Characterization of sulfhydryl oxidase from Aspergillus tubingensis

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Nivala , O , Faccio , G , Arvas , M , Permi , P , Buchert , J , Kruus , K & Mattinen , M-L 2017 , ' Characterization of sulfhydryl oxidase from Aspergillus tubingensis ' , BMC Biochemistry , vol. 18 , 15 . https://doi.org/10.1186/s12858-017-0090-4

Title: Characterization of sulfhydryl oxidase from Aspergillus tubingensis
Author: Nivala, Outi; Faccio, Greta; Arvas, Mikko; Permi, Perttu; Buchert, Johanna; Kruus, Kristiina; Mattinen, Maija-Liisa
Other contributor: University of Helsinki, Institute of Biotechnology

Date: 2017-12-08
Language: eng
Number of pages: 14
Belongs to series: BMC Biochemistry
ISSN: 1471-2091
DOI: https://doi.org/10.1186/s12858-017-0090-4
URI: http://hdl.handle.net/10138/232820
Abstract: Background: Despite of the presence of sulfhydryl oxidases (SOXs) in the secretomes of industrially relevant organisms and their many potential applications, only few of these enzymes have been biochemically characterized. In addition, basic functions of most of the SOX enzymes reported so far are not fully understood. In particular, the physiological role of secreted fungal SOXs is unclear. Results: The recently identified SOX from Aspergillus tubingensis (AtSOX) was produced, purified and characterized in the present work. AtSOX had a pH optimum of 6.5, and showed a good pH stability retaining more than 80% of the initial activity in a pH range 4-8.5 within 20 h. More than 70% of the initial activity was retained after incubation at 50 degrees C for 20 h. AtSOX contains a non-covalently bound flavin cofactor. The enzyme oxidised a sulfhydryl group of glutathione to form a disulfide bond, as verified by nuclear magnetic resonance spectroscopy. AtSOX preferred glutathione as a substrate over cysteine and dithiothreitol. The activity of the enzyme was totally inhibited by 10 mM zinc sulphate. Peptide-and protein-bound sulfhydryl groups in bikunin, gliotoxin, holomycin, insulin B chain, and ribonuclease A, were not oxidised by the enzyme. Based on the analysis of 33 fungal genomes, SOX enzyme encoding genes were found close to nonribosomal peptide synthetases (NRPS) but not with polyketide synthases (PKS). In the phylogenetic tree, constructed from 25 SOX and thioredoxin reductase sequences from IPR000103 InterPro family, AtSOX was evolutionary closely related to other Aspergillus SOXs. Oxidoreductases involved in the maturation of nonribosomal peptides of fungal and bacterial origin, namely GliT, HlmI and DepH, were also evolutionary closely related to AtSOX whereas fungal thioreductases were more distant. Conclusions: AtSOX (55 kDa) is a fungal secreted flavin-dependent enzyme with good stability to both pH and temperature. A Michaelis-Menten behaviour was observed with reduced glutathione as a substrate. Based on the location of SOX enzyme encoding genes close to NRPSs, SOXs could be involved in the secondary metabolism and act as an accessory enzyme in the production of nonribosomal peptides.
Subject: Secreted sulfhydryl oxidase
Dithiol oxidase
Aspergillus tubingensis
Glutathione oxidation
Nonribosomal peptide synthesis
Secondary metabolism
NATURAL-PRODUCTS
DISULFIDE BONDS
BOVINE-MILK
CXXC-MOTIF
FAMILY
BIOSYNTHESIS
NIGER
IDENTIFICATION
PURIFICATION
SENSITIVITY
1182 Biochemistry, cell and molecular biology
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