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从阿根廷洛斯埃斯塔多斯岛亚南极沉积物中分离出的产蛋白酶细菌的多样性和酶特性

Diversity and enzyme properties of protease-producing bacteria isolated from sub-Antarctic sediments of Isla de Los Estados, Argentina.

作者信息

Olivera Nelda L, Sequeiros Cynthia, Nievas Marina L

机构信息

Centro Nacional Patagónico (CENPAT, CONICET), Blvd. Brown s/n, Puerto Madryn, U9120ACF, Chubut, Argentina.

出版信息

Extremophiles. 2007 May;11(3):517-26. doi: 10.1007/s00792-007-0064-3. Epub 2007 Feb 15.

Abstract

Protease-producing bacteria isolated from sub-Antarctic marine sediments of Isla de Los Estados (Argentina) were characterized, and the thermal inactivation kinetics of their extracellular proteases compared. Isolates were affiliated with the genera Pseudoalteromonas, Shewanella, Colwellia, Planococcus, and a strain to the family Flavobacteriaceae. Colwellia strains were moderate psychrophiles (optimal growth at about 15 degrees C, maximum growth temperature at around 25 degrees C). 16S rRNA phylogenetic analysis revealed that these strains and Colwellia aestuarii form a distinct lineage within the genus. The remaining isolates were psychrotolerant and grew optimally between 20 and 25 degrees C; two of them represent potentially novel species or genus (16S rRNA < 97% sequence similarity). The thermostability of the extracellular proteases produced by the isolates was analysed, and the inactivation rate constant (k (in)), the activation energy (Ea(in)) and the activation Gibbs free energy of thermal inactivation (Delta G( * ) (in)) determined. Delta G( * ) (in), calculated at 30 degrees C, varied between 97 and 124 kJ/mol. Colwellia enzyme extracts presented the highest thermosensitivity, while the most thermostable protease activity was shown by Shewanella spp. These results demonstrated that the stability to temperature of these enzymes varies considerably among the isolates, suggesting important variations in the thermal properties of the proteases that can coexist in this environment.

摘要

对从阿根廷洛斯埃斯塔多斯岛亚南极海洋沉积物中分离出的产蛋白酶细菌进行了表征,并比较了它们胞外蛋白酶的热失活动力学。分离菌株隶属于假交替单胞菌属、希瓦氏菌属、科尔韦氏菌属、动性球菌属,还有一株属于黄杆菌科。科尔韦氏菌属菌株为中度嗜冷菌(最适生长温度约为15℃,最高生长温度约为25℃)。16S rRNA系统发育分析表明,这些菌株与河口科尔韦氏菌在该属内形成一个独特的谱系。其余分离菌株具有耐冷性,在20至25℃之间生长最佳;其中两株代表潜在的新物种或新属(16S rRNA序列相似性<97%)。分析了分离菌株产生的胞外蛋白酶的热稳定性,并测定了失活速率常数(k(in))、活化能(Ea(in))和热失活的活化吉布斯自由能(ΔG*(in))。在30℃下计算的ΔG*(in)在97至124 kJ/mol之间变化。科尔韦氏菌的酶提取物表现出最高的热敏感性,而希瓦氏菌属的蛋白酶活性表现出最高的热稳定性。这些结果表明,这些酶对温度的稳定性在分离菌株之间有很大差异,这表明在这种环境中共存的蛋白酶的热性质存在重要差异。

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