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首页> 外文期刊>Applied Microbiology and Biotechnology >Carboxylesterase 2 production and characterization in human cells: New insights into enzyme oligomerization and activity
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Carboxylesterase 2 production and characterization in human cells: New insights into enzyme oligomerization and activity

机译:人体细胞中羧酯酶2的产生和表征:酶寡聚和活性的新见解

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摘要

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 10× 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.
机译:羧基酯酶2(CES2)是人肠中表达的主要羧酸酯酶,在抗癌联合疗法中用于治疗结肠癌和恶性神经胶质瘤等不同病理学方面的酶越来越重要。本文描述了使用无血清培养基在人胚胎肾细胞(HEK-293T细胞)中产生人重组CES2。通过简单地添加框内C端10x组氨酸标签(CES2-10xHis)即可实现CES2向培养基的分泌,而无需添加额外的N端信号序列或C端突变或缺失HTEL基序负责将蛋白质保留在内质网腔中。这种分泌使CES2产量增加了四倍。人类重组CES2的特征表明,该蛋白质以不同于所述60 kDa单体的其他活性和非活性形式存在。

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