R. Yatsunami, M. Sato, K. Orishimo, Y. Hatori, Y. Zhang, T. Takashina, T. Fukui, Satoshi Nakamura
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引用次数: 9
摘要
在极端嗜盐古细菌盐盐菌NRC-1基因组中发现了一个编码几丁质酶同源物(ChiN1)的开放阅读框。ChiN1是一种多结构域酶,由几丁质结合结构域、多囊肾病结构域和催化结构域组成,属于糖苷水解酶家族18。利用细胞表面糖蛋白基因启动子序列成功表达了chiN1基因在极嗜盐古菌(Haloarcula japonica TR-1)中的表达。大量重组ChiN1被分泌到培养上清液中。Ha。对日本产的ChiN1进行了纯化和鉴定。ChiN1的最佳pH和温度分别为pH 4.5和55℃。ChiN1在1.0 M NaCl条件下活性最强,在1.0 ~ 4.5 M NaCl浓度范围内稳定。这是对极嗜盐古菌中几丁质酶的首次报道。
Gene expression and characterization of a novel GH family 18 chitinase from extremely halophilic archaeon Halobacterium salinarum NRC-1
An open reading frame encoding a chitinase homolog (ChiN1) was found in the genome of extremely halophilic archaeon Halobacterium salinarum NRC-1. ChiN1 is a multidomain enzyme consisting of a chitin-binding domain, a polycystic kidney disease domain and a catalytic domain belonging to glycoside hydrolase family 18. chiN1 gene was successfully expressed in extremely halophilic archaeon Haloarcula japonica TR-1 by employing the promoter sequence of its cell surface glycoprotein gene. A large amount of recombinant ChiN1 was secreted into the culture supernatant. The Ha. japonica-produced ChiN1 was purified and characterized. The optimal pH and temperature of ChiN1 are pH 4.5 and 55°C, respectively. ChiN1 was most active at 1.0 M NaCl and stable over a wide range of NaCl concentration from 1.0 to 4.5 M. This is the first report on a chitinase from extremely halophilic archaeon.