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Pregled bibliografske jedinice broj: 350981

Single-site oxidation, cysteine 108 to cysteine sulfinic acid, in D-amino acid oxidase from Trigonopsis variabilis and its structural and functional consequences


Slavica, Anita; Dib, Iskandar; Nidetzky, Bernd
Single-site oxidation, cysteine 108 to cysteine sulfinic acid, in D-amino acid oxidase from Trigonopsis variabilis and its structural and functional consequences // Applied and Environmental Microbiology, 71 (2005), 12; 8061-8068 (međunarodna recenzija, članak, znanstveni)


CROSBI ID: 350981 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Single-site oxidation, cysteine 108 to cysteine sulfinic acid, in D-amino acid oxidase from Trigonopsis variabilis and its structural and functional consequences

Autori
Slavica, Anita ; Dib, Iskandar ; Nidetzky, Bernd

Izvornik
Applied and Environmental Microbiology (0099-2240) 71 (2005), 12; 8061-8068

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
enzyme activity; oxidative modification; cysteine sulfinic acid; MS

Sažetak
One of the primary sources of enzyme instability is protein oxidative modification triggering activity loss or denaturation. We show here that the side chain of Cys108 is the main site undergoing stress-induced oxidation in Trigonopsis variabilis D-amino acid oxidase, a flavoenzyme employed industrially for the conversion of cephalosporin C. High-resolution anion exchange chromatography was used to separate the reduced and oxidized protein forms which constitute, in a molar ratio of about 3:1, the active biocatalyst isolated from the yeast. Comparative analysis of their tryptic peptides by electrospray tandem mass spectrometry allowed unequivocal assignment of the modification as the oxidation of Cys108 into cysteine sulfinic acid. Cys108 is likely located on a surface-exposed protein region within the FAD binding domain, but remote from the active center. Its oxidized side chain was remarkably stable in solution, thus enabling the relative biochemical characterization of native and modified enzyme forms. The oxidation of Cys108 causes a global conformational response that affects the protein environment of the FAD cofactor. In comparison with the native enzyme, it results in a 4-fold decreased specific activity, reflecting a catalytic efficiency for reduction of dioxygen lowered by about the same factor ; and markedly decreased propensity to aggregate under conditions of thermal denaturation. These results open up unprecedented routes for stabilization of the oxidase, and underscore the possible significance of protein chemical heterogeneity for biocatalyst function and stability.

Izvorni jezik
Engleski

Znanstvena područja
Biotehnologija



POVEZANOST RADA


Projekti:
0058011

Ustanove:
Prehrambeno-biotehnološki fakultet, Zagreb

Profili:

Avatar Url Anita Slavica (autor)


Citiraj ovu publikaciju

Slavica, Anita; Dib, Iskandar; Nidetzky, Bernd
Single-site oxidation, cysteine 108 to cysteine sulfinic acid, in D-amino acid oxidase from Trigonopsis variabilis and its structural and functional consequences // Applied and Environmental Microbiology, 71 (2005), 12; 8061-8068 (međunarodna recenzija, članak, znanstveni)
Slavica, A., Dib, I. & Nidetzky, B. (2005) Single-site oxidation, cysteine 108 to cysteine sulfinic acid, in D-amino acid oxidase from Trigonopsis variabilis and its structural and functional consequences. Applied and Environmental Microbiology, 71 (12), 8061-8068.
@article{article, year = {2005}, pages = {8061-8068}, keywords = {enzyme activity, oxidative modification, cysteine sulfinic acid, MS}, journal = {Applied and Environmental Microbiology}, volume = {71}, number = {12}, issn = {0099-2240}, title = {Single-site oxidation, cysteine 108 to cysteine sulfinic acid, in D-amino acid oxidase from Trigonopsis variabilis and its structural and functional consequences}, keyword = {enzyme activity, oxidative modification, cysteine sulfinic acid, MS} }
@article{article, year = {2005}, pages = {8061-8068}, keywords = {enzyme activity, oxidative modification, cysteine sulfinic acid, MS}, journal = {Applied and Environmental Microbiology}, volume = {71}, number = {12}, issn = {0099-2240}, title = {Single-site oxidation, cysteine 108 to cysteine sulfinic acid, in D-amino acid oxidase from Trigonopsis variabilis and its structural and functional consequences}, keyword = {enzyme activity, oxidative modification, cysteine sulfinic acid, MS} }

Časopis indeksira:


  • Current Contents Connect (CCC)
  • Web of Science Core Collection (WoSCC)
    • Science Citation Index Expanded (SCI-EXP)
    • SCI-EXP, SSCI i/ili A&HCI
  • Scopus
  • MEDLINE





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