Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in <i>Pichia pastoris</i>
Alginate is one of the most abundant polysaccharides in algae. Alginate lyase degrades alginate through a β-elimination mechanism to produce alginate oligosaccharides with special bioactivities. Improving enzyme activity and thermal stability can promote the application of alginate lyase in the indu...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-06-01
|
| Series: | Marine Drugs |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1660-3397/18/6/305 |
| _version_ | 1827715257194250240 |
|---|---|
| author | Suxiao Yang Zhemin Liu Xiaodan Fu Changliang Zhu Qing Kong Min Yang Haijin Mou |
| author_facet | Suxiao Yang Zhemin Liu Xiaodan Fu Changliang Zhu Qing Kong Min Yang Haijin Mou |
| author_sort | Suxiao Yang |
| collection | DOAJ |
| description | Alginate is one of the most abundant polysaccharides in algae. Alginate lyase degrades alginate through a β-elimination mechanism to produce alginate oligosaccharides with special bioactivities. Improving enzyme activity and thermal stability can promote the application of alginate lyase in the industrial preparation of alginate oligosaccharides. In this study, the recombinant alginate lyase cAlyM and its thermostable mutant 102C300C were expressed and characterized in <i>Pichia pastoris</i>. The specific activities of cAlyM and 102C300C were 277.1 U/mg and 249.6 U/mg, respectively. Both enzymes showed maximal activity at 50 °C and pH 8.0 and polyG preference. The half-life values of 102C300C at 45 °C and 50 °C were 2.6 times and 11.7 times the values of cAlyM, respectively. The degradation products of 102C300C with a lower degree of polymerization contained more guluronate. The oligosaccharides with a polymerization degree of 2–4 were the final hydrolytic products. Therefore, 102C300C is potentially valuable in the production of alginate oligosaccharides with specific M/G ratio and molecular weights. |
| first_indexed | 2024-03-10T19:14:49Z |
| format | Article |
| id | doaj.art-3e0de1dd6af6450490aa8cdcc6380ecb |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-03-10T19:14:49Z |
| publishDate | 2020-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-3e0de1dd6af6450490aa8cdcc6380ecb2023-11-20T03:29:10ZengMDPI AGMarine Drugs1660-33972020-06-0118630510.3390/md18060305Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in <i>Pichia pastoris</i>Suxiao Yang0Zhemin Liu1Xiaodan Fu2Changliang Zhu3Qing Kong4Min Yang5Haijin Mou6College 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, ChinaYellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Laboratory for Marine Drugs and Bioproducts of Pilot National laboratory for Marine Science and Technology, Qingdao 266071, ChinaCollege of Food Science and Engineering, Ocean University of China, Qingdao 266003, ChinaAlginate is one of the most abundant polysaccharides in algae. Alginate lyase degrades alginate through a β-elimination mechanism to produce alginate oligosaccharides with special bioactivities. Improving enzyme activity and thermal stability can promote the application of alginate lyase in the industrial preparation of alginate oligosaccharides. In this study, the recombinant alginate lyase cAlyM and its thermostable mutant 102C300C were expressed and characterized in <i>Pichia pastoris</i>. The specific activities of cAlyM and 102C300C were 277.1 U/mg and 249.6 U/mg, respectively. Both enzymes showed maximal activity at 50 °C and pH 8.0 and polyG preference. The half-life values of 102C300C at 45 °C and 50 °C were 2.6 times and 11.7 times the values of cAlyM, respectively. The degradation products of 102C300C with a lower degree of polymerization contained more guluronate. The oligosaccharides with a polymerization degree of 2–4 were the final hydrolytic products. Therefore, 102C300C is potentially valuable in the production of alginate oligosaccharides with specific M/G ratio and molecular weights.https://www.mdpi.com/1660-3397/18/6/305<i>Pichia pastoris</i>alginate lyasethermostable mutantalginate oligosaccharides |
| spellingShingle | Suxiao Yang Zhemin Liu Xiaodan Fu Changliang Zhu Qing Kong Min Yang Haijin Mou Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in <i>Pichia pastoris</i> Marine Drugs <i>Pichia pastoris</i> alginate lyase thermostable mutant alginate oligosaccharides |
| title | Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in <i>Pichia pastoris</i> |
| title_full | Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in <i>Pichia pastoris</i> |
| title_fullStr | Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in <i>Pichia pastoris</i> |
| title_full_unstemmed | Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in <i>Pichia pastoris</i> |
| title_short | Expression and Characterization of an Alginate Lyase and Its Thermostable Mutant in <i>Pichia pastoris</i> |
| title_sort | expression and characterization of an alginate lyase and its thermostable mutant in i pichia pastoris i |
| topic | <i>Pichia pastoris</i> alginate lyase thermostable mutant alginate oligosaccharides |
| url | https://www.mdpi.com/1660-3397/18/6/305 |
| work_keys_str_mv | AT suxiaoyang expressionandcharacterizationofanalginatelyaseanditsthermostablemutantinipichiapastorisi AT zheminliu expressionandcharacterizationofanalginatelyaseanditsthermostablemutantinipichiapastorisi AT xiaodanfu expressionandcharacterizationofanalginatelyaseanditsthermostablemutantinipichiapastorisi AT changliangzhu expressionandcharacterizationofanalginatelyaseanditsthermostablemutantinipichiapastorisi AT qingkong expressionandcharacterizationofanalginatelyaseanditsthermostablemutantinipichiapastorisi AT minyang expressionandcharacterizationofanalginatelyaseanditsthermostablemutantinipichiapastorisi AT haijinmou expressionandcharacterizationofanalginatelyaseanditsthermostablemutantinipichiapastorisi |