Enhanced thermostability of nattokinase by rational design of disulfide bond
Abstract Nattokinase, the thrombolytically active substance in the health food natto, nevertheless, its lower thermostability restricts its use in food and pharmaceutical applications. In this study, two heat-resistant variants of nattokinase, designated 50–109 (M1) and 15–271 (M2), were successfull...
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| Format: | Article |
| Language: | English |
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BMC
2025-03-01
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| Series: | Microbial Cell Factories |
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| Online Access: | https://doi.org/10.1186/s12934-025-02681-5 |
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| author | Kongfang Yu Liangqi Chen Yaolei Tang Aixia Ma Wenhui Zhu Hong Wang Xiyu Tang Yuan Li Jinyao Li |
| author_facet | Kongfang Yu Liangqi Chen Yaolei Tang Aixia Ma Wenhui Zhu Hong Wang Xiyu Tang Yuan Li Jinyao Li |
| author_sort | Kongfang Yu |
| collection | DOAJ |
| description | Abstract Nattokinase, the thrombolytically active substance in the health food natto, nevertheless, its lower thermostability restricts its use in food and pharmaceutical applications. In this study, two heat-resistant variants of nattokinase, designated 50–109 (M1) and 15–271 (M2), were successfully obtained by introducing a disulfide bonding strategy. Their half-lives at 55℃ were found to be 2.50-fold and 5.17-fold higher, respectively, than that of the wild type. Furthermore, the specific enzyme activities of the variants, M1 and M2, were also increased by 2.37 and 1.66-fold, respectively. Meanwhile, the combination of two mutants increased the thermostability of nattokinase by 8.0-fold. Bioinformatics analyses indicated that the enhanced thermostability of the M1 and M2 variants was due to the increased rigidity and structural contraction of the overall structure. Finally, the fermentation process of mutant M1 was optimized to increase the expression of nattokinase. Study provides substantial molecular and theoretical support for the industrial production and application of nattokinase. |
| format | Article |
| id | doaj-art-eebc03834ca148b58f77491a0fae6f24 |
| institution | DOAJ |
| issn | 1475-2859 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj-art-eebc03834ca148b58f77491a0fae6f242025-08-20T02:47:07ZengBMCMicrobial Cell Factories1475-28592025-03-0124111310.1186/s12934-025-02681-5Enhanced thermostability of nattokinase by rational design of disulfide bondKongfang Yu0Liangqi Chen1Yaolei Tang2Aixia Ma3Wenhui Zhu4Hong Wang5Xiyu Tang6Yuan Li7Jinyao Li8Institute of Materia Medica, College of Pharmacy, Xinjiang UniversityInstitute of Materia Medica, College of Pharmacy, Xinjiang UniversityThe Third People’s Hospital of Xinjiang Uygur Autonomous RegionInstitute of Materia Medica, College of Pharmacy, Xinjiang UniversityInstitute of Materia Medica, College of Pharmacy, Xinjiang UniversityInstitute of Materia Medica, College of Pharmacy, Xinjiang UniversityInstitute of Materia Medica, College of Pharmacy, Xinjiang UniversityInstitute of Materia Medica, College of Pharmacy, Xinjiang UniversityInstitute of Materia Medica, College of Pharmacy, Xinjiang UniversityAbstract Nattokinase, the thrombolytically active substance in the health food natto, nevertheless, its lower thermostability restricts its use in food and pharmaceutical applications. In this study, two heat-resistant variants of nattokinase, designated 50–109 (M1) and 15–271 (M2), were successfully obtained by introducing a disulfide bonding strategy. Their half-lives at 55℃ were found to be 2.50-fold and 5.17-fold higher, respectively, than that of the wild type. Furthermore, the specific enzyme activities of the variants, M1 and M2, were also increased by 2.37 and 1.66-fold, respectively. Meanwhile, the combination of two mutants increased the thermostability of nattokinase by 8.0-fold. Bioinformatics analyses indicated that the enhanced thermostability of the M1 and M2 variants was due to the increased rigidity and structural contraction of the overall structure. Finally, the fermentation process of mutant M1 was optimized to increase the expression of nattokinase. Study provides substantial molecular and theoretical support for the industrial production and application of nattokinase.https://doi.org/10.1186/s12934-025-02681-5NattokinaseDisulfide bondEnzyme synthesisThermostabilityBioinformatics design |
| spellingShingle | Kongfang Yu Liangqi Chen Yaolei Tang Aixia Ma Wenhui Zhu Hong Wang Xiyu Tang Yuan Li Jinyao Li Enhanced thermostability of nattokinase by rational design of disulfide bond Microbial Cell Factories Nattokinase Disulfide bond Enzyme synthesis Thermostability Bioinformatics design |
| title | Enhanced thermostability of nattokinase by rational design of disulfide bond |
| title_full | Enhanced thermostability of nattokinase by rational design of disulfide bond |
| title_fullStr | Enhanced thermostability of nattokinase by rational design of disulfide bond |
| title_full_unstemmed | Enhanced thermostability of nattokinase by rational design of disulfide bond |
| title_short | Enhanced thermostability of nattokinase by rational design of disulfide bond |
| title_sort | enhanced thermostability of nattokinase by rational design of disulfide bond |
| topic | Nattokinase Disulfide bond Enzyme synthesis Thermostability Bioinformatics design |
| url | https://doi.org/10.1186/s12934-025-02681-5 |
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