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嗜热古菌火球菌K1来源的O-磷酸-L-丝氨酸巯基酶在有机溶剂中的热稳定性及反应活性

Thermostability and reactivity in organic solvent of O-phospho-L-serine sulfhydrylase from hyperthermophilic archaeon Aeropyrum pernix K1.

作者信息

Nakamura Takashi, Asai Shinji, Nakata Kaori, Kunimoto Kohei, Oguri Masateru, Ishikawa Kazuhiko

机构信息

a Faculty of Bioscience, Laboratory of Molecular Biochemistry , Nagahama Institute of Bio-Science and Technology , Nagahama , Japan.

出版信息

Biosci Biotechnol Biochem. 2015;79(8):1280-6. doi: 10.1080/09168451.2015.1020753. Epub 2015 Mar 17.

Abstract

O-phospho-l-serine sulfhydrylase (OPSS) from archaeon Aeropyrum pernix K1 is able to synthesize l-cysteine even at 80 °C. In this article, we compared thermal stability and reactivity in organic solvent of OPSS with those of O-acetyl-l-serine sulfhydrylase B (OASS-B) from Escherichia coli. As a result, the thermostability of OPSS was much higher than that of OASS-B. Moreover, the activity of OPSS increased in the reaction mixture containing the organic solvent, such as N, N'-dimethyl formamide and 1,4-dioxane, whereas that of OASS-B gradually decreased as the content of organic solvent increased. From the crystal structural analysis, the intramolecular electrostatic interactions of N-terminal domain in OPSS seemed to be correlated with the tolerance of OPSS to high temperature and organic solvent. These results indicate that OPSS is more superior to OASS-B for the industrial production of l-cysteine and unnatural amino acids that are useful pharmaceuticals in the presence of organic solvent.

摘要

来自嗜热栖热菌K1的O-磷酸-L-丝氨酸巯基酶(OPSS)即使在80°C时也能够合成L-半胱氨酸。在本文中,我们比较了OPSS与来自大肠杆菌的O-乙酰-L-丝氨酸巯基酶B(OASS-B)在有机溶剂中的热稳定性和反应活性。结果表明,OPSS的热稳定性远高于OASS-B。此外,在含有N,N'-二甲基甲酰胺和1,4-二氧六环等有机溶剂的反应混合物中,OPSS的活性增加,而OASS-B的活性则随着有机溶剂含量的增加而逐渐降低。通过晶体结构分析,OPSS中N端结构域的分子内静电相互作用似乎与OPSS对高温和有机溶剂的耐受性相关。这些结果表明,在有机溶剂存在的情况下,对于工业生产L-半胱氨酸和作为有用药物的非天然氨基酸而言,OPSS比OASS-B更具优势。

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