Consensus design for improved thermostability of lipoxygenase from Anabaena sp. PCC 7120
Abstract Background Lipoxygenase (LOX) from Anabaena sp. PCC 7120 (Ana-rLOX) offers important applications in the food industry, especially for improving aroma and dough rheological properties. However, industrial applications of LOXs have been limited by their poor thermostability. Herein, we repor...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-09-01
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| Series: | BMC Biotechnology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12896-018-0468-4 |
| _version_ | 1818596910533967872 |
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| author | Hui Qian Chong Zhang Zhaoxin Lu Bingjie Xia Xiaomei Bie Haizhen Zhao Fengxia Lu Guang-Yu Yang |
| author_facet | Hui Qian Chong Zhang Zhaoxin Lu Bingjie Xia Xiaomei Bie Haizhen Zhao Fengxia Lu Guang-Yu Yang |
| author_sort | Hui Qian |
| collection | DOAJ |
| description | Abstract Background Lipoxygenase (LOX) from Anabaena sp. PCC 7120 (Ana-rLOX) offers important applications in the food industry, especially for improving aroma and dough rheological properties. However, industrial applications of LOXs have been limited by their poor thermostability. Herein, we report a bioinformatics-based consensus concept approach for the engineering of thermostable Ana-rLOX. Results A series of mutations (N130D, G260A, S437T, N130D/G260Q, N130D/S437Y) showed higher thermostability and activity than the wild-type enzyme. Thus, N130D/G260Q exhibited a 6.6-fold increase in half-life and 2.45 °C increase in unfolding temperature; N130D/S437Y showed a 10 °C increase in optimal temperature. The secondary structure did not change much that contributed to improved thermostability were investigated in detail using circular dichroism. Homology modeling suggested that enhanced thermostability and specific activity may result from favorable hydrophobic interactions. Conclusions A series of mutations were achieved, showing higher thermostability and activity than the wild-type enzyme by semi-rational mutagenesis with limited structure information. Our findings provide important new insights into molecular modifications aimed at improving Ana-rLOX thermostability and activity. |
| first_indexed | 2024-12-16T11:39:25Z |
| format | Article |
| id | doaj.art-c477ce2b004f47dfb09a71ba15cd36cb |
| institution | Directory Open Access Journal |
| issn | 1472-6750 |
| language | English |
| last_indexed | 2024-12-16T11:39:25Z |
| publishDate | 2018-09-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj.art-c477ce2b004f47dfb09a71ba15cd36cb2022-12-21T22:32:59ZengBMCBMC Biotechnology1472-67502018-09-011811710.1186/s12896-018-0468-4Consensus design for improved thermostability of lipoxygenase from Anabaena sp. PCC 7120Hui Qian0Chong Zhang1Zhaoxin Lu2Bingjie Xia3Xiaomei Bie4Haizhen Zhao5Fengxia Lu6Guang-Yu Yang7College of Food Science and Technology, Nanjing Agricultural UniversityCollege of Food Science and Technology, Nanjing Agricultural UniversityCollege of Food Science and Technology, Nanjing Agricultural UniversityCollege of Food Science and Technology, Nanjing Agricultural UniversityCollege of Food Science and Technology, Nanjing Agricultural UniversityCollege of Food Science and Technology, Nanjing Agricultural UniversityCollege of Food Science and Technology, Nanjing Agricultural UniversityState Key Laboratory of Microbial Metabolism, College of Life Science and Biotechnology, Shanghai Jiao Tong UniversityAbstract Background Lipoxygenase (LOX) from Anabaena sp. PCC 7120 (Ana-rLOX) offers important applications in the food industry, especially for improving aroma and dough rheological properties. However, industrial applications of LOXs have been limited by their poor thermostability. Herein, we report a bioinformatics-based consensus concept approach for the engineering of thermostable Ana-rLOX. Results A series of mutations (N130D, G260A, S437T, N130D/G260Q, N130D/S437Y) showed higher thermostability and activity than the wild-type enzyme. Thus, N130D/G260Q exhibited a 6.6-fold increase in half-life and 2.45 °C increase in unfolding temperature; N130D/S437Y showed a 10 °C increase in optimal temperature. The secondary structure did not change much that contributed to improved thermostability were investigated in detail using circular dichroism. Homology modeling suggested that enhanced thermostability and specific activity may result from favorable hydrophobic interactions. Conclusions A series of mutations were achieved, showing higher thermostability and activity than the wild-type enzyme by semi-rational mutagenesis with limited structure information. Our findings provide important new insights into molecular modifications aimed at improving Ana-rLOX thermostability and activity.http://link.springer.com/article/10.1186/s12896-018-0468-4LipoxygenaseThermostabilitySpecific activityConsensus concept |
| spellingShingle | Hui Qian Chong Zhang Zhaoxin Lu Bingjie Xia Xiaomei Bie Haizhen Zhao Fengxia Lu Guang-Yu Yang Consensus design for improved thermostability of lipoxygenase from Anabaena sp. PCC 7120 BMC Biotechnology Lipoxygenase Thermostability Specific activity Consensus concept |
| title | Consensus design for improved thermostability of lipoxygenase from Anabaena sp. PCC 7120 |
| title_full | Consensus design for improved thermostability of lipoxygenase from Anabaena sp. PCC 7120 |
| title_fullStr | Consensus design for improved thermostability of lipoxygenase from Anabaena sp. PCC 7120 |
| title_full_unstemmed | Consensus design for improved thermostability of lipoxygenase from Anabaena sp. PCC 7120 |
| title_short | Consensus design for improved thermostability of lipoxygenase from Anabaena sp. PCC 7120 |
| title_sort | consensus design for improved thermostability of lipoxygenase from anabaena sp pcc 7120 |
| topic | Lipoxygenase Thermostability Specific activity Consensus concept |
| url | http://link.springer.com/article/10.1186/s12896-018-0468-4 |
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