Purification and characterisation of phenoloxidase from amphioxus Branchiostoma belcheri tsingtauense

Pang, Qiuxiang; Zhang, Shicui; Shi, Xiaodong; Su, Feng; Wu, Di

doi:10.1016/j.fsi.2004.12.001

Cited by 27 publications

(17 citation statements)

References 46 publications

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“…PO in the humoral fluids from unchallenged and challenged amphioxus both comprises three bands, with a molecular mass of 72, 46 and 44 kDa, respectively, confirming our previous observation [15]. Notably, band 2 was susceptible to challenge with V. alginolyticus or E. coli, while bands 1 and 3 were not ( Fig.…”

supporting

confidence: 89%

“…It has been previously shown that the humoral fluids including the blood in amphioxus contain PO activity [14,15]. Whether microbial challenge induces changes in PO level in the humoral fluids in amphioxus is not yet known.…”

Section: Introductionmentioning

confidence: 99%

“…The non-reducing SDS-PAGE was run at 120 V for 2 h, and the molecular mass markers were myosin (200 kDa), calmodulin binding protein (130 kDa), rabbit phosphorylase B (97.4 kDa), bovine serum albumin (66.2 kDa), and rabbit actin (43 kDa). PO activity on the polyacrylamide gels was detected according to the method of Nellaiappan and Vinayagam [19] with slight modifications [15]. The gels were washed twice in ice-cold distilled water, and incubated in 50 mM PBS (pH 6.5) containing 10 mM 4-methylcatechol and 0.3% MBTH dissolved in 25% ethanol.…”

mentioning

confidence: 99%

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Induction of phenoloxidases in the humoral fluids of amphioxus Branchiostoma belcheri by Vibrio alginolyticus and Escherichia coli

Pang¹,

Zhang²,

Zhao³

2009

Fish & Shellfish Immunology

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supporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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Induction of phenoloxidases in the humoral fluids of amphioxus Branchiostoma belcheri by Vibrio alginolyticus and Escherichia coli

Pang¹,

Zhang²,

Zhao³

2009

Fish & Shellfish Immunology

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“…In most of these applications, laccases are used together with a redox mediator. At the end of 2005 three industrial processes were using laccases: dye bleaching, lignin bleaching and bleaching of cork for bottled wine [19].…”

Section: Introductionmentioning

confidence: 99%

Improvement of Laccase Production in Pluerotus pulmonarius-LAU 09 by Mutation

Adebayo¹,

Kola²,

Yadav³

et al. 2012

MICROBIOLOGY

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The strain improvement of P. pulmonarius LAU 09 for laccase production was carried out by UV-light exposure at 210nm (UV-sterilizer, Millipore xx63 70000) for 90minutes. The highest laccase activity of 2.5 Uml-1 was produced by LAU 90 (mutant) in comparison with obtained yield by wild type LAU 09 (1.75 Uml-1). The optimal physiological parameters for optimum laccase production by LAU 90 are 30OC of temperature, pH of 5.0 and incubation of 168hours. The mutant strain gave the highest total enzyme activity and enzyme specific activity of 71.7 x 10-4 mol/min and 89.63X 10-4 IU/mg of purified laccase enzyme respectively, with molecular weight of 97KDa. The activity of the purified enzyme was strongly inhibited by Sodium azide and EDTA at 1ml/0.1mM and 0.5ml/100mM respectively. The result obtained shown an improved performance of the mutant (LAU 90) strain of P. pulmonarius in laccase production over the wild (LAU 09) type.

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“…The method of Pang et al 12 was used to purify PO of E. kuehniella in a three-step procedure as follows:…”

Section: Purification Of Pomentioning

confidence: 99%

Purification and characterization of a phenoloxidase in the hemocytes ofEphestia kuehniellaZeller (Lepidoptera: Pyralidae): effects of insect growth regulators and endogenous inhibitors

Delkash-Roudsari

Zibaee

Bigham

2014

Journal of Enzyme Inhibition and Medicinal Chemistry

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A phenoloxidase was extracted and purified from hemocytes of Ephestia kuehniella by using ammonium sulfate, Sepharyl G-100 and DEAE-Cellulose fast flow chromatographies. At the final stage of purification, a protein was purified by molecular mass of 78.5 kDa, specific activity of decreased activity of the purified PO but Ca 2+ and Cu 2+ increased the enzymatic activity. EDTA (General chelating agent), DTC (Copper chelating agent) and EGTA (Calcium chelating agent) significantly decreased PO activity but TTHA (Magnesium chelating agent) showed no statistically significant effects. Kinetic parameters of the purified enzyme showed the highest V max when L-DOPA was used as substrate but no significant differences were observed in case of K m for used L-DOPA, pyrocatechol and hydroquinone. In vitro inhibition of the purified PO by using two insect growth regulators, Hexaflumuron and Pyriproxyfen, revealed IC 50 of 96.41 and 38.59 mg/ml for these compounds, respectively. Kinetic studies using different concentrations of L-DOPA and IC 50 concentrations of the two IGRs revealed the increase of K m value versus control and competitive inhibition. Finally, column chromatography of hemolymph revealed peak III showing endogenous inhibitors of phenoloxidase by molecular weight of 27.3 that showed competitive inhibition on the PO.

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Purification and characterisation of phenoloxidase from amphioxus Branchiostoma belcheri tsingtauense

Cited by 27 publications

References 46 publications

Induction of phenoloxidases in the humoral fluids of amphioxus Branchiostoma belcheri by Vibrio alginolyticus and Escherichia coli

Induction of phenoloxidases in the humoral fluids of amphioxus Branchiostoma belcheri by Vibrio alginolyticus and Escherichia coli

Improvement of Laccase Production in Pluerotus pulmonarius-LAU 09 by Mutation

Purification and characterization of a phenoloxidase in the hemocytes ofEphestia kuehniellaZeller (Lepidoptera: Pyralidae): effects of insect growth regulators and endogenous inhibitors

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