Improvement of optimum pH and specific activity of pectate lyase from Bacillus RN.1 using loop replacement
Background: Alkaline pectate lyase plays an important role in papermaking, biological refining and wastewater treatment, but its industrial applications are largely limited owing to its low activity and poor alkali resistance.Methods: The alkaline pectate lyase BspPel from Bacillus RN.1 was heterolo...
| الحاوية / القاعدة: | Frontiers in Bioengineering and Biotechnology |
|---|---|
| المؤلفون الرئيسيون: | , , , , , , , , , , , |
| التنسيق: | مقال |
| اللغة: | الإنجليزية |
| منشور في: |
Frontiers Media S.A.
2023-07-01
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| الموضوعات: | |
| الوصول للمادة أونلاين: | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1242123/full |
| _version_ | 1851846114571976704 |
|---|---|
| author | Piwu Li Piwu Li Xiaofeng Wei Yun Wang Hui Liu Yanpeng Xu Ziyang Zhang Junlin Li Jianbin Wang Chuanzhuang Guo Songsen Sui Junqing Wang Junqing Wang Ruiming Wang Ruiming Wang |
| author_facet | Piwu Li Piwu Li Xiaofeng Wei Yun Wang Hui Liu Yanpeng Xu Ziyang Zhang Junlin Li Jianbin Wang Chuanzhuang Guo Songsen Sui Junqing Wang Junqing Wang Ruiming Wang Ruiming Wang |
| author_sort | Piwu Li |
| collection | DOAJ |
| container_title | Frontiers in Bioengineering and Biotechnology |
| description | Background: Alkaline pectate lyase plays an important role in papermaking, biological refining and wastewater treatment, but its industrial applications are largely limited owing to its low activity and poor alkali resistance.Methods: The alkaline pectate lyase BspPel from Bacillus RN.1 was heterologously expressed in Escherichia coli BL21 (DE3) and its activity and alkali resistance were improved by loop replacement. Simultaneously, the effect of R260 on enzyme alkaline tolerance was also explored.Results: Recombinant pectate lyase (BspPel-th) showed the highest activity at 60°C and pH 11.0, and showed significant stability over a wide pH range (3.0–11.0). The specific enzyme activity after purification was 139.4 U/mg, which was 4.4 times higher than that of the wild-type enzyme. BspPel-th has good affinity for apple pectin, since the Vmax and Km were 29 μmol/min. mL and 0.46 mol/L, respectively. Molecular dynamics simulation results showed that the flexibility of the loop region of BspPel-th was improved.Conclusion: The modified BspPel-th has considerable potential for industrial applications with high pH processes. |
| format | Article |
| id | doaj-art-69c1514b4d744d5b95b6b2a7b66c8e21 |
| institution | Directory of Open Access Journals |
| issn | 2296-4185 |
| language | English |
| publishDate | 2023-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| spelling | doaj-art-69c1514b4d744d5b95b6b2a7b66c8e212025-08-19T22:26:16ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852023-07-011110.3389/fbioe.2023.12421231242123Improvement of optimum pH and specific activity of pectate lyase from Bacillus RN.1 using loop replacementPiwu Li0Piwu Li1Xiaofeng Wei2Yun Wang3Hui Liu4Yanpeng Xu5Ziyang Zhang6Junlin Li7Jianbin Wang8Chuanzhuang Guo9Songsen Sui10Junqing Wang11Junqing Wang12Ruiming Wang13Ruiming Wang14State Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology, Jinan, Shandong, ChinaDepartment of Biological Engineering, Qilu University of Technology, Jinan, Shandong, ChinaDepartment of Biological Engineering, Qilu University of Technology, Jinan, Shandong, ChinaDepartment of Biological Engineering, Qilu University of Technology, Jinan, Shandong, ChinaDepartment of Biological Engineering, Qilu University of Technology, Jinan, Shandong, ChinaDepartment of Biological Engineering, Qilu University of Technology, Jinan, Shandong, ChinaDepartment of Biological Engineering, Qilu University of Technology, Jinan, Shandong, ChinaZhucheng Dongxiao Biotechnology Co. Ltd., Zhucheng, Shandong, ChinaZhucheng Dongxiao Biotechnology Co. Ltd., Zhucheng, Shandong, ChinaZhucheng Dongxiao Biotechnology Co. Ltd., Zhucheng, Shandong, ChinaZhucheng Dongxiao Biotechnology Co. Ltd., Zhucheng, Shandong, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology, Jinan, Shandong, ChinaDepartment of Biological Engineering, Qilu University of Technology, Jinan, Shandong, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP), Qilu University of Technology, Jinan, Shandong, ChinaDepartment of Biological Engineering, Qilu University of Technology, Jinan, Shandong, ChinaBackground: Alkaline pectate lyase plays an important role in papermaking, biological refining and wastewater treatment, but its industrial applications are largely limited owing to its low activity and poor alkali resistance.Methods: The alkaline pectate lyase BspPel from Bacillus RN.1 was heterologously expressed in Escherichia coli BL21 (DE3) and its activity and alkali resistance were improved by loop replacement. Simultaneously, the effect of R260 on enzyme alkaline tolerance was also explored.Results: Recombinant pectate lyase (BspPel-th) showed the highest activity at 60°C and pH 11.0, and showed significant stability over a wide pH range (3.0–11.0). The specific enzyme activity after purification was 139.4 U/mg, which was 4.4 times higher than that of the wild-type enzyme. BspPel-th has good affinity for apple pectin, since the Vmax and Km were 29 μmol/min. mL and 0.46 mol/L, respectively. Molecular dynamics simulation results showed that the flexibility of the loop region of BspPel-th was improved.Conclusion: The modified BspPel-th has considerable potential for industrial applications with high pH processes.https://www.frontiersin.org/articles/10.3389/fbioe.2023.1242123/fullpectate lyasesalkali resistanceloop replacementenzyme activitymolecular dynamics simulation |
| spellingShingle | Piwu Li Piwu Li Xiaofeng Wei Yun Wang Hui Liu Yanpeng Xu Ziyang Zhang Junlin Li Jianbin Wang Chuanzhuang Guo Songsen Sui Junqing Wang Junqing Wang Ruiming Wang Ruiming Wang Improvement of optimum pH and specific activity of pectate lyase from Bacillus RN.1 using loop replacement pectate lyases alkali resistance loop replacement enzyme activity molecular dynamics simulation |
| title | Improvement of optimum pH and specific activity of pectate lyase from Bacillus RN.1 using loop replacement |
| title_full | Improvement of optimum pH and specific activity of pectate lyase from Bacillus RN.1 using loop replacement |
| title_fullStr | Improvement of optimum pH and specific activity of pectate lyase from Bacillus RN.1 using loop replacement |
| title_full_unstemmed | Improvement of optimum pH and specific activity of pectate lyase from Bacillus RN.1 using loop replacement |
| title_short | Improvement of optimum pH and specific activity of pectate lyase from Bacillus RN.1 using loop replacement |
| title_sort | improvement of optimum ph and specific activity of pectate lyase from bacillus rn 1 using loop replacement |
| topic | pectate lyases alkali resistance loop replacement enzyme activity molecular dynamics simulation |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1242123/full |
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