Generation of Synechocystis sp. PCC 6803 mutant with enhanced laccase-like activity

Cyanobacteria offer a good alternative to fungi for laccase production at industrial scales. Random mutagenesis approaches with ethyl methanesulfonate were used in combination with enzymatic assays screenings to select a mutant of Synechocystis sp. PCC 6803, A2, with enhanced extracellular laccase-l...

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Published inBioresource technology reports Vol. 20; p. 101266
Main Authors Hurtado-McCormick, Valentina, Commault, Audrey, Herdean, Andrei, Price, Shawn, Pernice, Mathieu, Ralph, Peter
Format Journal Article
LanguageEnglish
Published Elsevier Ltd 01.12.2022
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Abstract Cyanobacteria offer a good alternative to fungi for laccase production at industrial scales. Random mutagenesis approaches with ethyl methanesulfonate were used in combination with enzymatic assays screenings to select a mutant of Synechocystis sp. PCC 6803, A2, with enhanced extracellular laccase-like activity. Anthraquinone dye decolourisation assays revealed 7 % enhanced decolourisation in A2 relative to the wild type after 24 h. The comparison of microbiome structure and diversity of the wild type and A2 strains confirmed that the improved traits were due to the mutation(s) and not the associated bacteria. The newly isolated Synechocystis mutant is the first example of successful random mutagenesis of cyanobacteria for laccase production. Attempts to characterise the biochemical reaction and putative laccase in A2 strain were unsuccessful but will be the subject of further research. This study suggests that random mutagenesis as a powerful approach for generating cyanobacterial strains with enhanced laccase-like activity for prospective commercial applications. [Display omitted] •Mutant with laccase-like activity of 0.10 U mL−1 obtained by random mutagenesis.•Mutant grew 1.5 times faster than WT with 10 times higher laccase-like activity.•Improved traits of the mutant were not due to associated bacteria but the mutations.•Mutant decolourised 14 % of RBBR after 3 h, which is double than WT.•This study shows industrial applicability of cyanobacterial extracellular laccases.
AbstractList Cyanobacteria offer a good alternative to fungi for laccase production at industrial scales. Random mutagenesis approaches with ethyl methanesulfonate were used in combination with enzymatic assays screenings to select a mutant of Synechocystis sp. PCC 6803, A2, with enhanced extracellular laccase-like activity. Anthraquinone dye decolourisation assays revealed 7 % enhanced decolourisation in A2 relative to the wild type after 24 h. The comparison of microbiome structure and diversity of the wild type and A2 strains confirmed that the improved traits were due to the mutation(s) and not the associated bacteria. The newly isolated Synechocystis mutant is the first example of successful random mutagenesis of cyanobacteria for laccase production. Attempts to characterise the biochemical reaction and putative laccase in A2 strain were unsuccessful but will be the subject of further research. This study suggests that random mutagenesis as a powerful approach for generating cyanobacterial strains with enhanced laccase-like activity for prospective commercial applications. [Display omitted] •Mutant with laccase-like activity of 0.10 U mL−1 obtained by random mutagenesis.•Mutant grew 1.5 times faster than WT with 10 times higher laccase-like activity.•Improved traits of the mutant were not due to associated bacteria but the mutations.•Mutant decolourised 14 % of RBBR after 3 h, which is double than WT.•This study shows industrial applicability of cyanobacterial extracellular laccases.
Cyanobacteria offer a good alternative to fungi for laccase production at industrial scales. Random mutagenesis approaches with ethyl methanesulfonate were used in combination with enzymatic assays screenings to select a mutant of Synechocystis sp., A2, with enhanced extracellular laccase-like activity. Anthraquinone dye decolourisation assay revealed 7 % enhanced decolourisation in A2 relative to wild type after 24 h. Comparison of the microbiome composition, structure and richness of the wild type and A2 strains confirmed that the improved traits were due to the mutation(s) and not the associated bacteria. The newly isolated Synechocystis mutant is the first example of successful random mutagenesis of cyanobacteria for laccase production. Attempts to characterise the biochemical reaction and putative laccase in A2 strain were unsuccessful but will be the subject of further research. This study suggests that random mutagenesis as a powerful approach for generating cyanobacterial strains with enhanced laccase-like activity for prospective commercial applications.
ArticleNumber 101266
Author Commault, Audrey
Price, Shawn
Ralph, Peter
Pernice, Mathieu
Herdean, Andrei
Hurtado-McCormick, Valentina
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  givenname: Audrey
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  fullname: Commault, Audrey
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Keywords Dye decolourisation
Synechocystis
Mutagenesis
ABTS
Laccase
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Snippet Cyanobacteria offer a good alternative to fungi for laccase production at industrial scales. Random mutagenesis approaches with ethyl methanesulfonate were...
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SubjectTerms ABTS
anthraquinones
decolorization
Dye decolourisation
dyes
ethyl methanesulfonate
Laccase
microbiome
Mutagenesis
mutants
Synechocystis
Title Generation of Synechocystis sp. PCC 6803 mutant with enhanced laccase-like activity
URI https://dx.doi.org/10.1016/j.biteb.2022.101266
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