Isolation and characterization of two serine proteases from metagenomic libraries of the Gobi and Death Valley deserts
Isolation and characterization of two serine proteases from metagenomic libraries of the Gobi and Death Valley deserts
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DOI：
10.1007/s00253-011-3256-9
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发表时间：
2011-08-01
影响因子：
5
通讯作者：
DuBow, Michael S.
中科院分区：
文献类型：
--
作者：
Neveu, Julie;Regeard, Christophe;DuBow, Michael S.
环境DNA宏基因组文库的功能活性筛选可以提供新分子和酶的重要来源。在这项研究中，我们从戈壁和死亡谷沙漠表层沙子样品的两个宏基因组文库中鉴定了17个潜在的蛋白酶产生克隆。对其中两种蛋白酶DV 1和M30进行了纯化和生化检查。这两种蛋白酶显示的分子量为41.5 kDa和45.7 kDa，分别在SDS聚丙烯酰胺凝胶。与已知蛋白酶序列的比对显示小于55%的氨基酸序列同一性。这两种丝氨酸蛋白酶似乎属于枯草杆菌蛋白酶（S8A）家族，并显示出一些独特的生化特性。蛋白酶DV 1的最适pH为8，最适活性为55 ℃，而蛋白酶M30的最适pH > 11，最适活性为40 ℃。这些酶的特性使它们对生物技术应用具有潜在的用途，并再次证明宏基因组方法可以很有用，特别是当与沙漠等新环境的研究结合起来时。
The screening of environmental DNA metagenome libraries for functional activities can provide an important source of new molecules and enzymes. In this study, we identified 17 potential protease-producing clones from two metagenomic libraries derived from samples of surface sand from the Gobi and Death Valley deserts. Two of the proteases, DV1 and M30, were purified and biochemically examined. These two proteases displayed a molecular mass of 41.5 kDa and 45.7 kDa, respectively, on SDS polyacrylamide gels. Alignments with known protease sequences showed less than 55% amino acid sequence identity. These two serine proteases appear to belong to the subtilisin (S8A) family and displayed several unique biochemical properties. Protease DV1 had an optimum pH of 8 and an optimal activity at 55A degrees C, while protease M30 had an optimum pH > 11 and optimal activity at 40A degrees C. The properties of these enzymes make them potentially useful for biotechnological applications and again demonstrate that metagenomic approaches can be useful, especially when coupled with the study of novel environments such as deserts.