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Saci_1816:一种催化嗜热嗜酸泉古菌中海藻糖降解的海藻糖酶 。

Saci_1816: A Trehalase that Catalyzes Trehalose Degradation in the Thermoacidophilic Crenarchaeon .

作者信息

Lee Junho, Lee Areum, Moon Keumok, Choi Kyoung-Hwa, Cha Jaeho

机构信息

Department of Microbiology, College of Natural Sciences, Pusan National University, Busan 46241, Republic of Korea.

Microbiological Resource Research Institute, Pusan National University, Busan 46241, Republic of Korea.

出版信息

J Microbiol Biotechnol. 2018 Jun 28;28(6):909-916. doi: 10.4014/jmb.1802.02038.

Abstract

Previously, a cytosolic trehalase (TreH) from the hyperthermophilic archaeon was reported; however, the gene responsible for the trehalase activity was not identified. Two genes, and , that encode the glycoside hydrolase family 15 type glucoamylase-like proteins in were targeted and expressed in , and their abilities to hydrolyze trehalose were examined. Recombinant Saci_1816 hydrolyzed trehalose exclusively without any help from a cofactor. The mass spectrometric analysis of partially purified native TreH also confirmed that Saci_1816 was involved in proteins exhibiting trehalase activity. Optimal trehalose hydrolysis activity of the recombinant Saci_1816 was observed at pH 4.0 and 60°C. The pH dependence of the recombinant enzyme was similar to that of the native enzyme, but its optimal temperature was 20-25°C lower, and its thermostability was also slightly reduced. From the biochemical and structural results, Saci_1816 was identified as a trehalase responsible for trehalose degradation in . Identification of the gene confirms that the degradation of trehalose in species occurs via the TreH pathway.

摘要

此前,有报道称从嗜热古菌中发现了一种胞质海藻糖酶(TreH);然而,负责海藻糖酶活性的基因尚未确定。对编码糖苷水解酶家族15型葡糖淀粉酶样蛋白的两个基因进行了靶向并在中表达,并检测了它们水解海藻糖的能力。重组Saci_1816在没有任何辅助因子的情况下专门水解海藻糖。对部分纯化的天然TreH进行质谱分析也证实,Saci_1816参与了具有海藻糖酶活性的蛋白质。重组Saci_1816在pH 4.0和60°C时观察到最佳海藻糖水解活性。重组酶的pH依赖性与天然酶相似,但其最佳温度低20 - 25°C,热稳定性也略有降低。根据生化和结构结果,Saci_1816被鉴定为负责中海藻糖降解的海藻糖酶。基因的鉴定证实了物种中海藻糖的降解是通过TreH途径发生的。

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