Development and Characterization of Two Types of Surface Displayed Levansucrases for Levan Biosynthesis
Levan has wide applications in chemical, cosmetic, pharmaceutical and food industries. The free levansucrase is usually used in the biosynthesis of levan, but the poor reusability and low stability of free levansucrase have limited its large-scale use. To address this problem, the surface-displayed...
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MDPI AG
2021-06-01
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| Series: | Catalysts |
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| Online Access: | https://www.mdpi.com/2073-4344/11/7/757 |
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| author | Huiyi Shang Danni Yang Dairong Qiao Hui Xu Yi Cao |
| author_facet | Huiyi Shang Danni Yang Dairong Qiao Hui Xu Yi Cao |
| author_sort | Huiyi Shang |
| collection | DOAJ |
| description | Levan has wide applications in chemical, cosmetic, pharmaceutical and food industries. The free levansucrase is usually used in the biosynthesis of levan, but the poor reusability and low stability of free levansucrase have limited its large-scale use. To address this problem, the surface-displayed levansucrase in <i>Saccharomyces cerevisiae</i> were generated and evaluated in this study. The levansucrase from <i>Zymomonas mobilis</i> was displayed on the cell surface of <i>Saccharomyces cerevisiae</i> EBY100 using a various yeast surface display platform. The N-terminal fusion partner is based on a-agglutinin, and the C-terminal one is Flo1p. The yield of levan produced by these two whole-cell biocatalysts reaches 26 g/L and 34 g/L in 24 h, respectively. Meanwhile, the stability of the surface-displayed levansucrases is significantly enhanced. After six reuses, these two biocatalysts retained over 50% and 60% of their initial activities, respectively. Furthermore, the molecular weight and polydispersity test of the products suggested that the whole-cell biocatalyst of levansucrase displayed by Flo1p has more potentials in the production of levan with low molecular weight which is critical in certain applications. In conclusion, our method not only enable the possibility to reuse the enzyme, but also improves the stability of the enzyme. |
| first_indexed | 2024-03-10T10:11:11Z |
| format | Article |
| id | doaj.art-e49fc208b2b64bf8a2d3fb20a7d02272 |
| institution | Directory Open Access Journal |
| issn | 2073-4344 |
| language | English |
| last_indexed | 2024-03-10T10:11:11Z |
| publishDate | 2021-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Catalysts |
| spelling | doaj.art-e49fc208b2b64bf8a2d3fb20a7d022722023-11-22T01:11:17ZengMDPI AGCatalysts2073-43442021-06-0111775710.3390/catal11070757Development and Characterization of Two Types of Surface Displayed Levansucrases for Levan BiosynthesisHuiyi Shang0Danni Yang1Dairong Qiao2Hui Xu3Yi Cao4Microbiology and Metabolic Engineering of Key Laboratory of Sichuan Province, The College of Life Science, Sichuan University, Chengdu 610065, ChinaMicrobiology and Metabolic Engineering of Key Laboratory of Sichuan Province, The College of Life Science, Sichuan University, Chengdu 610065, ChinaMicrobiology and Metabolic Engineering of Key Laboratory of Sichuan Province, The College of Life Science, Sichuan University, Chengdu 610065, ChinaMicrobiology and Metabolic Engineering of Key Laboratory of Sichuan Province, The College of Life Science, Sichuan University, Chengdu 610065, ChinaMicrobiology and Metabolic Engineering of Key Laboratory of Sichuan Province, The College of Life Science, Sichuan University, Chengdu 610065, ChinaLevan has wide applications in chemical, cosmetic, pharmaceutical and food industries. The free levansucrase is usually used in the biosynthesis of levan, but the poor reusability and low stability of free levansucrase have limited its large-scale use. To address this problem, the surface-displayed levansucrase in <i>Saccharomyces cerevisiae</i> were generated and evaluated in this study. The levansucrase from <i>Zymomonas mobilis</i> was displayed on the cell surface of <i>Saccharomyces cerevisiae</i> EBY100 using a various yeast surface display platform. The N-terminal fusion partner is based on a-agglutinin, and the C-terminal one is Flo1p. The yield of levan produced by these two whole-cell biocatalysts reaches 26 g/L and 34 g/L in 24 h, respectively. Meanwhile, the stability of the surface-displayed levansucrases is significantly enhanced. After six reuses, these two biocatalysts retained over 50% and 60% of their initial activities, respectively. Furthermore, the molecular weight and polydispersity test of the products suggested that the whole-cell biocatalyst of levansucrase displayed by Flo1p has more potentials in the production of levan with low molecular weight which is critical in certain applications. In conclusion, our method not only enable the possibility to reuse the enzyme, but also improves the stability of the enzyme.https://www.mdpi.com/2073-4344/11/7/757levansucraseyeast surface displaywhole-cell catalyst<i>Zymomonas mobilis</i>levan |
| spellingShingle | Huiyi Shang Danni Yang Dairong Qiao Hui Xu Yi Cao Development and Characterization of Two Types of Surface Displayed Levansucrases for Levan Biosynthesis Catalysts levansucrase yeast surface display whole-cell catalyst <i>Zymomonas mobilis</i> levan |
| title | Development and Characterization of Two Types of Surface Displayed Levansucrases for Levan Biosynthesis |
| title_full | Development and Characterization of Two Types of Surface Displayed Levansucrases for Levan Biosynthesis |
| title_fullStr | Development and Characterization of Two Types of Surface Displayed Levansucrases for Levan Biosynthesis |
| title_full_unstemmed | Development and Characterization of Two Types of Surface Displayed Levansucrases for Levan Biosynthesis |
| title_short | Development and Characterization of Two Types of Surface Displayed Levansucrases for Levan Biosynthesis |
| title_sort | development and characterization of two types of surface displayed levansucrases for levan biosynthesis |
| topic | levansucrase yeast surface display whole-cell catalyst <i>Zymomonas mobilis</i> levan |
| url | https://www.mdpi.com/2073-4344/11/7/757 |
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