Effects of N-Terminal and C-Terminal Polyhistidine Tag on the Stability and Function of the Thermophilic P450 CYP119
Joint Authors
Aslantas, Yaprak
Surmeli, Nur Basak
Source
Bioinorganic Chemistry and Applications
Issue
Vol. 2019, Issue 2019 (31 Dec. 2019), pp.1-8, 8 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2019-06-20
Country of Publication
Egypt
No. of Pages
8
Main Subjects
Abstract EN
Biocatalysts are sought-after in synthesis of pharmaceuticals and agrochemicals due to their high regioselectivity and enantioselectivity.
Among biocatalysts, heme-containing cytochrome P450 (P450) oxygenases are an attractive target since they catalyze oxidation of “unactivated” carbon-hydrogen bonds with high efficiency.
CYP119 is an acidothermophilic P450 from Sulfolobus acidocaldarius, which has the potential to be widely used as a biocatalyst since it shows activity at high temperatures and low pH.
Polyhistidine tags (His-tags) are widely used to simplify purification of proteins.
However, His-tags can cause changes to protein structure and function.
Here, we demonstrate the effects of His-tags on CYP119.
To this end, the His-tags were cloned at the N-terminus or C-terminus of the CYP119, and His-tagged proteins were expressed and isolated.
The thermostability and peroxidase activity of His-tagged CYP119s were tested and compared to wild type CYP119.
Results indicated that while addition of His-tags increased the yield and simplified isolation of CYP119, they also influenced the electronic structure of active site and the activity of the protein.
We show that N-terminal His-tagged CYP119 has desirable properties and potential to be used in industrial applications, but mechanistic studies using this protein need careful interpretation since the His-tag affects electronic properties of the active site heme iron.
American Psychological Association (APA)
Aslantas, Yaprak& Surmeli, Nur Basak. 2019. Effects of N-Terminal and C-Terminal Polyhistidine Tag on the Stability and Function of the Thermophilic P450 CYP119. Bioinorganic Chemistry and Applications،Vol. 2019, no. 2019, pp.1-8.
https://search.emarefa.net/detail/BIM-1122474
Modern Language Association (MLA)
Aslantas, Yaprak& Surmeli, Nur Basak. Effects of N-Terminal and C-Terminal Polyhistidine Tag on the Stability and Function of the Thermophilic P450 CYP119. Bioinorganic Chemistry and Applications No. 2019 (2019), pp.1-8.
https://search.emarefa.net/detail/BIM-1122474
American Medical Association (AMA)
Aslantas, Yaprak& Surmeli, Nur Basak. Effects of N-Terminal and C-Terminal Polyhistidine Tag on the Stability and Function of the Thermophilic P450 CYP119. Bioinorganic Chemistry and Applications. 2019. Vol. 2019, no. 2019, pp.1-8.
https://search.emarefa.net/detail/BIM-1122474
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1122474