Single-step purification and characterization of Pseudomonas aeruginosa azurin.

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388955%3A_____%2F24%3A00597139" target="_blank" >RIV/61388955:_____/24:00597139 - isvavai.cz</a>
Alternative codes found
RIV/00216208:11310/24:10487642 RIV/00216208:11320/24:10487642
Result on the web
<a href="https://hdl.handle.net/11104/0355432" target="_blank" >https://hdl.handle.net/11104/0355432</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.pep.2024.106566" target="_blank" >10.1016/j.pep.2024.106566</a>

Result language
angličtina
Original language name
Single-step purification and characterization of Pseudomonas aeruginosa azurin.
Original language description
Azurin is a small periplasmic blue copper protein found in bacterial strains such as Pseudomonas and Alcaligenes where it facilitates denitrification. Azurin is extensively studied for its ability to mediate electron-transfer processes, but it has also sparked interest of the pharmaceutical community as a potential antimicrobial or anticancer agent. Here we offer a novel approach for expression and single-step purification of azurin in Escherichia coli with high yields and optimal metalation. A fusion tag strategy using an N-terminal GST tag was employed to obtain pure protein without requiring any additional purification steps. After the on-column cleavage by HRV 3C Protease, azurin is collected and additionally incubated with copper sulphate to ensure sufficient metalation. UV-VIS absorption, mass spectroscopy, and circular dichroism analysis all validated the effective production of azurin, appropriate protein folding and the development of an active site with an associated cofactor. MD simulations verified that incorporation of the N-terminal GPLGS segment does not affect azurin structure. In addition, the biological activity of azurin was tested in HeLa cells.
Czech name
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Czech description
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Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10403 - Physical chemistry

Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year
2024
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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