Novel affinity purification of xanthine oxidase from Arthrobacter M3

Xanthine oxidase (XO) is an important enzyme catalyzing the hydroxylation of hypoxanthine to xanthine and xanthine to uric acid which is excreted by kidneys. Excessive production and/or inadequate excretion of uric acid results in hyperuricemia. This paper presents a detailed review of methods of isolation, determination of xanthine oxidase activity, and the effect of plant extracts and their constituents on it. Determining the content and activities of XO can be used for diagnostic purposes. Testing inhibition of XO is important for detection of potentially effective compounds or extracts that can be used to treat diseases that are caused by increased activity of XO.In vitrobioassays are used to examine test material for XO inhibition, as inhibitors of XO may be potentially useful for the treatment of gout or other XO induced diseases. Several authors reported on the XO inhibitory potential of traditionally used medicinal plants.

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“…Zhang et al [58] isolated and purified XO from Arthrobacter M3. The Arthrobacter M3 culture was transferred into induction medium.…”

Section: Isolation and Purification Of Xomentioning

confidence: 99%

Xanthine Oxidase: Isolation, Assays of Activity, and Inhibition

Kostić

Dimitrijević

Stojanović

et al. 2015

Journal of Chemistry

148

109

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“…After chromatography on the Sephacryl S-300 column, SLXO was purified 31.8-fold with a specific activity of 3.5 units / mg protein and 24.1% recovery (Table 1). Buffalo milk xanthine oxidase; 10.66 unit/mg [28], bovine milk xanthine oxidase; 1.086 unit/mg [24], 0.3 unit/mg [29] and 15.8 unit/mg [30], human liver xanthine oxidase; 0.00061 unit/mg [31], and 0.036 unit/mg [32], buffalo liver xanthine oxidase; 7.2 unit/mg [33], rat liver xanthine oxidase; 14 unit/mg [27] and Arthrobacter M3 was 8.6 unit/mg [13]. Electrophoretic analysis of n-butanol extract, DEAE-cellulose fraction and Sephacryl S-300 purified fraction of SLXO on 7 % native PAGE revealed single protein band corresponded the enzyme activity band of the purified xanthine oxidase enzyme (Fig.…”

Section: Purification Of Sheep Liver Xanthine Oxidasementioning

confidence: 99%

“…It has been proposed that xanthine dehydrogenase in the endothelial cell cytoplasm is released following endothelial injury by microbes, and is converted to the oxidase form in the oxygen rich environment of blood. Xanthine oxidase can oxidize substrates in the blood, leading to the formation of oxygen derived free radicals which have been suggested to provide oxidative defence [12,13]. One of the most important xanthine oxidase competitive inhibitors is allopurinol that is a substrate analog, has similar chemical structure to hypoxanthine and used in treatment of hyperuricemia and gout [1,14].…”

Section: Introductionmentioning

confidence: 99%

Purification and Characterization of Xanthine Oxidase from Liver of the Sheep (Ovis Aries)

Zaahkouk

Darwish

Masoud

et al. 2019

JAA

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Xanthine oxidase is a commercially important enzyme with wide area of medical applications to develop diagnostic kits. Xanthine oxidase was extracted, purified and characterized from sheep liver (SLXO). The purification procedure involved acetone precipitation and chromatography on DEAE-cellulose and Sephacryl S-300 columns. The sheep liver xanthine oxidase was homogeneously purified 31.8 folds with 3.5 U/mg specific activity and 24.1% recovery. SLXO native molecular weight was 150 kDa and on SDS-PAGE appeared as single major band of 75 kDa representing a homodimer protein. Isoelectric focusing of the purified SLXO resolved into two closely related isoforms with pI values of 5.6 and 5.8. The apparent Km for xanthine oxidase at optimum pH 7.6 was found to be 0.9 mM xanthine. FeCl2 and NiCl2 increased the activity of SLXO, while CuCl2 and ZnCl2 were found to be potent inhibitors of the purified enzyme. Allopurinol inhibits SLXO competitively with one binding site on the purified molecule and Ki value of 0.06 mM.

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“…Xanthine oxidase from various sources are proteins of molecular weight *300 kDa and mainly contain molybdopterin cofactors [2,8]. Each enzymatic unit contains flavin adenine dinucleotide, molybdenum and iron sulfur centres [2].…”

Section: Introductionmentioning

confidence: 99%

“…The purified enzyme is used in the assay method of super oxide dismutase [8]. Xanthine oxidase also has role in the food industries for assessment of freshness of meat [9].…”

Section: Introductionmentioning

confidence: 99%

Thermostable Xanthine Oxidase Activity from Bacillus pumilus RL-2d Isolated from Manikaran Thermal Spring: Production and Characterization

Sharma

Thakur

et al. 2015

Indian J Microbiol

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Xanthine oxidase is an important enzyme of purine metabolism that catalyzes the hydroxylation of hypoxanthine to xanthine and then xanthine to uric acid. A thermostable xanthine oxidase is being reported from a thermophilic organism RL-2d isolated from the Manikaran (Kullu) hot spring of Himachal Pradesh (India). Based on the morphology, physiological tests, and 16S rDNA gene sequence, RL-2d was identified as Bacillus pumilus. Optimization of physiochemical parameters resulted into 4.1-fold increase in the xanthine oxidase activity from 0.051 U/mg dcw (dry cell weight) to 0.209 U/mg dcw. The xanthine oxidase of B. pumilus RL-2d has exhibited very good thermostability and its t 1/2 at 70 and 80°C were 5 and 1 h, respectively. Activity of this enzyme was strongly inhibited by Hg 2? , Ag ? and allopurinol. The investigation showed that B. pumilus RL-2d exhibited highest xanthine oxidase activity and remarkable thermostability among the other xanthine oxidases reported so far.

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Novel affinity purification of xanthine oxidase from Arthrobacter M3

Cited by 9 publications

References 13 publications

Xanthine Oxidase: Isolation, Assays of Activity, and Inhibition

Xanthine Oxidase: Isolation, Assays of Activity, and Inhibition

Purification and Characterization of Xanthine Oxidase from Liver of the Sheep (Ovis Aries)

Thermostable Xanthine Oxidase Activity from Bacillus pumilus RL-2d Isolated from Manikaran Thermal Spring: Production and Characterization

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