Accumulation of U, Th, Pb, V, Rb, and Ag in wild Mushrooms macrolepiota procera (Scop.) Singer from Goc, Serbia
Abstract
Cellobiose dehydrogenase (CDH) from Phanerochaete chrysosporium can be used in lactobionic acid production, biosensor for lactose, biofuel cells, lignocellulose degradation, and wound-healing applications. To make it a better biocatalyst, CDH with higher activity in an immobilized form is desirable. For this purpose, CDH was expressed for the first time on the surface of S. cerevisiae EBY100 cells in an active form as a triple mutant tmCDH (D20N, A64T, V592M) and evolved further for higher activity using resazurin-based fluorescent assay. In order to decrease blank reaction of resazurin with yeast cells and to have linear correlation between enzyme activity on the cell surface and fluorescence signal, the assay was optimized with respect to resazurin concentration (0.1 mM), substrate concentration (10mMlactose and 0.08mMcellobiose), and pH (6.0). Using optimized assay an error prone PCR gene library of tmCDH was screened. Two mutants with 5 (H5) and 7 mutations (H9) were found having t...wo times higher activity than the parent tmCDH enzyme that already had improved activity compared to wild type CDH whose activity could not be detected on the surface of yeast cells.
Keywords:
Actinides / Bioindicator / Mycoremediation / Rubidium / VanadiumSource:
Environmental Science and Pollution Research, 2019, 26, 13, 13147-13158Funding / projects:
- Application of advanced oxidation processes and nanostructured oxide materials for the removal of pollutants from the environment, development and optimisation of instrumental techniques for efficiency monitoring (RS-172030)
- FoodEnTwin-Twinning of research activities for the frontier research in the fields of food, nutrition and environmental omics (EU-810752)
Note:
- Supplementary material: http://cherry.chem.bg.ac.rs/handle/123456789/3284
DOI: 10.1007/s11356-019-04723-z
ISSN: 0944-1344
WoS: 000465460000045
Scopus: 2-s2.0-85063215632
Collections
Institution/Community
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Vukojević, Vesna AU - Đurđić, Slađana Z. AU - Mutić, Jelena PY - 2019 UR - https://cherry.chem.bg.ac.rs/handle/123456789/3283 AB - Cellobiose dehydrogenase (CDH) from Phanerochaete chrysosporium can be used in lactobionic acid production, biosensor for lactose, biofuel cells, lignocellulose degradation, and wound-healing applications. To make it a better biocatalyst, CDH with higher activity in an immobilized form is desirable. For this purpose, CDH was expressed for the first time on the surface of S. cerevisiae EBY100 cells in an active form as a triple mutant tmCDH (D20N, A64T, V592M) and evolved further for higher activity using resazurin-based fluorescent assay. In order to decrease blank reaction of resazurin with yeast cells and to have linear correlation between enzyme activity on the cell surface and fluorescence signal, the assay was optimized with respect to resazurin concentration (0.1 mM), substrate concentration (10mMlactose and 0.08mMcellobiose), and pH (6.0). Using optimized assay an error prone PCR gene library of tmCDH was screened. Two mutants with 5 (H5) and 7 mutations (H9) were found having two times higher activity than the parent tmCDH enzyme that already had improved activity compared to wild type CDH whose activity could not be detected on the surface of yeast cells. T2 - Environmental Science and Pollution Research T1 - Accumulation of U, Th, Pb, V, Rb, and Ag in wild Mushrooms macrolepiota procera (Scop.) Singer from Goc, Serbia VL - 26 IS - 13 SP - 13147 EP - 13158 DO - 10.1007/s11356-019-04723-z ER -
@article{ author = "Vukojević, Vesna and Đurđić, Slađana Z. and Mutić, Jelena", year = "2019", abstract = "Cellobiose dehydrogenase (CDH) from Phanerochaete chrysosporium can be used in lactobionic acid production, biosensor for lactose, biofuel cells, lignocellulose degradation, and wound-healing applications. To make it a better biocatalyst, CDH with higher activity in an immobilized form is desirable. For this purpose, CDH was expressed for the first time on the surface of S. cerevisiae EBY100 cells in an active form as a triple mutant tmCDH (D20N, A64T, V592M) and evolved further for higher activity using resazurin-based fluorescent assay. In order to decrease blank reaction of resazurin with yeast cells and to have linear correlation between enzyme activity on the cell surface and fluorescence signal, the assay was optimized with respect to resazurin concentration (0.1 mM), substrate concentration (10mMlactose and 0.08mMcellobiose), and pH (6.0). Using optimized assay an error prone PCR gene library of tmCDH was screened. Two mutants with 5 (H5) and 7 mutations (H9) were found having two times higher activity than the parent tmCDH enzyme that already had improved activity compared to wild type CDH whose activity could not be detected on the surface of yeast cells.", journal = "Environmental Science and Pollution Research", title = "Accumulation of U, Th, Pb, V, Rb, and Ag in wild Mushrooms macrolepiota procera (Scop.) Singer from Goc, Serbia", volume = "26", number = "13", pages = "13147-13158", doi = "10.1007/s11356-019-04723-z" }
Vukojević, V., Đurđić, S. Z.,& Mutić, J.. (2019). Accumulation of U, Th, Pb, V, Rb, and Ag in wild Mushrooms macrolepiota procera (Scop.) Singer from Goc, Serbia. in Environmental Science and Pollution Research, 26(13), 13147-13158. https://doi.org/10.1007/s11356-019-04723-z
Vukojević V, Đurđić SZ, Mutić J. Accumulation of U, Th, Pb, V, Rb, and Ag in wild Mushrooms macrolepiota procera (Scop.) Singer from Goc, Serbia. in Environmental Science and Pollution Research. 2019;26(13):13147-13158. doi:10.1007/s11356-019-04723-z .
Vukojević, Vesna, Đurđić, Slađana Z., Mutić, Jelena, "Accumulation of U, Th, Pb, V, Rb, and Ag in wild Mushrooms macrolepiota procera (Scop.) Singer from Goc, Serbia" in Environmental Science and Pollution Research, 26, no. 13 (2019):13147-13158, https://doi.org/10.1007/s11356-019-04723-z . .