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Cloning and Characterization of a Novel N-Acetyl-D-galactosamine-4-O-sulfate Sulfatase, SulA1, from a Marine Arthrobacter Strain

Daugbjerg Christensen, Monica ; Allahgholi, Leila LU ; Linares-Pastén, Javier A. LU orcid ; Friðjónsson, Ólafur ; Guðmundsson, Hörður ; Kale, Varsha ; Sardari, Roya R. R. LU ; Hreggviðsson, Guðmundur Ó. and Nordberg Karlsson, Eva LU orcid (2024) In Marine Drugs 22(3).
Abstract
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 sulA1 was characterized. The sulA1-gene is located upstream of a chondroitin lyase encoding gene in the genome of the marine Arthrobacter strain (MAT3885). The sulfatase was produced in Escherichia coli. 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... (More)
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 sulA1 was characterized. The sulA1-gene is located upstream of a chondroitin lyase encoding gene in the genome of the marine Arthrobacter strain (MAT3885). The sulfatase was produced in Escherichia coli. 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 Ca2+, and conserved residues for Ca2+ binding were identified (Asp17, Asp18, Asp277, and Asn278) in a structural model of the enzyme. The temperature and pH activity profiles (screened using p-nitrocatechol sulfate) were narrow, with an activity optimum at 40–50 °C and a pH optimum at pH 5.5. The Tm 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. (Less)
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author
; ; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Marine Drugs
volume
22
issue
3
publisher
MDPI AG
external identifiers
  • scopus:85188891461
ISSN
1660-3397
DOI
10.3390/md22030104
language
English
LU publication?
yes
id
db875860-8820-46af-b170-29f97c10c278
date added to LUP
2024-02-25 18:48:51
date last changed
2024-04-16 14:24:59
@article{db875860-8820-46af-b170-29f97c10c278,
  abstract     = {{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 sulA1 was characterized. The sulA1-gene is located upstream of a chondroitin lyase encoding gene in the genome of the marine Arthrobacter strain (MAT3885). The sulfatase was produced in Escherichia coli. 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 p-nitrocatechol sulfate) were narrow, with an activity optimum at 40–50 °C and a pH optimum at pH 5.5. The T<sub>m</sub> 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.}},
  author       = {{Daugbjerg Christensen, Monica and Allahgholi, Leila and Linares-Pastén, Javier A. and Friðjónsson, Ólafur and Guðmundsson, Hörður and Kale, Varsha and Sardari, Roya R. R. and Hreggviðsson, Guðmundur Ó. and Nordberg Karlsson, Eva}},
  issn         = {{1660-3397}},
  language     = {{eng}},
  month        = {{02}},
  number       = {{3}},
  publisher    = {{MDPI AG}},
  series       = {{Marine Drugs}},
  title        = {{Cloning and Characterization of a Novel <i>N</i>-Acetyl-D-galactosamine-4-<i>O</i>-sulfate Sulfatase, SulA1, from a Marine <i>Arthrobacter </i>Strain}},
  url          = {{http://dx.doi.org/10.3390/md22030104}},
  doi          = {{10.3390/md22030104}},
  volume       = {{22}},
  year         = {{2024}},
}