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Structure and activity of a novel robust peroxidase from Alkanna frigida cell culture.
Soleimani Asl, Saeed; Karkhane, Ali Asghar; Zamani Amirzakaria, Javad; Akbari Noghabi, Kambiz; Ghashghaie, Jaleh; Haghbeen, Kamahldin.
Afiliación
  • Soleimani Asl S; National Institute for Genetic Engineering and Biotechnology, Tehran, Iran.
  • Karkhane AA; National Institute for Genetic Engineering and Biotechnology, Tehran, Iran.
  • Zamani Amirzakaria J; National Institute for Genetic Engineering and Biotechnology, Tehran, Iran.
  • Akbari Noghabi K; National Institute for Genetic Engineering and Biotechnology, Tehran, Iran.
  • Ghashghaie J; Laboratoire d'Ecologie, Systématique et Evolution (ESE), Université de Paris-Sud, CNRS, AgroParisTech, Université de Paris-Saclay, 91400 Orsay, France.
  • Haghbeen K; National Institute for Genetic Engineering and Biotechnology, Tehran, Iran. Electronic address: Kamahl@nigeb.ac.ir.
Phytochemistry ; 194: 113022, 2022 Feb.
Article en En | MEDLINE | ID: mdl-34826793
Demands for peroxidases (POX)s with diverse physicochemical properties have steadily grown as more applications of POXs are demonstrated. Plants are among the best sources of versatile POXs, and plant biotechnology, as an agricultural hassle-free technology, promises to circumvent the limitations of natural resource exploitation and to address the demands. Following this trend, it was shown that POX production steadily increased during the 31-day subculture of Alkanna frigida (from Boraginaceae) callus on Murashige-Skoog medium containing 2,4-dichlorophenoxyacetic acid (10-6 M) and kinetin (10-5 M). The purified cationic enzyme (POXalf) maintained its optimal activity over pH 4-7 for 2 years. It was resistant to H2O2 high concentrations (IC50 = 543.7 mM) and showed high specific activity in the reaction with phenol (4320.5 AU mg-1 > 20-fold of HRP AU). Furthermore, the specificity constant ratio of guaiacol to phenol indicated a 100 times faster reaction of POXalf with guaiacol. However, in contrast to HRP, it had little effect on diazo derivatives of aniline and meta-diaminobenzene. Based on the resulting primary structure from the tandem mass analysis, the POXalf 3D structure was constructed via homology modelling. Despite the high topological similarity between the HRP and POXalf structures, there were important differences between the active site pockets that could explain the observed differences in the corresponding substrate spectra and the specific activities. Considering the dynamics of POXalf production, its inactivity towards IAA and its high affinity for guaiacol, POXalf may have associated roles with A. frigida cell wall construction and monolignol metabolism.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Peroxidasa / Boraginaceae Idioma: En Revista: Phytochemistry Año: 2022 Tipo del documento: Article País de afiliación: Irán

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Peroxidasa / Boraginaceae Idioma: En Revista: Phytochemistry Año: 2022 Tipo del documento: Article País de afiliación: Irán
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