Thermostable and Long-Circulating Albumin-Conjugated <i>Arthrobacter globiformis</i> Urate Oxidase
Urate oxidase derived from <i>Aspergillus flavus</i> has been investigated as a treatment for tumor lysis syndrome, hyperuricemia, and gout. However, its long-term use is limited owing to potential immunogenicity, low thermostability, and short circulation time in vivo. Recently, urate o...
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MDPI AG
2021-08-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/13/8/1298 |
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| author | Byungseop Yang Inchan Kwon |
| author_facet | Byungseop Yang Inchan Kwon |
| author_sort | Byungseop Yang |
| collection | DOAJ |
| description | Urate oxidase derived from <i>Aspergillus flavus</i> has been investigated as a treatment for tumor lysis syndrome, hyperuricemia, and gout. However, its long-term use is limited owing to potential immunogenicity, low thermostability, and short circulation time in vivo. Recently, urate oxidase isolated from <i>Arthrobacter globiformis</i> (AgUox) has been reported to be thermostable and less immunogenic than the <i>Aspergillus</i>-derived urate oxidase. Conjugation of human serum albumin (HSA) to therapeutic proteins has become a promising strategy to prolong circulation time in vivo. To develop a thermostable and long-circulating urate oxidase, we investigated the site-specific conjugation of HSA to AgUox based on site-specific incorporation of a clickable non-natural amino acid (frTet) and an inverse electron demand Diels–Alder reaction. We selected 14 sites for frTet incorporation using the ROSETTA design, a computational stability prediction program, among which AgUox containing frTet at position 196 (Ag12) exhibited enzymatic activity and thermostability comparable to those of wild-type AgUox. Furthermore, Ag12 exhibited a high HSA conjugation yield without compromising the enzymatic activity, generating well-defined HSA-conjugated AgUox (Ag12-HSA). In mice, the serum half-life of Ag12-HSA was approximately 29 h, which was roughly 17-fold longer than that of wild-type AgUox. Altogether, this novel formulated AgUox may hold enhanced therapeutic efficacy for several diseases. |
| first_indexed | 2024-03-10T08:28:45Z |
| format | Article |
| id | doaj.art-ddb0151beb1b4b88a571fa2230f3cca3 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-10T08:28:45Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-ddb0151beb1b4b88a571fa2230f3cca32023-11-22T09:15:43ZengMDPI AGPharmaceutics1999-49232021-08-01138129810.3390/pharmaceutics13081298Thermostable and Long-Circulating Albumin-Conjugated <i>Arthrobacter globiformis</i> Urate OxidaseByungseop Yang0Inchan Kwon1School of Materials Science and Engineering, Gwangju Institute of Science and Technology (GIST), Gwangju 61005, KoreaSchool of Materials Science and Engineering, Gwangju Institute of Science and Technology (GIST), Gwangju 61005, KoreaUrate oxidase derived from <i>Aspergillus flavus</i> has been investigated as a treatment for tumor lysis syndrome, hyperuricemia, and gout. However, its long-term use is limited owing to potential immunogenicity, low thermostability, and short circulation time in vivo. Recently, urate oxidase isolated from <i>Arthrobacter globiformis</i> (AgUox) has been reported to be thermostable and less immunogenic than the <i>Aspergillus</i>-derived urate oxidase. Conjugation of human serum albumin (HSA) to therapeutic proteins has become a promising strategy to prolong circulation time in vivo. To develop a thermostable and long-circulating urate oxidase, we investigated the site-specific conjugation of HSA to AgUox based on site-specific incorporation of a clickable non-natural amino acid (frTet) and an inverse electron demand Diels–Alder reaction. We selected 14 sites for frTet incorporation using the ROSETTA design, a computational stability prediction program, among which AgUox containing frTet at position 196 (Ag12) exhibited enzymatic activity and thermostability comparable to those of wild-type AgUox. Furthermore, Ag12 exhibited a high HSA conjugation yield without compromising the enzymatic activity, generating well-defined HSA-conjugated AgUox (Ag12-HSA). In mice, the serum half-life of Ag12-HSA was approximately 29 h, which was roughly 17-fold longer than that of wild-type AgUox. Altogether, this novel formulated AgUox may hold enhanced therapeutic efficacy for several diseases.https://www.mdpi.com/1999-4923/13/8/1298<i>Arthrobacter globiformis</i>gouthalf-life extensioninverse electron demand Diels-Alder reactionsite-specific albumin conjugationthermostability |
| spellingShingle | Byungseop Yang Inchan Kwon Thermostable and Long-Circulating Albumin-Conjugated <i>Arthrobacter globiformis</i> Urate Oxidase Pharmaceutics <i>Arthrobacter globiformis</i> gout half-life extension inverse electron demand Diels-Alder reaction site-specific albumin conjugation thermostability |
| title | Thermostable and Long-Circulating Albumin-Conjugated <i>Arthrobacter globiformis</i> Urate Oxidase |
| title_full | Thermostable and Long-Circulating Albumin-Conjugated <i>Arthrobacter globiformis</i> Urate Oxidase |
| title_fullStr | Thermostable and Long-Circulating Albumin-Conjugated <i>Arthrobacter globiformis</i> Urate Oxidase |
| title_full_unstemmed | Thermostable and Long-Circulating Albumin-Conjugated <i>Arthrobacter globiformis</i> Urate Oxidase |
| title_short | Thermostable and Long-Circulating Albumin-Conjugated <i>Arthrobacter globiformis</i> Urate Oxidase |
| title_sort | thermostable and long circulating albumin conjugated i arthrobacter globiformis i urate oxidase |
| topic | <i>Arthrobacter globiformis</i> gout half-life extension inverse electron demand Diels-Alder reaction site-specific albumin conjugation thermostability |
| url | https://www.mdpi.com/1999-4923/13/8/1298 |
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