Domain-swapping of mesophilic xylanase with hyper-thermophilic glucanase
<p>Abstract</p> <p>Background</p> <p>Domain fusion is limited at enzyme one terminus. The issue was explored by swapping a mesophilic <it>Aspergillus niger</it> GH11 xylanase (Xyn) with a hyper-thermophilic <it>Thermotoga maritima</it> glucanase...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2012-06-01
|
| Series: | BMC Biotechnology |
| Subjects: | |
| Online Access: | http://www.biomedcentral.com/1472-6750/12/28 |
| _version_ | 1818146212381982720 |
|---|---|
| author | Liu Liangwei Wang Linmin Zhang Zhang Guo Xiaodan Li Xiangqian Chen Hongge |
| author_facet | Liu Liangwei Wang Linmin Zhang Zhang Guo Xiaodan Li Xiangqian Chen Hongge |
| author_sort | Liu Liangwei |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Domain fusion is limited at enzyme one terminus. The issue was explored by swapping a mesophilic <it>Aspergillus niger</it> GH11 xylanase (Xyn) with a hyper-thermophilic <it>Thermotoga maritima</it> glucanase (Glu) to construct two chimeras, Xyn-Glu and Glu-Xyn, with an intention to create thermostable xylanase containing glucanase activity.</p> <p>Results</p> <p>When expressed in <it>E. coli</it> BL21(DE3), the two chimeras exhibited bi-functional activities of xylanase and glucanase. The Xyn-Glu Xyn moiety had optimal reaction temperature (T<sub>opt</sub>) at 50 °C and thermal in-activation half-life (t<sub>1/2</sub>) at 50 °C for 47.6 min, compared to 47 °C and 17.6 min for the Xyn. The Glu-Xyn Xyn moiety had equivalent T<sub>opt</sub> to and shorter t<sub>1/2</sub> (5.2 min) than the Xyn. Both chimera Glu moieties were more thermostable than the Glu, and the three enzyme T<sub>opt</sub> values were higher than 96 °C. The Glu-Xyn Glu moiety optimal pH was 5.8, compared to 3.8 for the Xyn-Glu Glu moiety and the Glu. Both chimera two moieties cooperated with each other in degrading substrates.</p> <p>Conclusions</p> <p>Domain-swapping created different effects on each moiety properties. Fusing the Glu domain at C-terminus increased the xylanase thermostability, but fusing the Glu domain at N-terminus decreased the xylanase thermostability. Fusing the Xyn domain at either terminus increased the glucanase thermostability, and fusing the Xyn domain at C-terminus shifted the glucanase pH property 2 units higher towards alkaline environments. Fusing a domain at C-terminus contributes more to enzyme catalytic activity; whereas, fusing a bigger domain at N-terminus disturbs enzyme substrate binding affinity.</p> |
| first_indexed | 2024-12-11T12:15:46Z |
| format | Article |
| id | doaj.art-1de6f8bddb894adaa32f4642c6aa9b00 |
| institution | Directory Open Access Journal |
| issn | 1472-6750 |
| language | English |
| last_indexed | 2024-12-11T12:15:46Z |
| publishDate | 2012-06-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj.art-1de6f8bddb894adaa32f4642c6aa9b002022-12-22T01:07:40ZengBMCBMC Biotechnology1472-67502012-06-011212810.1186/1472-6750-12-28Domain-swapping of mesophilic xylanase with hyper-thermophilic glucanaseLiu LiangweiWang LinminZhang ZhangGuo XiaodanLi XiangqianChen Hongge<p>Abstract</p> <p>Background</p> <p>Domain fusion is limited at enzyme one terminus. The issue was explored by swapping a mesophilic <it>Aspergillus niger</it> GH11 xylanase (Xyn) with a hyper-thermophilic <it>Thermotoga maritima</it> glucanase (Glu) to construct two chimeras, Xyn-Glu and Glu-Xyn, with an intention to create thermostable xylanase containing glucanase activity.</p> <p>Results</p> <p>When expressed in <it>E. coli</it> BL21(DE3), the two chimeras exhibited bi-functional activities of xylanase and glucanase. The Xyn-Glu Xyn moiety had optimal reaction temperature (T<sub>opt</sub>) at 50 °C and thermal in-activation half-life (t<sub>1/2</sub>) at 50 °C for 47.6 min, compared to 47 °C and 17.6 min for the Xyn. The Glu-Xyn Xyn moiety had equivalent T<sub>opt</sub> to and shorter t<sub>1/2</sub> (5.2 min) than the Xyn. Both chimera Glu moieties were more thermostable than the Glu, and the three enzyme T<sub>opt</sub> values were higher than 96 °C. The Glu-Xyn Glu moiety optimal pH was 5.8, compared to 3.8 for the Xyn-Glu Glu moiety and the Glu. Both chimera two moieties cooperated with each other in degrading substrates.</p> <p>Conclusions</p> <p>Domain-swapping created different effects on each moiety properties. Fusing the Glu domain at C-terminus increased the xylanase thermostability, but fusing the Glu domain at N-terminus decreased the xylanase thermostability. Fusing the Xyn domain at either terminus increased the glucanase thermostability, and fusing the Xyn domain at C-terminus shifted the glucanase pH property 2 units higher towards alkaline environments. Fusing a domain at C-terminus contributes more to enzyme catalytic activity; whereas, fusing a bigger domain at N-terminus disturbs enzyme substrate binding affinity.</p>http://www.biomedcentral.com/1472-6750/12/28XylanaseGlucanaseDomain-swappingFusing |
| spellingShingle | Liu Liangwei Wang Linmin Zhang Zhang Guo Xiaodan Li Xiangqian Chen Hongge Domain-swapping of mesophilic xylanase with hyper-thermophilic glucanase BMC Biotechnology Xylanase Glucanase Domain-swapping Fusing |
| title | Domain-swapping of mesophilic xylanase with hyper-thermophilic glucanase |
| title_full | Domain-swapping of mesophilic xylanase with hyper-thermophilic glucanase |
| title_fullStr | Domain-swapping of mesophilic xylanase with hyper-thermophilic glucanase |
| title_full_unstemmed | Domain-swapping of mesophilic xylanase with hyper-thermophilic glucanase |
| title_short | Domain-swapping of mesophilic xylanase with hyper-thermophilic glucanase |
| title_sort | domain swapping of mesophilic xylanase with hyper thermophilic glucanase |
| topic | Xylanase Glucanase Domain-swapping Fusing |
| url | http://www.biomedcentral.com/1472-6750/12/28 |
| work_keys_str_mv | AT liuliangwei domainswappingofmesophilicxylanasewithhyperthermophilicglucanase AT wanglinmin domainswappingofmesophilicxylanasewithhyperthermophilicglucanase AT zhangzhang domainswappingofmesophilicxylanasewithhyperthermophilicglucanase AT guoxiaodan domainswappingofmesophilicxylanasewithhyperthermophilicglucanase AT lixiangqian domainswappingofmesophilicxylanasewithhyperthermophilicglucanase AT chenhongge domainswappingofmesophilicxylanasewithhyperthermophilicglucanase |