forms which are biologically inactive. We found the bacterium has the activity to structurally convert multimeric forms to the monomeric, native ones. Optimal temperature and pH for the conversion were 40 degrees C and pH 9, respectively. The reaction was promoted in the presence of reduced glutathione or cysteine. But the cells which had been sonicated or exposed to moderate heat treatment completely lost the activity. Thus, it was presumed that the activity might be due to the enzyme(s) that catalyze the protein disulfide exchanging reaction, and that they resides on the surface of viable cells.
Miyauchi
A
A
R & D Department, Higeta Shoyu Co., Ltd., Chiba, Japan. a-miyauchi@higeta.co.jp
Ozawa
M
M
Mizukami
M
M
Yashiro
K
K
Ebisu
S
S
Tojo
T
T
Fujii
T
T
Takagi
H
H
D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D003545:Cysteine / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046911:Macromolecular Substances; D011490:Protein Disulfide Reductase (Glutathione) / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013816:Thermodynamics
D046911:Macromolecular Substances; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor; D011490:Protein Disulfide Reductase (Glutathione); D005978:Glutathione; D003545:Cysteine