Rational Design of Alginate Lyase from <i>Microbulbifer</i> sp. Q7 to Improve Thermal Stability
Alginate lyase degrades alginate by the β-elimination mechanism to produce oligosaccharides with special bioactivities. The low thermal stability of alginate lyase limits its industrial application. In this study, introducing the disulfide bonds while using the rational design methodology e...
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MDPI AG
2019-06-01
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Series: | Marine Drugs |
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Online Access: | https://www.mdpi.com/1660-3397/17/6/378 |
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author | Min Yang Su-Xiao Yang Zhe-Min Liu Nan-Nan Li Li Li Hai-Jin Mou |
author_facet | Min Yang Su-Xiao Yang Zhe-Min Liu Nan-Nan Li Li Li Hai-Jin Mou |
author_sort | Min Yang |
collection | DOAJ |
description | Alginate lyase degrades alginate by the β-elimination mechanism to produce oligosaccharides with special bioactivities. The low thermal stability of alginate lyase limits its industrial application. In this study, introducing the disulfide bonds while using the rational design methodology enhanced the thermal stability of alginate lyase cAlyM from <i>Microbulbifer</i> sp. Q7. Enzyme catalytic sites, secondary structure, spatial configuration, and molecular dynamic simulation were comprehensively analyzed. When compared with cAlyM, the mutants D102C-A300C and G103C-T113C showed an increase by 2.25 and 1.16 h, respectively, in half-life time at 45 °C, in addition to increases by 1.7 °C and 0.4 °C in the melting temperature, respectively. The enzyme-specific activity and <i>k<sub>cat</sub></i>/<i>K<sub>m</sub></i> values of D102C-A300C were 1.8- and 1.5-times higher than those of cAlyM, respectively. The rational design strategy that was used in this study provides a valuable method for improving the thermal stability of the alginate lyase. |
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language | English |
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spelling | doaj.art-3d1df33ca81a4fc4b37d959540525cd32022-12-22T03:18:39ZengMDPI AGMarine Drugs1660-33972019-06-0117637810.3390/md17060378md17060378Rational Design of Alginate Lyase from <i>Microbulbifer</i> sp. Q7 to Improve Thermal StabilityMin Yang0Su-Xiao Yang1Zhe-Min Liu2Nan-Nan Li3Li Li4Hai-Jin Mou5College of Food Science and Engineering, Ocean University of China, Qingdao 266003, ChinaCollege of Food Science and Engineering, Ocean University of China, Qingdao 266003, ChinaCollege of Food Science and Engineering, Ocean University of China, Qingdao 266003, ChinaCollege of Food Science and Engineering, Ocean University of China, Qingdao 266003, ChinaCollege of Food Science and Engineering, Ocean University of China, Qingdao 266003, ChinaCollege of Food Science and Engineering, Ocean University of China, Qingdao 266003, ChinaAlginate lyase degrades alginate by the β-elimination mechanism to produce oligosaccharides with special bioactivities. The low thermal stability of alginate lyase limits its industrial application. In this study, introducing the disulfide bonds while using the rational design methodology enhanced the thermal stability of alginate lyase cAlyM from <i>Microbulbifer</i> sp. Q7. Enzyme catalytic sites, secondary structure, spatial configuration, and molecular dynamic simulation were comprehensively analyzed. When compared with cAlyM, the mutants D102C-A300C and G103C-T113C showed an increase by 2.25 and 1.16 h, respectively, in half-life time at 45 °C, in addition to increases by 1.7 °C and 0.4 °C in the melting temperature, respectively. The enzyme-specific activity and <i>k<sub>cat</sub></i>/<i>K<sub>m</sub></i> values of D102C-A300C were 1.8- and 1.5-times higher than those of cAlyM, respectively. The rational design strategy that was used in this study provides a valuable method for improving the thermal stability of the alginate lyase.https://www.mdpi.com/1660-3397/17/6/378alginate lyasethermal stabilityrational designdisulfide bondmolecular dynamic simulation |
spellingShingle | Min Yang Su-Xiao Yang Zhe-Min Liu Nan-Nan Li Li Li Hai-Jin Mou Rational Design of Alginate Lyase from <i>Microbulbifer</i> sp. Q7 to Improve Thermal Stability Marine Drugs alginate lyase thermal stability rational design disulfide bond molecular dynamic simulation |
title | Rational Design of Alginate Lyase from <i>Microbulbifer</i> sp. Q7 to Improve Thermal Stability |
title_full | Rational Design of Alginate Lyase from <i>Microbulbifer</i> sp. Q7 to Improve Thermal Stability |
title_fullStr | Rational Design of Alginate Lyase from <i>Microbulbifer</i> sp. Q7 to Improve Thermal Stability |
title_full_unstemmed | Rational Design of Alginate Lyase from <i>Microbulbifer</i> sp. Q7 to Improve Thermal Stability |
title_short | Rational Design of Alginate Lyase from <i>Microbulbifer</i> sp. Q7 to Improve Thermal Stability |
title_sort | rational design of alginate lyase from i microbulbifer i sp q7 to improve thermal stability |
topic | alginate lyase thermal stability rational design disulfide bond molecular dynamic simulation |
url | https://www.mdpi.com/1660-3397/17/6/378 |
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