Isolation and properties of L-lysine-α-oxidase from the fungus Trichoderma cf. aureoviride RIFAI VKM F-4268D

L-lysine-α-oxidase (LO) synthesized by the fungus Trichoderma cf. aureoviride Rifai VKM F-4268D under salt stress conditions was isolated and characterized. The newly developed method for the isolation and purification of the enzyme was based on its precipitation from the culture liquid by copper sulfate. The subsequent LO purification by the methods of hydrophobic (Octyl Sepharose) and ion exchange (DEAE ToyoPearl) chromatography yielded a homogeneous enzyme preparation with a high degree of purification (310-fold) and high specific activity (90 U/mg protein). The molecular mass of the enzyme determined by gel filtration and native electrophoresis was 115-116 kDa. According to the data of SDS electrophoresis, LO was a dimer with identical subunits (57-58 kDa). The optical absorption spectrum of LO corresponded to the flavoprotein spectrum with maximums at 278, 390, and 465 (a shoulder at 490) nm. LO is a stereospecific enzyme oxidizing almost exclusively L-lysine (pH optimum 7.8-8.2). Insignificant activity was observed against L-ornithine and L-arginine. LO was shown to be stable at temperatures up to 50°C. © 2012 Pleiades Publishing, Ltd.

Authors
Arinbasarova A.Y.1 , Ashin V.V.1 , Makrushin K.V.1 , Medentsev A.G.1 , Lukasheva E.V. 2 , Berezov T.T. 2
Number of issue
5
Language
English
Pages
549-554
Status
Published
Volume
81
Year
2012
Organizations
  • 1 Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, pr. Nauki 5, Pushchino, Moscow oblast 142290, Russian Federation
  • 2 Peoples' Friendship University of Russia, ul. Miklukho-Maklaya 6, Moscow 117198, Russian Federation
Keywords
biosynthesis; fungus Trichoderma aureoviride; isolation; L-lysine-α-oxidase; purification; substrate specificity
Date of creation
19.10.2018
Date of change
19.10.2018
Short link
https://repository.rudn.ru/en/records/article/record/2254/
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