从凤梨科植物红苞凤梨果实中分离得到的半胱氨酸蛋白酶——华丽凤梨蛋白酶的初步功能表征、克隆及一级序列分析
Preliminary functional characterization, cloning and primary sequence of Fastuosain, a cysteine peptidase isolated from fruits of Bromelia fastuosa.
作者信息
Cabral Hamilton, Leopoldino Andréia M, Tajara Eloiza H, Greene Lewis J, Faça Vitor M, Mateus Rogério P, Ceron Carlos R, de Souza Judice Wagner A, Julianod Luiz, Bonilla-Rodriguez Gustavo O
机构信息
Departamento de Química e Ciências Ambientais,IBILCE-UNESP, State University of São Paulo, São José do Rio Preto SP, Brazil.
出版信息
Protein Pept Lett. 2006;13(1):83-9. doi: 10.2174/092986606774502072.
The present work reports the characterization of Fastuosain, a novel cysteine protease of 25kDa, purified from the unripe fruits of Bromelia fastuosa, a wild South American Bromeliaceae. Proteolytic activity, measured using casein and synthetic substrates, was dependent on the presence of thiol reagents, having maximum activity at pH 7.0. The present work reports cDNA cloning of Fastuosain; cDNA was amplified by PCR using specific primers. The product was 1096pb long. Mature fastuosain has 217 residues, and with the proregion has a total length of 324 residues. Its primary sequence showed high homology with ananain(74%), stem bromelain (66%) and papain (44%).
本研究报道了从南美洲野生凤梨科植物华丽凤梨未成熟果实中纯化得到的一种新型25kDa半胱氨酸蛋白酶——华丽蛋白酶的特性。使用酪蛋白和合成底物测定的蛋白水解活性依赖于硫醇试剂的存在，在pH 7.0时具有最大活性。本研究报道了华丽蛋白酶的cDNA克隆；使用特异性引物通过PCR扩增cDNA。产物长度为1096pb。成熟的华丽蛋白酶有217个残基，加上前区全长为324个残基。其一级序列与菠萝蛋白酶（74%）、茎菠萝蛋白酶（66%）和木瓜蛋白酶（44%）具有高度同源性。
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