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Microcystin-Detoxifying Recombinant Saccharomyces cerevisiae Expressing the mlrA Gene from Sphingosinicella microcystinivorans B9

Contamination of water by microcystins is a global problem. These potent hepatotoxins demand constant monitoring and control methods in potable water. Promising approaches to reduce contamination risks have focused on natural microcystin biodegradation led by enzymes encoded by the mlrABCD genes. Th...

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Autores principales: Silva, Fernando de Godoi, Lopes, Daiane Dias, Hector, Ronald E., do Nascimento, Maikon Thiago, Miguel, Tatiana de Ávila, Kuroda, Emília Kiyomi, de Nóbrega, Gisele Maria de Andrade, Harada, Ken-Ichi, Hirooka, Elisa Yoko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10058252/
https://www.ncbi.nlm.nih.gov/pubmed/36985150
http://dx.doi.org/10.3390/microorganisms11030575
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author Silva, Fernando de Godoi
Lopes, Daiane Dias
Hector, Ronald E.
do Nascimento, Maikon Thiago
Miguel, Tatiana de Ávila
Kuroda, Emília Kiyomi
de Nóbrega, Gisele Maria de Andrade
Harada, Ken-Ichi
Hirooka, Elisa Yoko
author_facet Silva, Fernando de Godoi
Lopes, Daiane Dias
Hector, Ronald E.
do Nascimento, Maikon Thiago
Miguel, Tatiana de Ávila
Kuroda, Emília Kiyomi
de Nóbrega, Gisele Maria de Andrade
Harada, Ken-Ichi
Hirooka, Elisa Yoko
author_sort Silva, Fernando de Godoi
collection PubMed
description Contamination of water by microcystins is a global problem. These potent hepatotoxins demand constant monitoring and control methods in potable water. Promising approaches to reduce contamination risks have focused on natural microcystin biodegradation led by enzymes encoded by the mlrABCD genes. The first enzyme of this system (mlrA) linearizes microcystin structure, reducing toxicity and stability. Heterologous expression of mlrA in different microorganisms may enhance its production and activity, promote additional knowledge on the enzyme, and support feasible applications. In this context, we intended to express the mlrA gene from Sphingosinicella microcystinivorans B9 in an industrial Saccharomyces cerevisiae strain as an innovative biological alternative to degrade microcystins. The mlrA gene was codon-optimized for expression in yeast, and either expressed from a plasmid or through chromosomal integration at the URA3 locus. Recombinant and wild yeasts were cultivated in medium contaminated with microcystins, and the toxin content was analyzed during growth. Whereas no difference in microcystins content was observed in cultivation with the chromosomally integrated strain, the yeast strain hosting the mlrA expression plasmid reduced 83% of toxins within 120 h of cultivation. Our results show microcystinase A expressed by industrial yeast strains as a viable option for practical applications in water treatment.
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spelling pubmed-100582522023-03-30 Microcystin-Detoxifying Recombinant Saccharomyces cerevisiae Expressing the mlrA Gene from Sphingosinicella microcystinivorans B9 Silva, Fernando de Godoi Lopes, Daiane Dias Hector, Ronald E. do Nascimento, Maikon Thiago Miguel, Tatiana de Ávila Kuroda, Emília Kiyomi de Nóbrega, Gisele Maria de Andrade Harada, Ken-Ichi Hirooka, Elisa Yoko Microorganisms Article Contamination of water by microcystins is a global problem. These potent hepatotoxins demand constant monitoring and control methods in potable water. Promising approaches to reduce contamination risks have focused on natural microcystin biodegradation led by enzymes encoded by the mlrABCD genes. The first enzyme of this system (mlrA) linearizes microcystin structure, reducing toxicity and stability. Heterologous expression of mlrA in different microorganisms may enhance its production and activity, promote additional knowledge on the enzyme, and support feasible applications. In this context, we intended to express the mlrA gene from Sphingosinicella microcystinivorans B9 in an industrial Saccharomyces cerevisiae strain as an innovative biological alternative to degrade microcystins. The mlrA gene was codon-optimized for expression in yeast, and either expressed from a plasmid or through chromosomal integration at the URA3 locus. Recombinant and wild yeasts were cultivated in medium contaminated with microcystins, and the toxin content was analyzed during growth. Whereas no difference in microcystins content was observed in cultivation with the chromosomally integrated strain, the yeast strain hosting the mlrA expression plasmid reduced 83% of toxins within 120 h of cultivation. Our results show microcystinase A expressed by industrial yeast strains as a viable option for practical applications in water treatment. MDPI 2023-02-24 /pmc/articles/PMC10058252/ /pubmed/36985150 http://dx.doi.org/10.3390/microorganisms11030575 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Silva, Fernando de Godoi
Lopes, Daiane Dias
Hector, Ronald E.
do Nascimento, Maikon Thiago
Miguel, Tatiana de Ávila
Kuroda, Emília Kiyomi
de Nóbrega, Gisele Maria de Andrade
Harada, Ken-Ichi
Hirooka, Elisa Yoko
Microcystin-Detoxifying Recombinant Saccharomyces cerevisiae Expressing the mlrA Gene from Sphingosinicella microcystinivorans B9
title Microcystin-Detoxifying Recombinant Saccharomyces cerevisiae Expressing the mlrA Gene from Sphingosinicella microcystinivorans B9
title_full Microcystin-Detoxifying Recombinant Saccharomyces cerevisiae Expressing the mlrA Gene from Sphingosinicella microcystinivorans B9
title_fullStr Microcystin-Detoxifying Recombinant Saccharomyces cerevisiae Expressing the mlrA Gene from Sphingosinicella microcystinivorans B9
title_full_unstemmed Microcystin-Detoxifying Recombinant Saccharomyces cerevisiae Expressing the mlrA Gene from Sphingosinicella microcystinivorans B9
title_short Microcystin-Detoxifying Recombinant Saccharomyces cerevisiae Expressing the mlrA Gene from Sphingosinicella microcystinivorans B9
title_sort microcystin-detoxifying recombinant saccharomyces cerevisiae expressing the mlra gene from sphingosinicella microcystinivorans b9
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10058252/
https://www.ncbi.nlm.nih.gov/pubmed/36985150
http://dx.doi.org/10.3390/microorganisms11030575
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