Cloning and Characterization of a Novel <i>N</i>-Acetyl-<span style="font-variant: small-caps">D</span>-galactosamine-4-<i>O</i>-sulfate Sulfatase, SulA1, from a Marine <i>Arthrobacter</i> Strain
Sulfation is gaining increased interest due to the role of sulfate in the bioactivity of many polysaccharides of marine origin. Hence, sulfatases, enzymes that control the degree of sulfation, are being more extensively researched. In this work, a novel sulfatase (SulA1) encoded by the gene <i>...
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| Format: | Article |
| Language: | English |
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MDPI AG
2024-02-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/22/3/104 |
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| author | Monica Daugbjerg Christensen Leila Allahgholi Javier A. Linares-Pastén Ólafur Friðjónsson Hörður Guðmundsson Varsha Kale Roya R. R. Sardari Guðmundur Ó. Hreggviðsson Eva Nordberg Karlsson |
| author_facet | Monica Daugbjerg Christensen Leila Allahgholi Javier A. Linares-Pastén Ólafur Friðjónsson Hörður Guðmundsson Varsha Kale Roya R. R. Sardari Guðmundur Ó. Hreggviðsson Eva Nordberg Karlsson |
| author_sort | Monica Daugbjerg Christensen |
| collection | DOAJ |
| description | Sulfation is gaining increased interest due to the role of sulfate in the bioactivity of many polysaccharides of marine origin. Hence, sulfatases, enzymes that control the degree of sulfation, are being more extensively researched. In this work, a novel sulfatase (SulA1) encoded by the gene <i>sulA1</i> was characterized. The <i>sulA1</i>-gene is located upstream of a chondroitin lyase encoding gene in the genome of the marine Arthrobacter strain (MAT3885). The sulfatase was produced in <i>Escherichia coli</i>. Based on the primary sequence, the enzyme is classified under sulfatase family 1 and the two catalytic residues typical of the sulfatase 1 family—Cys57 (post-translationally modified to formyl glycine for function) and His190—were conserved. The enzyme showed increased activity, but not improved stability, in the presence of Ca<sup>2+</sup>, and conserved residues for Ca<sup>2+</sup> binding were identified (Asp17, Asp18, Asp277, and Asn278) in a structural model of the enzyme. The temperature and pH activity profiles (screened using <i>p</i>-nitrocatechol sulfate) were narrow, with an activity optimum at 40–50 °C and a pH optimum at pH 5.5. The <i>T<sub>m</sub></i> was significantly higher (67 °C) than the activity optimum. Desulfation activity was not detected on polymeric substrates, but was found on GalNAc4S, which is a sulfated monomer in the repeated disaccharide unit (GlcA–GalNAc4S) of, e.g., chondroitin sulfate A. The position of the sulA1 gene upstream of a chondroitin lyase gene and combined with the activity on GalNAc4S suggests that there is an involvement of the enzyme in the chondroitin-degrading cascade reaction, which specifically removes sulfate from monomeric GalNAc4S from chondroitin sulfate degradation products. |
| first_indexed | 2024-04-24T18:03:56Z |
| format | Article |
| id | doaj.art-724b7251d85a4e73b240bec8e0ad0115 |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-04-24T18:03:56Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-724b7251d85a4e73b240bec8e0ad01152024-03-27T13:52:09ZengMDPI AGMarine Drugs1660-33972024-02-0122310410.3390/md22030104Cloning and Characterization of a Novel <i>N</i>-Acetyl-<span style="font-variant: small-caps">D</span>-galactosamine-4-<i>O</i>-sulfate Sulfatase, SulA1, from a Marine <i>Arthrobacter</i> StrainMonica Daugbjerg Christensen0Leila Allahgholi1Javier A. Linares-Pastén2Ólafur Friðjónsson3Hörður Guðmundsson4Varsha Kale5Roya R. R. Sardari6Guðmundur Ó. Hreggviðsson7Eva Nordberg Karlsson8Matís, Vínlandsleið 12, 113 Reykjavik, IcelandDivision of Biotechnology, Department of Chemistry, Lund University, P.O. Box 124, 22100 Lund, SwedenDivision of Biotechnology, Department of Chemistry, Lund University, P.O. Box 124, 22100 Lund, SwedenMatís, Vínlandsleið 12, 113 Reykjavik, IcelandMatís, Vínlandsleið 12, 113 Reykjavik, IcelandMatís, Vínlandsleið 12, 113 Reykjavik, IcelandDivision of Biotechnology, Department of Chemistry, Lund University, P.O. Box 124, 22100 Lund, SwedenMatís, Vínlandsleið 12, 113 Reykjavik, IcelandDivision of Biotechnology, Department of Chemistry, Lund University, P.O. Box 124, 22100 Lund, SwedenSulfation is gaining increased interest due to the role of sulfate in the bioactivity of many polysaccharides of marine origin. Hence, sulfatases, enzymes that control the degree of sulfation, are being more extensively researched. In this work, a novel sulfatase (SulA1) encoded by the gene <i>sulA1</i> was characterized. The <i>sulA1</i>-gene is located upstream of a chondroitin lyase encoding gene in the genome of the marine Arthrobacter strain (MAT3885). The sulfatase was produced in <i>Escherichia coli</i>. Based on the primary sequence, the enzyme is classified under sulfatase family 1 and the two catalytic residues typical of the sulfatase 1 family—Cys57 (post-translationally modified to formyl glycine for function) and His190—were conserved. The enzyme showed increased activity, but not improved stability, in the presence of Ca<sup>2+</sup>, and conserved residues for Ca<sup>2+</sup> binding were identified (Asp17, Asp18, Asp277, and Asn278) in a structural model of the enzyme. The temperature and pH activity profiles (screened using <i>p</i>-nitrocatechol sulfate) were narrow, with an activity optimum at 40–50 °C and a pH optimum at pH 5.5. The <i>T<sub>m</sub></i> was significantly higher (67 °C) than the activity optimum. Desulfation activity was not detected on polymeric substrates, but was found on GalNAc4S, which is a sulfated monomer in the repeated disaccharide unit (GlcA–GalNAc4S) of, e.g., chondroitin sulfate A. The position of the sulA1 gene upstream of a chondroitin lyase gene and combined with the activity on GalNAc4S suggests that there is an involvement of the enzyme in the chondroitin-degrading cascade reaction, which specifically removes sulfate from monomeric GalNAc4S from chondroitin sulfate degradation products.https://www.mdpi.com/1660-3397/22/3/104marine bacteriumchondroitin sulfate<i>N</i>-acetylgalactosamine-4-<i>O</i>-sulfatesulfatase |
| spellingShingle | Monica Daugbjerg Christensen Leila Allahgholi Javier A. Linares-Pastén Ólafur Friðjónsson Hörður Guðmundsson Varsha Kale Roya R. R. Sardari Guðmundur Ó. Hreggviðsson Eva Nordberg Karlsson Cloning and Characterization of a Novel <i>N</i>-Acetyl-<span style="font-variant: small-caps">D</span>-galactosamine-4-<i>O</i>-sulfate Sulfatase, SulA1, from a Marine <i>Arthrobacter</i> Strain Marine Drugs marine bacterium chondroitin sulfate <i>N</i>-acetylgalactosamine-4-<i>O</i>-sulfate sulfatase |
| title | Cloning and Characterization of a Novel <i>N</i>-Acetyl-<span style="font-variant: small-caps">D</span>-galactosamine-4-<i>O</i>-sulfate Sulfatase, SulA1, from a Marine <i>Arthrobacter</i> Strain |
| title_full | Cloning and Characterization of a Novel <i>N</i>-Acetyl-<span style="font-variant: small-caps">D</span>-galactosamine-4-<i>O</i>-sulfate Sulfatase, SulA1, from a Marine <i>Arthrobacter</i> Strain |
| title_fullStr | Cloning and Characterization of a Novel <i>N</i>-Acetyl-<span style="font-variant: small-caps">D</span>-galactosamine-4-<i>O</i>-sulfate Sulfatase, SulA1, from a Marine <i>Arthrobacter</i> Strain |
| title_full_unstemmed | Cloning and Characterization of a Novel <i>N</i>-Acetyl-<span style="font-variant: small-caps">D</span>-galactosamine-4-<i>O</i>-sulfate Sulfatase, SulA1, from a Marine <i>Arthrobacter</i> Strain |
| title_short | Cloning and Characterization of a Novel <i>N</i>-Acetyl-<span style="font-variant: small-caps">D</span>-galactosamine-4-<i>O</i>-sulfate Sulfatase, SulA1, from a Marine <i>Arthrobacter</i> Strain |
| title_sort | cloning and characterization of a novel i n i acetyl span style font variant small caps d span galactosamine 4 i o i sulfate sulfatase sula1 from a marine i arthrobacter i strain |
| topic | marine bacterium chondroitin sulfate <i>N</i>-acetylgalactosamine-4-<i>O</i>-sulfate sulfatase |
| url | https://www.mdpi.com/1660-3397/22/3/104 |
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