Gel electrophoresis of polyphenol oxidase with instant identification by in situ blotting

Cheng, Tsai Mu; Huang, Po-Chiun; Pan, Ju Pin; Lin, Kuan Yu; Mao, Simon J.T.

doi:10.1016/j.jchromb.2006.08.046

Cited by 12 publications

(9 citation statements)

References 30 publications

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“…Indeed, PPO as well as the inhibitor would be expected to be present in the extract of cauliflower floret. Apart from that, gel electrophoretic analysis also revealed reactivity toward 4-MC was not exclusively linked to a single protein band, which might indicate the existence of various enzyme isoforms of PPO, as observed before in unpurified protein extracts of potato tubers , and other biological materials. , …”

Section: Discussionsupporting

confidence: 56%

“…On the other hand, gel electrophoretic analysis revealed high reactivity of some extracted proteins toward 4-MC, despite spectrophotometric measurements being below the detection limit. Conflicting results between a conventional spectrophotometric PPO assay and a PAGE blot have been reported before and were explained by an endogenous PPO inhibitor present in the extract . The presence of such inhibitors (or activators) in the protein extract most likely does not affect the activity observed by means of PAGE .…”

Section: Discussionmentioning

confidence: 87%

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Polyphenol Oxidase Containing Sidestreams as Emulsifiers of Rumen Bypass Linseed Oil Emulsions: Interfacial Characterization and Efficacy of Protection against in Vitro Ruminal Biohydrogenation

Gadeyne

Neve

Vlaeminck

et al. 2016

J. Agric. Food Chem.

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The low transfer in ruminants of dietary polyunsaturated fatty acids to the milk or peripheral tissues is largely due to ruminal biohydrogenation. Lipids emulsified by a polyphenol oxidase (PPO) rich protein extract of red clover were shown before to be protected against this breakdown after cross-linking with 4-methylcatechol. Protein extracts of 13 other vegetal resources were tested. Surprisingly, the effectiveness to protect emulsified lipids against in vitro ruminal biohydrogenation largely depended on the origin of the extract and its protein concentration but was not related to PPO activity. Moreover, PPO isoforms in vegetal sources, effectively protecting emulsified lipids, were diverse and their presence at the emulsion interface did not seem essential. Potato tuber peels were identified as an interesting biological source of emulsifying proteins and PPO, particularly since protein extracts of industrial potato sidestreams proved to be suitable for the current application.

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Section: Discussionsupporting

confidence: 56%

Section: Discussionmentioning

confidence: 87%

Polyphenol Oxidase Containing Sidestreams as Emulsifiers of Rumen Bypass Linseed Oil Emulsions: Interfacial Characterization and Efficacy of Protection against in Vitro Ruminal Biohydrogenation

Gadeyne

Neve

Vlaeminck

et al. 2016

J. Agric. Food Chem.

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“…The molecular weight of PPO varies, however, between species [21]. In the present work, the purified LsPPO migrated as a band of approximately 65 kDa (data not shown), upon SDS -polyacrylamide gel electrophoresis.…”

Section: Extraction and Purification Of Lsppomentioning

confidence: 40%

Differential in vitro inhibition of polyphenoloxidase from a wild edible mushroom Lactarius salmonicolor

Dedeoğlu

Guler

2009

Journal of Enzyme Inhibition and Medicinal Chemistry

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The polyphenol oxidase (LsPPO) from a wild edible mushroom Lactarius salmonicolor was purified using a Sepharose 4B-Ltyrosine-p-amino benzoic acid affinity column. At the optimum pH and temperature, the K M and V Max values of LsPPO towards catechol, 4-methylcatechol and pyrogallol were determined as 0.025 M & 0.748 EU/mL, 1.809 £ 10 23 M & 0.723 EU/mL and 9.465 £ 10 23 M & 0.722 EU/mL, respectively. Optimum pH and temperature values of LsPPO for the three substrates above ranged between the pH 4.5-11.0 and 5-508C. Enzyme activity decreased due to heat denaturation with increasing temperature. Effects of a variety of classical PPO inhibitors were investigated opon the activity of LsPPO using catechol as the substrate. IC 50 values for glutathione, p-aminobenzenesulfonamide, L-cysteine, L-tyrosine, oxalic acid, b-mercaptoethanol and syringic acid were determined as 9.1 £ 10 24 , 2.3 £ 10 24 M, 1.5 £ 10 24 M, 3.8 £ 10 27 M, 1.2 £ 10 24 M, 4.9 £ 10 24 M, and 4 £ 10 24 M respectively. Thus L-tyrosine was by far the most effective inhibitor. Interestingly, sulfosalicylic acid behaved as an activator of LsPPO in this study.

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“…Polyphenol oxidase (PPO) activity was determined using a modified method of Cheng et al . (2007) and Ni Eidhin et al . (2010).…”

Section: Methodsmentioning

confidence: 96%

Improved solubility of proteins from white and red clover – inhibition of redox enzymes

Amer

Juul

Møller

et al. 2020

Int J of Food Sci Tech

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New alternative protein sources are needed. Clover grasses as white clover (WC) and red clover (RC) may provide a novel protein source to achieve high-quality food protein. To prevent protein oxidation, endogenous oxidative enzymes as polyphenol oxidase (PPO) and peroxidases were inhibited with sulphite. RC showed higher PPO activity than WC. Low sulphite inhibited the PPO activity in both species, but browning still occurred in RC. Sulphite did not affect the polyphenol (PP) content in WC, rather suggesting peroxidase than PPO activity. In RC juice, the PP content increased in a dose-dependent manner with increasing sulphite. High sulphite impaired browning and increased the content of soluble rubisco in RC, mainly by increasing the content of native rubisco. In vitro digestibility of RC protein increased with increasing sulphite. In conclusion, sulphite inhibited oxidative enzymes and increased the quality of protein extracted from WC and RC.

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Gel electrophoresis of polyphenol oxidase with instant identification by in situ blotting

Cited by 12 publications

References 30 publications

Polyphenol Oxidase Containing Sidestreams as Emulsifiers of Rumen Bypass Linseed Oil Emulsions: Interfacial Characterization and Efficacy of Protection against in Vitro Ruminal Biohydrogenation

Polyphenol Oxidase Containing Sidestreams as Emulsifiers of Rumen Bypass Linseed Oil Emulsions: Interfacial Characterization and Efficacy of Protection against in Vitro Ruminal Biohydrogenation

Differential in vitro inhibition of polyphenoloxidase from a wild edible mushroom Lactarius salmonicolor

Improved solubility of proteins from white and red clover – inhibition of redox enzymes

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