Enzyme Properties of a Laccase Obtained from the Transcriptome of the Marine-Derived Fungus <i>Stemphylium lucomagnoense</i>
Only a few studies have examined how marine-derived fungi and their enzymes adapt to salinity and plant biomass degradation. This work concerns the production and characterisation of an oxidative enzyme identified from the transcriptome of marine-derived fungus <i>Stemphylium lucomagnoense<...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-11-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/21/21/8402 |
| _version_ | 1797548391268352000 |
|---|---|
| author | Wissal Ben Ali Amal Ben Ayed Annick Turbé-Doan Emmanuel Bertrand Yann Mathieu Craig B. Faulds Anne Lomascolo Giuliano Sciara Eric Record Tahar Mechichi |
| author_facet | Wissal Ben Ali Amal Ben Ayed Annick Turbé-Doan Emmanuel Bertrand Yann Mathieu Craig B. Faulds Anne Lomascolo Giuliano Sciara Eric Record Tahar Mechichi |
| author_sort | Wissal Ben Ali |
| collection | DOAJ |
| description | Only a few studies have examined how marine-derived fungi and their enzymes adapt to salinity and plant biomass degradation. This work concerns the production and characterisation of an oxidative enzyme identified from the transcriptome of marine-derived fungus <i>Stemphylium lucomagnoense</i>. The laccase-encoding gene <i>Sl</i>Lac2 from <i>S. lucomagnoense</i> was cloned for heterologous expression in <i>Aspergillus niger</i> D15#26 for protein production in the extracellular medium of around 30 mg L<sup>−1</sup>. The extracellular recombinant enzyme <i>Sl</i>Lac2 was successfully produced and purified in three steps protocol: ultrafiltration, anion-exchange chromatography, and size exclusion chromatography, with a final recovery yield of 24%. <i>Sl</i>Lac2 was characterised by physicochemical properties, kinetic parameters, and ability to oxidise diverse phenolic substrates. We also studied its activity in the presence and absence of sea salt. The molecular mass of <i>Sl</i>Lac2 was about 75 kDa, consistent with that of most ascomycete fungal laccases. With syringaldazine as substrate, <i>Sl</i>Lac2 showed an optimal activity at pH 6 and retained nearly 100% of its activity when incubated at 50°C for 180 min. <i>Sl</i>Lac2 exhibited more than 50% of its activity with 5% wt/vol of sea salt. |
| first_indexed | 2024-03-10T14:59:42Z |
| format | Article |
| id | doaj.art-47ace053995e40438f615653474876a0 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T14:59:42Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-47ace053995e40438f615653474876a02023-11-20T20:18:27ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-11-012121840210.3390/ijms21218402Enzyme Properties of a Laccase Obtained from the Transcriptome of the Marine-Derived Fungus <i>Stemphylium lucomagnoense</i>Wissal Ben Ali0Amal Ben Ayed1Annick Turbé-Doan2Emmanuel Bertrand3Yann Mathieu4Craig B. Faulds5Anne Lomascolo6Giuliano Sciara7Eric Record8Tahar Mechichi9Biodiversité et Biotechnologie Fongiques, Aix-Marseille Université, INRAE, UMR1163 Marseille, FranceBiodiversité et Biotechnologie Fongiques, Aix-Marseille Université, INRAE, UMR1163 Marseille, FranceBiodiversité et Biotechnologie Fongiques, Aix-Marseille Université, INRAE, UMR1163 Marseille, FranceBiodiversité et Biotechnologie Fongiques, Aix-Marseille Université, INRAE, UMR1163 Marseille, FranceMichael Smith Laboratories, University of British Columbia, Vancouver, BC V6T 1Z4, CanadaBiodiversité et Biotechnologie Fongiques, Aix-Marseille Université, INRAE, UMR1163 Marseille, FranceBiodiversité et Biotechnologie Fongiques, Aix-Marseille Université, INRAE, UMR1163 Marseille, FranceBiodiversité et Biotechnologie Fongiques, Aix-Marseille Université, INRAE, UMR1163 Marseille, FranceBiodiversité et Biotechnologie Fongiques, Aix-Marseille Université, INRAE, UMR1163 Marseille, FranceLaboratoire de Biochimie et de Génie Enzymatique des Lipases, Ecole Nationale d’Ingénieurs de Sfax, Université de Sfax, Sfax 3029, TunisiaOnly a few studies have examined how marine-derived fungi and their enzymes adapt to salinity and plant biomass degradation. This work concerns the production and characterisation of an oxidative enzyme identified from the transcriptome of marine-derived fungus <i>Stemphylium lucomagnoense</i>. The laccase-encoding gene <i>Sl</i>Lac2 from <i>S. lucomagnoense</i> was cloned for heterologous expression in <i>Aspergillus niger</i> D15#26 for protein production in the extracellular medium of around 30 mg L<sup>−1</sup>. The extracellular recombinant enzyme <i>Sl</i>Lac2 was successfully produced and purified in three steps protocol: ultrafiltration, anion-exchange chromatography, and size exclusion chromatography, with a final recovery yield of 24%. <i>Sl</i>Lac2 was characterised by physicochemical properties, kinetic parameters, and ability to oxidise diverse phenolic substrates. We also studied its activity in the presence and absence of sea salt. The molecular mass of <i>Sl</i>Lac2 was about 75 kDa, consistent with that of most ascomycete fungal laccases. With syringaldazine as substrate, <i>Sl</i>Lac2 showed an optimal activity at pH 6 and retained nearly 100% of its activity when incubated at 50°C for 180 min. <i>Sl</i>Lac2 exhibited more than 50% of its activity with 5% wt/vol of sea salt.https://www.mdpi.com/1422-0067/21/21/8402laccase<i>Stemphylium</i>heterologous expressionenzyme propertiesalkalinesalt tolerance |
| spellingShingle | Wissal Ben Ali Amal Ben Ayed Annick Turbé-Doan Emmanuel Bertrand Yann Mathieu Craig B. Faulds Anne Lomascolo Giuliano Sciara Eric Record Tahar Mechichi Enzyme Properties of a Laccase Obtained from the Transcriptome of the Marine-Derived Fungus <i>Stemphylium lucomagnoense</i> International Journal of Molecular Sciences laccase <i>Stemphylium</i> heterologous expression enzyme properties alkaline salt tolerance |
| title | Enzyme Properties of a Laccase Obtained from the Transcriptome of the Marine-Derived Fungus <i>Stemphylium lucomagnoense</i> |
| title_full | Enzyme Properties of a Laccase Obtained from the Transcriptome of the Marine-Derived Fungus <i>Stemphylium lucomagnoense</i> |
| title_fullStr | Enzyme Properties of a Laccase Obtained from the Transcriptome of the Marine-Derived Fungus <i>Stemphylium lucomagnoense</i> |
| title_full_unstemmed | Enzyme Properties of a Laccase Obtained from the Transcriptome of the Marine-Derived Fungus <i>Stemphylium lucomagnoense</i> |
| title_short | Enzyme Properties of a Laccase Obtained from the Transcriptome of the Marine-Derived Fungus <i>Stemphylium lucomagnoense</i> |
| title_sort | enzyme properties of a laccase obtained from the transcriptome of the marine derived fungus i stemphylium lucomagnoense i |
| topic | laccase <i>Stemphylium</i> heterologous expression enzyme properties alkaline salt tolerance |
| url | https://www.mdpi.com/1422-0067/21/21/8402 |
| work_keys_str_mv | AT wissalbenali enzymepropertiesofalaccaseobtainedfromthetranscriptomeofthemarinederivedfungusistemphyliumlucomagnoensei AT amalbenayed enzymepropertiesofalaccaseobtainedfromthetranscriptomeofthemarinederivedfungusistemphyliumlucomagnoensei AT annickturbedoan enzymepropertiesofalaccaseobtainedfromthetranscriptomeofthemarinederivedfungusistemphyliumlucomagnoensei AT emmanuelbertrand enzymepropertiesofalaccaseobtainedfromthetranscriptomeofthemarinederivedfungusistemphyliumlucomagnoensei AT yannmathieu enzymepropertiesofalaccaseobtainedfromthetranscriptomeofthemarinederivedfungusistemphyliumlucomagnoensei AT craigbfaulds enzymepropertiesofalaccaseobtainedfromthetranscriptomeofthemarinederivedfungusistemphyliumlucomagnoensei AT annelomascolo enzymepropertiesofalaccaseobtainedfromthetranscriptomeofthemarinederivedfungusistemphyliumlucomagnoensei AT giulianosciara enzymepropertiesofalaccaseobtainedfromthetranscriptomeofthemarinederivedfungusistemphyliumlucomagnoensei AT ericrecord enzymepropertiesofalaccaseobtainedfromthetranscriptomeofthemarinederivedfungusistemphyliumlucomagnoensei AT taharmechichi enzymepropertiesofalaccaseobtainedfromthetranscriptomeofthemarinederivedfungusistemphyliumlucomagnoensei |