da Silva Abigail F, García-Fraga Belén, López-Seijas Jacobo, Sieiro Carmen
a Department of Functional Biology and Health Sciences , Microbiology Area, University of Vigo, Lagoas - Marcosende , Vigo , Spain.
Bioengineered. 2017 Jul 4;8(4):428-432. doi: 10.1080/21655979.2016.1249074. Epub 2016 Nov 28.
Many relevant applications have been demonstrated for chitinolytic enzymes. However, their successful exploitation depends upon the availability of strains and expression conditions that allow the production of active forms and large quantities of these enzymes. Escherichia coli has been commonly used to express and overproduce different proteins, among them chitinases. Improving the functional gene expression of chitinases is key to exploiting their potential. In a recent study, we described the effect of various parameters on the functional expression of 2 chitinases from different families, demonstrating that the effect of each of these parameters on the activity of both chitinases was specific to each enzyme. In this study, the expression of a Lactococcus lactis chitinase encoded by a new allele, ChiA1-2, was optimized. The results showed that not only the expression parameters seemed to influence protein production, solubility and activity but also the plasmid used for the expression. Herein, we describe the effect of 2 different promoters, tac and T7, on the expression of the active form of the chitinolytic enzyme.
几丁质分解酶已被证明有许多相关应用。然而，它们的成功利用取决于能否获得能够产生活性形式且大量生产这些酶的菌株和表达条件。大肠杆菌已被广泛用于表达和过量生产不同的蛋白质，其中包括几丁质酶。提高几丁质酶的功能基因表达是挖掘其潜力的关键。在最近的一项研究中，我们描述了各种参数对来自不同家族的两种几丁质酶功能表达的影响，表明这些参数对两种几丁质酶活性的影响因酶而异。在本研究中，对由新等位基因ChiA1-2编码的乳酸乳球菌几丁质酶的表达进行了优化。结果表明，不仅表达参数似乎会影响蛋白质的产生、溶解度和活性，而且用于表达的质粒也会产生影响。在此，我们描述了两种不同启动子tac和T7对几丁质分解酶活性形式表达的影响。
