Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitution
Xylanase, a glycoside hydrolase, is widely used in the food, papermaking, and textile industries; however, most xylanases are inactive at high temperatures. In this study, a xylanase gene, CFXyl3, was cloned from Cellulomonas flavigena and expressed in Escherichia coli BL21 (DE3). To improve the the...
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| Format: | Article |
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Frontiers Media S.A.
2022-11-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2022.1044291/full |
| _version_ | 1811243658155917312 |
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| author | Wenzhuo Tian Wenzhuo Tian Ziyang Zhang Cuiping Yang Piwu Li Piwu Li Jing Xiao Jing Xiao Ruiming Wang Ruiming Wang Peng Du Peng Du Nan Li Nan Li Junqing Wang Junqing Wang |
| author_facet | Wenzhuo Tian Wenzhuo Tian Ziyang Zhang Cuiping Yang Piwu Li Piwu Li Jing Xiao Jing Xiao Ruiming Wang Ruiming Wang Peng Du Peng Du Nan Li Nan Li Junqing Wang Junqing Wang |
| author_sort | Wenzhuo Tian |
| collection | DOAJ |
| description | Xylanase, a glycoside hydrolase, is widely used in the food, papermaking, and textile industries; however, most xylanases are inactive at high temperatures. In this study, a xylanase gene, CFXyl3, was cloned from Cellulomonas flavigena and expressed in Escherichia coli BL21 (DE3). To improve the thermostability of xylanase, four hybrid xylanases with enhanced thermostability (designated EcsXyl1–4) were engineered from CFXyl3, guided by primary and 3D structure analyses. The optimal temperature of CFXyl3 was improved by replacing its N-terminus with the corresponding area of SyXyn11P, a xylanase that belongs to the hyperthermostable GH11 family. The optimal temperatures of the hybrid xylanases EcsXyl1–4 were 60, 60, 65, and 85°C, respectively. The optimal temperature of EcsXyl4 was 30 C higher than that of CFXyl3 (55°C) and its melting temperature was 34.5°C higher than that of CFXyl3. After the hydrolysis of beechwood xylan, the main hydrolysates were xylotetraose, xylotriose, and xylobiose; thus, these hybrid xylanases could be applied to prebiotic xylooligosaccharide manufacturing. |
| first_indexed | 2024-04-12T14:11:22Z |
| format | Article |
| id | doaj.art-c1971dd4df354a6fa481a50e22dabe2b |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-12T14:11:22Z |
| publishDate | 2022-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-c1971dd4df354a6fa481a50e22dabe2b2022-12-22T03:29:52ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-11-011010.3389/fbioe.2022.10442911044291Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitutionWenzhuo Tian0Wenzhuo Tian1Ziyang Zhang2Cuiping Yang3Piwu Li4Piwu Li5Jing Xiao6Jing Xiao7Ruiming Wang8Ruiming Wang9Peng Du10Peng Du11Nan Li12Nan Li13Junqing Wang14Junqing Wang15State Key Laboratory of Biobased Material and Green Papermaking (LBMP) (Qilu University of Technology), Jinan, ChinaSchool of Biological Engineering, Qilu University of Technology, Jinan, ChinaSchool of Biological Engineering, Qilu University of Technology, Jinan, ChinaSchool of Biological Engineering, Qilu University of Technology, Jinan, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP) (Qilu University of Technology), Jinan, ChinaSchool of Biological Engineering, Qilu University of Technology, Jinan, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP) (Qilu University of Technology), Jinan, ChinaSchool of Biological Engineering, Qilu University of Technology, Jinan, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP) (Qilu University of Technology), Jinan, ChinaSchool of Biological Engineering, Qilu University of Technology, Jinan, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP) (Qilu University of Technology), Jinan, ChinaSchool of Biological Engineering, Qilu University of Technology, Jinan, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP) (Qilu University of Technology), Jinan, ChinaSchool of Biological Engineering, Qilu University of Technology, Jinan, ChinaState Key Laboratory of Biobased Material and Green Papermaking (LBMP) (Qilu University of Technology), Jinan, ChinaSchool of Biological Engineering, Qilu University of Technology, Jinan, ChinaXylanase, a glycoside hydrolase, is widely used in the food, papermaking, and textile industries; however, most xylanases are inactive at high temperatures. In this study, a xylanase gene, CFXyl3, was cloned from Cellulomonas flavigena and expressed in Escherichia coli BL21 (DE3). To improve the thermostability of xylanase, four hybrid xylanases with enhanced thermostability (designated EcsXyl1–4) were engineered from CFXyl3, guided by primary and 3D structure analyses. The optimal temperature of CFXyl3 was improved by replacing its N-terminus with the corresponding area of SyXyn11P, a xylanase that belongs to the hyperthermostable GH11 family. The optimal temperatures of the hybrid xylanases EcsXyl1–4 were 60, 60, 65, and 85°C, respectively. The optimal temperature of EcsXyl4 was 30 C higher than that of CFXyl3 (55°C) and its melting temperature was 34.5°C higher than that of CFXyl3. After the hydrolysis of beechwood xylan, the main hydrolysates were xylotetraose, xylotriose, and xylobiose; thus, these hybrid xylanases could be applied to prebiotic xylooligosaccharide manufacturing.https://www.frontiersin.org/articles/10.3389/fbioe.2022.1044291/fullN-terminus substitutionthermostabilityxylanaseglycoside hydrolase family 11Cellulomonas flavigena |
| spellingShingle | Wenzhuo Tian Wenzhuo Tian Ziyang Zhang Cuiping Yang Piwu Li Piwu Li Jing Xiao Jing Xiao Ruiming Wang Ruiming Wang Peng Du Peng Du Nan Li Nan Li Junqing Wang Junqing Wang Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitution Frontiers in Bioengineering and Biotechnology N-terminus substitution thermostability xylanase glycoside hydrolase family 11 Cellulomonas flavigena |
| title | Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitution |
| title_full | Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitution |
| title_fullStr | Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitution |
| title_full_unstemmed | Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitution |
| title_short | Engineering mesophilic GH11 xylanase from Cellulomonas flavigena by rational design of N-terminus substitution |
| title_sort | engineering mesophilic gh11 xylanase from cellulomonas flavigena by rational design of n terminus substitution |
| topic | N-terminus substitution thermostability xylanase glycoside hydrolase family 11 Cellulomonas flavigena |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2022.1044291/full |
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