<i>Ustilago maydis</i> Yeast Mutant Produces Cytosolic Melanin by Tyrosine-Tyrosinase Activity with Stain Biosorption Capability
<i>Ustilago maydis</i> is a biotrophic basidiomycete fungus that infects corn plants and works as an excellent phytopathogen model, facilitating numerous genetic transformations for studying the mechanisms of plant infection. A random mutation event in the mutant strains designed to inve...
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MDPI AG
2023-10-01
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| Online Access: | https://www.mdpi.com/2076-3417/13/20/11288 |
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| author | Cinthya Martínez-López Melissa Vázquez-Carrada Oscar Flores-Herrera Juan Pablo Pardo Dario Rafael Olicón-Hernández Guadalupe Guerra-Sánchez |
| author_facet | Cinthya Martínez-López Melissa Vázquez-Carrada Oscar Flores-Herrera Juan Pablo Pardo Dario Rafael Olicón-Hernández Guadalupe Guerra-Sánchez |
| author_sort | Cinthya Martínez-López |
| collection | DOAJ |
| description | <i>Ustilago maydis</i> is a biotrophic basidiomycete fungus that infects corn plants and works as an excellent phytopathogen model, facilitating numerous genetic transformations for studying the mechanisms of plant infection. A random mutation event in the mutant strains designed to investigate the physiological significance of two plasma membrane proton-ATPases in this model resulted in a pigmented phenotype strain. For this study, the FB2 strain and the ΔPMA1 mutant were chosen to assess the pigment, which was confirmed as melanin through thin-layer chromatography, UV, and IR spectrophotometry. The melanin was observed to accumulate in the cytosol, as evident from scanning and transmission electron microscopy, and did not interfere with normal cell growth in yeast extract peptone dextrose media or minimal media. Notably, the mutant exhibited a 25% higher melanin yield compared to wild-type cells. To analyze the melanin synthesis, the tyrosinase activity was measured in a phosphate buffer at pH 6.5. The enzyme demonstrated greater activity with tyrosine as a substrate than with L-3,4 dihydroxyphenylalanine, maintaining the same trend in ion preference. Both FB2 and ΔPMA1 mutant cells were subjected to biosorption experiments, revealing that the mutants with an excess of cytosolic melanin were capable of removing at least 50 ppm of methylene blue. In conclusion, <i>U. maydis</i> can accumulate melanin in the cytosol without adverse physiological effects and this presents biotechnological potential for dye removal. |
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| last_indexed | 2024-03-09T13:49:41Z |
| publishDate | 2023-10-01 |
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| series | Applied Sciences |
| spelling | doaj.art-047093d3a12f4728b59a6d64c45828a62023-11-30T20:51:59ZengMDPI AGApplied Sciences2076-34172023-10-0113201128810.3390/app132011288<i>Ustilago maydis</i> Yeast Mutant Produces Cytosolic Melanin by Tyrosine-Tyrosinase Activity with Stain Biosorption CapabilityCinthya Martínez-López0Melissa Vázquez-Carrada1Oscar Flores-Herrera2Juan Pablo Pardo3Dario Rafael Olicón-Hernández4Guadalupe Guerra-Sánchez5Departamento de Micriobiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prolongación de Carpio y Plan de Ayala S/N, Casco de Sto Tomás, Alcaldía Miguel Hidalgo, Mexico City CP 11340, MexicoDepartamento de Micriobiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prolongación de Carpio y Plan de Ayala S/N, Casco de Sto Tomás, Alcaldía Miguel Hidalgo, Mexico City CP 11340, MexicoDepartamento de Bioquímica, Facultad de Medicina, Universidad Autónoma de México, Circuito Interior S/N, Ciudad Universitaria, Mexico City CP 04510, MexicoDepartamento de Bioquímica, Facultad de Medicina, Universidad Autónoma de México, Circuito Interior S/N, Ciudad Universitaria, Mexico City CP 04510, MexicoDepartamento de Micriobiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prolongación de Carpio y Plan de Ayala S/N, Casco de Sto Tomás, Alcaldía Miguel Hidalgo, Mexico City CP 11340, MexicoDepartamento de Micriobiología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Prolongación de Carpio y Plan de Ayala S/N, Casco de Sto Tomás, Alcaldía Miguel Hidalgo, Mexico City CP 11340, Mexico<i>Ustilago maydis</i> is a biotrophic basidiomycete fungus that infects corn plants and works as an excellent phytopathogen model, facilitating numerous genetic transformations for studying the mechanisms of plant infection. A random mutation event in the mutant strains designed to investigate the physiological significance of two plasma membrane proton-ATPases in this model resulted in a pigmented phenotype strain. For this study, the FB2 strain and the ΔPMA1 mutant were chosen to assess the pigment, which was confirmed as melanin through thin-layer chromatography, UV, and IR spectrophotometry. The melanin was observed to accumulate in the cytosol, as evident from scanning and transmission electron microscopy, and did not interfere with normal cell growth in yeast extract peptone dextrose media or minimal media. Notably, the mutant exhibited a 25% higher melanin yield compared to wild-type cells. To analyze the melanin synthesis, the tyrosinase activity was measured in a phosphate buffer at pH 6.5. The enzyme demonstrated greater activity with tyrosine as a substrate than with L-3,4 dihydroxyphenylalanine, maintaining the same trend in ion preference. Both FB2 and ΔPMA1 mutant cells were subjected to biosorption experiments, revealing that the mutants with an excess of cytosolic melanin were capable of removing at least 50 ppm of methylene blue. In conclusion, <i>U. maydis</i> can accumulate melanin in the cytosol without adverse physiological effects and this presents biotechnological potential for dye removal.https://www.mdpi.com/2076-3417/13/20/11288melanin<i>Ustilago maydis</i>melanin accumulationtyrosinaseL-DOPA pathway |
| spellingShingle | Cinthya Martínez-López Melissa Vázquez-Carrada Oscar Flores-Herrera Juan Pablo Pardo Dario Rafael Olicón-Hernández Guadalupe Guerra-Sánchez <i>Ustilago maydis</i> Yeast Mutant Produces Cytosolic Melanin by Tyrosine-Tyrosinase Activity with Stain Biosorption Capability Applied Sciences melanin <i>Ustilago maydis</i> melanin accumulation tyrosinase L-DOPA pathway |
| title | <i>Ustilago maydis</i> Yeast Mutant Produces Cytosolic Melanin by Tyrosine-Tyrosinase Activity with Stain Biosorption Capability |
| title_full | <i>Ustilago maydis</i> Yeast Mutant Produces Cytosolic Melanin by Tyrosine-Tyrosinase Activity with Stain Biosorption Capability |
| title_fullStr | <i>Ustilago maydis</i> Yeast Mutant Produces Cytosolic Melanin by Tyrosine-Tyrosinase Activity with Stain Biosorption Capability |
| title_full_unstemmed | <i>Ustilago maydis</i> Yeast Mutant Produces Cytosolic Melanin by Tyrosine-Tyrosinase Activity with Stain Biosorption Capability |
| title_short | <i>Ustilago maydis</i> Yeast Mutant Produces Cytosolic Melanin by Tyrosine-Tyrosinase Activity with Stain Biosorption Capability |
| title_sort | i ustilago maydis i yeast mutant produces cytosolic melanin by tyrosine tyrosinase activity with stain biosorption capability |
| topic | melanin <i>Ustilago maydis</i> melanin accumulation tyrosinase L-DOPA pathway |
| url | https://www.mdpi.com/2076-3417/13/20/11288 |
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