Properties of Rhodotorula gracilis d-Amino Acid Oxidase Immobilized on Magnetic Beads through His-Tag

Kuan, I-Ching; Liao, Renjie; Hsieh, Hao‐Chieh; Chen, Kuanchun; Yu, Chih-Wen

doi:10.1263/jbb.105.110

Cited by 24 publications

(16 citation statements)

References 27 publications

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“…DAO activity was determined by measuring an increase in absorbance accompanying the oxidation of o-phenylenediamine, which was catalyzed by horseradish peroxidase using H 2 O 2 released from the DAO reaction. To study the oxidative stability against H 2 O 2 , a different assay measuring pyruvate production as performed by Kuan et al (2008) was used to avoid the influence from exogenous H 2 O 2 . Assays were performed in triplicate.…”

Section: Dao Activity Assaymentioning

confidence: 99%

Stabilization of d-amino acid oxidase from Rhodosporidium toruloides by immobilization onto magnetic nanoparticles

et al. 2008

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D-amino acid oxidase from Rhodosporidium toruloides was immobilized onto glutaraldehyde-activated magnetic nanoparticles. Approximately four enzyme molecules were attached to one magnetic nanoparticle when the weight ratio of the enzyme to the support was 0.12. After immobilization, the T(m) was increased from 45 degrees C of the free form to 55 degrees C. In the presence of 20 mM H2O2, the immobilized form retained 93% of its activity after 5 h while the free form was completely inactivated after 3.5 h.

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Section: Dao Activity Assaymentioning

confidence: 99%

Stabilization of d-amino acid oxidase from Rhodosporidium toruloides by immobilization onto magnetic nanoparticles

et al. 2008

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“…The tested fungi have been chosen because of their industrial and/or medical importance. Rhodotorula strains are known for the intracellular production of useful amino acid oxidases (Kuan et al 2008) and β-carotene (Vijayalakshmi et al 2001). Secondary metabolites produced inside the cells of A. fumigatus and P. citrinum are also of commercial interest (Du et al 2010; Chen et al 2011; Malmstrom et al 2000; Lim et al 2007; Singh et al 2011).…”

Section: Discussionmentioning

confidence: 99%

Comparative study of fungal cell disruption—scope and limitations of the methods

et al. 2011

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Simple and effective protocols of cell wall disruption were elaborated for tested fungal strains: Penicillium citrinum, Aspergillus fumigatus, Rhodotorula gracilis. Several techniques of cell wall disintegration were studied, including ultrasound disintegration, homogenization in bead mill, application of chemicals of various types, and osmotic shock. The release of proteins from fungal cells and the activity of a cytosolic enzyme, glucose-6-phosphate dehydrogenase, in the crude extracts were assayed to determine and compare the efficacy of each method. The presented studies allowed adjusting the particular method to a particular strain. The mechanical methods of disintegration appeared to be the most effective for the disintegration of yeast, R. gracilis, and filamentous fungi, A. fumigatus and P. citrinum. Ultrasonication and bead milling led to obtaining fungal cell-free extracts containing high concentrations of soluble proteins and active glucose-6-phosphate dehydrogenase systems.

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“…One unit of enzyme activity was defined as the amount of enzyme that produces 1 µmol of pyruvate per minute as per the assay conditions. D-aao activity (o-phenelenediamine method) was determined spectrophotometrically by measuring the absorbance increase accompanying the oxidation of o-phenylenediamine [25]. A 1 ml assay mixture contained 30 mM D-alanine, 0.03% (w/v) o-phenylenediamine, 5 U/ml horseradish peroxidase (HiMedia, India) and 0.3 g PAnisodium alginate-D-aao beads in 0.1 M sod.…”

Section: D-aaomentioning

confidence: 99%

Docking of Polyaniline with D-Amino Acid Oxidase of Rhodosporidium toruloides and Pig Kidney-An Insight into the Mechanism of Binding for Immobilization in Polyaniline Supports

Singh¹,

Buragohain²

2014

J Microb Biochem Technol

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The interaction of Polyaniline (PAni) with Rhodosporidium toruloides D-amino acid oxidase (RtDaao) and pig kidney D-aao (PkDaao) was studied by bioinformatics approach. The interaction of PAni with RtDaao does not interfere with the substrate binding and hence the D-amino acid oxidase (D-aao) activity is unaffected by the ligand. The active site cavity of pig kidney D-aao (PkDaao) is comprised of the residues Leu 51, Tyr 224, Tyr 228, Arg 283 and Gly 313. PAni interacts with Leu 51 and Thr 317 of chain G of the PkDaao with interaction energy of -2.5 and -1.09 kcal/mol respectively. D-aao was immobilized onto PAni-sodium alginate beads using glutaraldehyde as the crosslinking agent. 1g of the PAni-sodium alginate D-aao beads contained 0.093 ml or 0.385 U of the D-aao enzyme. The Activity Yield (AY) was calculated as 18.92% as determined by the pyruvate method of detection while, the AY was calculated as 17.3% as determined by the o-PDA method of D-aao assay of the beads. The PAni-sodium alginate D-aao beads were characterized by Fourier Transform Infrared spectroscopy (FTIR), X-ray Diffraction (XRD), Energy Dispersive X-ray Diffraction (EDX), Thermogravimetric Analysis (TGA) and Scanning Electron Microscopic analysis.

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Properties of Rhodotorula gracilis d-Amino Acid Oxidase Immobilized on Magnetic Beads through His-Tag

Cited by 24 publications

References 27 publications

Stabilization of d-amino acid oxidase from Rhodosporidium toruloides by immobilization onto magnetic nanoparticles

Stabilization of d-amino acid oxidase from Rhodosporidium toruloides by immobilization onto magnetic nanoparticles

Comparative study of fungal cell disruption—scope and limitations of the methods

Docking of Polyaniline with D-Amino Acid Oxidase of Rhodosporidium toruloides and Pig Kidney-An Insight into the Mechanism of Binding for Immobilization in Polyaniline Supports

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