Characterization of a thermostable uricase derived from Thermoactinospora rubra YIM 77501T and its heat-resistant mechanism
IntroductionUricases are oxidative enzymes that catalyze the conversion of uric acid to allantoin and hydrogen peroxide, widely utilized in uric acid testing and the treatment of gout, hyperuricemia, and Tumor Lysis Syndrome (TLS). The search for uricases with long-term thermostability has become a...
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Frontiers Media S.A.
2025-06-01
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| author | Xin Yan Xin Yan Xin Yan Lan-Xin Tang Lan-Xin Tang Lan-Xin Tang Maite Ortúzar Maite Ortúzar Li-Quan Yang Li-Quan Yang Li-Quan Yang Peng Sang Peng Sang Na-Ying Jin Na-Ying Jin Lin-Hua Li Lin-Hua Li Zheng-Feng Yang Zheng-Feng Yang Zheng-Feng Yang Yan-Wen Wang Yan-Wen Wang Wen-Jun Li Wen-Jun Li Wen-Jun Li Wei Hu Wei Hu Yi-Rui Yin Yi-Rui Yin Yi-Rui Yin Yi-Rui Yin |
| author_facet | Xin Yan Xin Yan Xin Yan Lan-Xin Tang Lan-Xin Tang Lan-Xin Tang Maite Ortúzar Maite Ortúzar Li-Quan Yang Li-Quan Yang Li-Quan Yang Peng Sang Peng Sang Na-Ying Jin Na-Ying Jin Lin-Hua Li Lin-Hua Li Zheng-Feng Yang Zheng-Feng Yang Zheng-Feng Yang Yan-Wen Wang Yan-Wen Wang Wen-Jun Li Wen-Jun Li Wen-Jun Li Wei Hu Wei Hu Yi-Rui Yin Yi-Rui Yin Yi-Rui Yin Yi-Rui Yin |
| author_sort | Xin Yan |
| collection | DOAJ |
| description | IntroductionUricases are oxidative enzymes that catalyze the conversion of uric acid to allantoin and hydrogen peroxide, widely utilized in uric acid testing and the treatment of gout, hyperuricemia, and Tumor Lysis Syndrome (TLS). The search for uricases with long-term thermostability has become a significant area of research.MethodsIn this study, a uricase gene (truox) was obtained from the genome of Thermoactinospora rubra YIM 77501T, which was subsequently cloned and heterologously expressed. The resulting enzyme, TrUox, was comprehensively characterized for its biochemical properties and analyzed through molecular dynamics (MD) simulations.ResultsTrUox exhibits maximal catalytic activity at 35°C and pH 7.6 (mesophilic range). Notably, its thermostability is exceptional: the enzyme retains over 90% residual activity after 4 days of incubation at 50°C (with activity measured post-thermal treatment at 35°C) and maintains >90% activity for 10 days at physiological temperature (37°C). In vitro, 1.14 μg/mL TrUox effectively lowered serum uric acid (UA) from >700 to < 420 μM within 2 h in hyperuricemic models. MD simulations comparing TrUox with Rasburicase indicate it's more rigid/stable globally, less flexible, has fewer sub-states, and is more stable in FEL.DiscussionThese results demonstrate TrUox as a robust uricase exhibiting dual advantages of catalytic efficiency and enhanced thermostability, positioning it as a promising biocatalyst for industrial-scale production and therapeutic development. Our preliminary study into its thermostable mechanism provides a theoretical foundation for future production and research. |
| format | Article |
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| language | English |
| publishDate | 2025-06-01 |
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| spelling | doaj-art-88bc2b0301c941f3a1cf132acb4b944f2025-08-20T02:07:59ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-06-011610.3389/fmicb.2025.16158451615845Characterization of a thermostable uricase derived from Thermoactinospora rubra YIM 77501T and its heat-resistant mechanismXin Yan0Xin Yan1Xin Yan2Lan-Xin Tang3Lan-Xin Tang4Lan-Xin Tang5Maite Ortúzar6Maite Ortúzar7Li-Quan Yang8Li-Quan Yang9Li-Quan Yang10Peng Sang11Peng Sang12Na-Ying Jin13Na-Ying Jin14Lin-Hua Li15Lin-Hua Li16Zheng-Feng Yang17Zheng-Feng Yang18Zheng-Feng Yang19Yan-Wen Wang20Yan-Wen Wang21Wen-Jun Li22Wen-Jun Li23Wen-Jun Li24Wei Hu25Wei Hu26Yi-Rui Yin27Yi-Rui Yin28Yi-Rui Yin29Yi-Rui Yin30College of Agriculture and Biological Science, Dali University, Dali, ChinaXizang Key Laboratory of Plateau Fungi, Institute of Plateau Biology of Xizang Autonomous Region, Lhasa, ChinaDepartment of Cardiology, The First Affiliated Hospital of Kunming Medical University, Kunming, ChinaCollege of Agriculture and Biological Science, Dali University, Dali, ChinaXizang Key Laboratory of Plateau Fungi, Institute of Plateau Biology of Xizang Autonomous Region, Lhasa, ChinaDepartment of Cardiology, The First Affiliated Hospital of Kunming Medical University, Kunming, ChinaXizang Key Laboratory of Plateau Fungi, Institute of Plateau Biology of Xizang Autonomous Region, Lhasa, ChinaState Key Laboratory of Biocontrol, Guangdong Provincial Key Laboratory of Plant Resources and Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), School of Life Sciences, Sun Yat-sen University, Guangzhou, ChinaCollege of Agriculture and Biological Science, Dali University, Dali, ChinaKey Laboratory of Bioinformatics and Computational Biology, Department of Education of Yunnan Province, Dali University, Dali, ChinaCangshan Forest Ecosystem Observation and Research Station of Yunnan Province, Dali University, Dali, ChinaCollege of Agriculture and Biological Science, Dali University, Dali, ChinaKey Laboratory of Bioinformatics and Computational Biology, Department of Education of Yunnan Province, Dali University, Dali, ChinaCollege of Agriculture and Biological Science, Dali University, Dali, ChinaCangshan Forest Ecosystem Observation and Research Station of Yunnan Province, Dali University, Dali, ChinaCollege of Agriculture and Biological Science, Dali University, Dali, ChinaDepartment of Cardiology, The First Affiliated Hospital of Kunming Medical University, Kunming, ChinaCollege of Agriculture and Biological Science, Dali University, Dali, ChinaCangshan Forest Ecosystem Observation and Research Station of Yunnan Province, Dali University, Dali, ChinaCo-Innovation Center for Cangshan Mountain and Erhai Lake Integrated Protection and Green Development of Yunnan Province, Dali University, Dali, ChinaCollege of Agriculture and Biological Science, Dali University, Dali, ChinaKey Laboratory of Bioinformatics and Computational Biology, Department of Education of Yunnan Province, Dali University, Dali, ChinaCollege of Agriculture and Biological Science, Dali University, Dali, ChinaXizang Key Laboratory of Plateau Fungi, Institute of Plateau Biology of Xizang Autonomous Region, Lhasa, ChinaState Key Laboratory of Biocontrol, Guangdong Provincial Key Laboratory of Plant Resources and Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), School of Life Sciences, Sun Yat-sen University, Guangzhou, ChinaCollege of Agriculture and Biological Science, Dali University, Dali, ChinaDepartment of Cardiology, The First Affiliated Hospital of Kunming Medical University, Kunming, ChinaCollege of Agriculture and Biological Science, Dali University, Dali, ChinaXizang Key Laboratory of Plateau Fungi, Institute of Plateau Biology of Xizang Autonomous Region, Lhasa, ChinaState Key Laboratory of Biocontrol, Guangdong Provincial Key Laboratory of Plant Resources and Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), School of Life Sciences, Sun Yat-sen University, Guangzhou, ChinaCangshan Forest Ecosystem Observation and Research Station of Yunnan Province, Dali University, Dali, ChinaIntroductionUricases are oxidative enzymes that catalyze the conversion of uric acid to allantoin and hydrogen peroxide, widely utilized in uric acid testing and the treatment of gout, hyperuricemia, and Tumor Lysis Syndrome (TLS). The search for uricases with long-term thermostability has become a significant area of research.MethodsIn this study, a uricase gene (truox) was obtained from the genome of Thermoactinospora rubra YIM 77501T, which was subsequently cloned and heterologously expressed. The resulting enzyme, TrUox, was comprehensively characterized for its biochemical properties and analyzed through molecular dynamics (MD) simulations.ResultsTrUox exhibits maximal catalytic activity at 35°C and pH 7.6 (mesophilic range). Notably, its thermostability is exceptional: the enzyme retains over 90% residual activity after 4 days of incubation at 50°C (with activity measured post-thermal treatment at 35°C) and maintains >90% activity for 10 days at physiological temperature (37°C). In vitro, 1.14 μg/mL TrUox effectively lowered serum uric acid (UA) from >700 to < 420 μM within 2 h in hyperuricemic models. MD simulations comparing TrUox with Rasburicase indicate it's more rigid/stable globally, less flexible, has fewer sub-states, and is more stable in FEL.DiscussionThese results demonstrate TrUox as a robust uricase exhibiting dual advantages of catalytic efficiency and enhanced thermostability, positioning it as a promising biocatalyst for industrial-scale production and therapeutic development. Our preliminary study into its thermostable mechanism provides a theoretical foundation for future production and research.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1615845/fulluricaseThermoactinospora rubra YIM 77501Tthermostableheat-resistant mechanismdegrade uric acid |
| spellingShingle | Xin Yan Xin Yan Xin Yan Lan-Xin Tang Lan-Xin Tang Lan-Xin Tang Maite Ortúzar Maite Ortúzar Li-Quan Yang Li-Quan Yang Li-Quan Yang Peng Sang Peng Sang Na-Ying Jin Na-Ying Jin Lin-Hua Li Lin-Hua Li Zheng-Feng Yang Zheng-Feng Yang Zheng-Feng Yang Yan-Wen Wang Yan-Wen Wang Wen-Jun Li Wen-Jun Li Wen-Jun Li Wei Hu Wei Hu Yi-Rui Yin Yi-Rui Yin Yi-Rui Yin Yi-Rui Yin Characterization of a thermostable uricase derived from Thermoactinospora rubra YIM 77501T and its heat-resistant mechanism Frontiers in Microbiology uricase Thermoactinospora rubra YIM 77501T thermostable heat-resistant mechanism degrade uric acid |
| title | Characterization of a thermostable uricase derived from Thermoactinospora rubra YIM 77501T and its heat-resistant mechanism |
| title_full | Characterization of a thermostable uricase derived from Thermoactinospora rubra YIM 77501T and its heat-resistant mechanism |
| title_fullStr | Characterization of a thermostable uricase derived from Thermoactinospora rubra YIM 77501T and its heat-resistant mechanism |
| title_full_unstemmed | Characterization of a thermostable uricase derived from Thermoactinospora rubra YIM 77501T and its heat-resistant mechanism |
| title_short | Characterization of a thermostable uricase derived from Thermoactinospora rubra YIM 77501T and its heat-resistant mechanism |
| title_sort | characterization of a thermostable uricase derived from thermoactinospora rubra yim 77501t and its heat resistant mechanism |
| topic | uricase Thermoactinospora rubra YIM 77501T thermostable heat-resistant mechanism degrade uric acid |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1615845/full |
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