Citation: Shijia Zu, Zhonglian Cao, Lunan Lv, Wanyi He, Wei Fu. Engineering high-performance uricase: Enhancing activity via channel loop mutations and tetramer stabilization[J]. Chinese Chemical Letters, ;2026, 37(11): 112195. doi: 10.1016/j.cclet.2025.112195 shu

Engineering high-performance uricase: Enhancing activity via channel loop mutations and tetramer stabilization

Shijia Zu ¹ ,
Zhonglian Cao ¹ ,
Lunan Lv ¹ ,
Wanyi He ¹ ,
Wei Fu ^*,

School of Pharmaceutical Sciences and Minhang Hospital, Fudan University, Shanghai 201203, China

wfu@fudan.edu.cn

Received Date: 31 August 2025
Revised Date: 26 November 2025
Accepted Date: 27 November 2025
Available Online: 28 November 2025

Figures(4)

Due to improvements in living standards and shifts in dietary patterns, the prevalence of hyperuricemia and gout has been steadily increasing, making it the second most common metabolic disorder after diabetes. Exogenous uricase has emerged as a promising therapeutic option for patients with severe and refractory gout, owing to its potent urate-lowering efficacy and ability to alleviate pain, thereby significantly improving patient quality of life. Pegloticase remains the only uricase-based drug approved by the U.S. Food and Drug Administration (FDA) for the treatment of chronic gout unresponsive to standard therapies. However, the current pipeline of exogenous uricase candidates is limited, and high immunogenicity continues to pose a major challenge, highlighting the urgent need for further innovation in this field. Here, we investigate the non-conserved amino acid residues located at the entrance of the uric acid channel and proposes a mutation-based strategy to enhance the catalytic activity of Arthrobacter globiformis uricase, achieving a maximum activity increase of 27%. Furthermore, the introduction of a K129C mutation enables the formation of a disulfide bond at the dimer–dimer interface within the tetramer, thereby significantly enhancing the thermal stability of the enzyme, with the tetrameric Tm increased by 5.88 ℃. The combination of activity-enhancing and thermostability-improving mutations resulted in uricase variants with both high catalytic efficiency and improved structural resilience. This study offers mechanistic insights into the structure–function relationship of the urate channel and proposes an innovative approach for engineering next-generation uricase therapeutics.
- Uricase,
- Gout,
- Rational mutant design,
- MD simulation,
- Thermostability
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