Modified penicillin acylase signal peptide allows the periplasmic production of soluble human interferon-gamma but not of soluble human interleukin-2 by the Tat pathway in Escherichia coli.
Production of periplasmic human interferon-gamma (hINF-gamma) and human interleukin-2 (hIL-2) by the Tat translocation pathway in Escherichia coli BL21-SI was evaluated. The expression was obtained using the pEMR vector which contains the Tat-dependent modified penicillin acylase signal peptide (mSPpac) driven by the T7 promoter. The mSPpac-hINF-gamma was processed and the protein was transported to periplasm. Up to 30.1% of hINF-gamma was found in the periplasmic soluble fraction, whereas only 15% of the mSPpac-hIL-2 was processed, but hIL-2 was not found in the periplasmic soluble fraction.
Medina-Rivero
E
E
Division of Molecular Biology, Institute for Scientific and Technological Research of San Luis Potosi, Camino a la Presa San Jose 2055 Lomas 4a. Sección, C.P. 78216, San Luis Potosi, SLP, Mexico.
Balderas-Hernández
V E
VE
Ordoñez-Acevedo
L G
LG
Paz-Maldonado
L M T
LM
Barba-De la Rosa
A P
AP
De León-Rodríguez
A
A
D004926:Escherichia coli / Q000502:physiology*; D006801:Humans; D007371:Interferon-gamma / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007376:Interleukin-2 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019897:Periplasm / Q000737:chemistry; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism; D012995:Solubility
D007376:Interleukin-2; D011994:Recombinant Proteins; D007371:Interferon-gamma