TY - JOUR
T1 - Carboxylesterase 2 production and characterization in human cells: new insights into enzyme oligomerization and activity
AU - Coroadinha, Ana Sofia
AU - Simplicio, Ana Luisa
AU - Alves, Paula Maria
PY - 2013/1/1
Y1 - 2013/1/1
N2 - Carboxylesterase 2 (CES2), the main carboxylesterase expressed in human intestine, is an increasingly important enzyme in anti-cancer combined therapies for the treatment of different pathologies like colon adenocarcinoma and malignant glioma. The production of human recombinant CES2, in human embryonic kidney cells (HEK-293T cells) using serum-free media, is herein described. CES2 secretion to the media was achieved by the simple addition of an in-frame C-terminal 10x histidine tag (CES2-10xHis) without the need of addition of extra N-terminal signalling sequences or the mutation or deletion of the C-terminal HTEL motif responsible for retaining the protein in the lumen of endoplasmic reticulum. This secretion allowed a fourfold increase in CES2 production. The characterization of human recombinant CES2 showed that this protein exists in other active and inactive forms than the described 60 kDa monomer
AB - Carboxylesterase 2 (CES2), the main carboxylesterase expressed in human intestine, is an increasingly important enzyme in anti-cancer combined therapies for the treatment of different pathologies like colon adenocarcinoma and malignant glioma. The production of human recombinant CES2, in human embryonic kidney cells (HEK-293T cells) using serum-free media, is herein described. CES2 secretion to the media was achieved by the simple addition of an in-frame C-terminal 10x histidine tag (CES2-10xHis) without the need of addition of extra N-terminal signalling sequences or the mutation or deletion of the C-terminal HTEL motif responsible for retaining the protein in the lumen of endoplasmic reticulum. This secretion allowed a fourfold increase in CES2 production. The characterization of human recombinant CES2 showed that this protein exists in other active and inactive forms than the described 60 kDa monomer
KW - Human cells
KW - Protein expression
KW - Enzyme characterization
KW - Carboxylesterases
U2 - 10.1007/s00253-012-3994-3
DO - 10.1007/s00253-012-3994-3
M3 - Article
SN - 0175-7598
VL - 97
SP - 1161
EP - 1173
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
IS - 3
ER -