Purification and Partial Biochemical Characterization of Polyphenol Oxidase from Mango (<i>Mangifera indica</i> cv. Manila)

Palma-Orozco, Gisela; Marrufo-Hernández, Norma A.; Sampedro, José G.; Nájera, Hugo

doi:10.1021/jf5029784

Cited by 34 publications

(46 citation statements)

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“…The best chromatography combination to our knowledge is hydroxyapatite column followed by a gel filtration column used for banana PPO purification where authors report 635.7‐fold purification, yield of 3%, protein content of 0.9 mg, and specific activity of 219.3 μkat mg −1 (Yang, Fujita, Ashrafuzzaman, Nakamura, & Hayashi, ). The same type of chromatographic separation used in this work (HIC followed by IEC) was reported for PPO from mamey (Palma‐Orozco et al, ) and Manila mango (Palma‐Orozco et al, ); having higher yields 11 and 3%, respectively, lower activity increases (183‐fold for mamey and 216‐fold for mango), and similar protein content than obtained herein.…”

Section: Resultssupporting

confidence: 53%

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Purification, partial biochemical characterization and inactivation of polyphenol oxidase from Mexican Golden Delicious apple (Malus domestica )

Marrufo-Hernández

Palma-Orozco

Beltrán

et al. 2017

Journal of Food Biochemistry

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Polyphenol oxidase (PPO) is a critical molecule because of its participation in enzymatic browning in fruits and vegetables during ripening, handling, storage, and processing. In this contribution, PPO was extracted, purified, and characterized from Golden Delicious apples, PPO activity was increased 319‐fold with a yield of 1.6%. A monomer of 58 kDa was obtained by gel filtration and SDS‐PAGE; the optimum temperature and pH were 35°C and 6.0, respectively; the best catalytic efficiencies (Vmax/Km ratios [U/(mg·mM)]) were 15,402 for 4‐methylcatechol and 5,941 for pyrogallol. The most efficient inhibitors were sodium metabisulfite (≥1 mM) and tropolone (≥10 mM). PPO activity decreased as the temperature increased and thermal inactivation was achieved at 80°C in 5 min. An Ea of 69.2 kJ/mol was estimated from the kinetic inactivation at different temperatures. Microwave irradiation inactivates PPO after 120 s in crude extract and after 60 s in fresh apple juice. Practical applications According to FAO (Food and Agricultural Organization) data, Mexico produced about 858,608 tons of apple in 2013, Mexico is in 30th place in the world overall apple production. It is important to characterize the Polyphenol Oxidase (PPO) from Golden Delicious apple and provide data on its inactivation. PPO has been considered a crucial molecule because of its participation in enzymatic browning in several fruit and vegetables during ripening, handling, storage, and processing, being responsible for economical loses for producers and industry. In this contribution PPO was extracted, purified, and partially characterized from Mexican Golden Delicious apples by fast protein liquid chromatography. Also some inactivation methods like the use of several inhibitors and antibrowning agents, thermal and microwave inactivation where tested. The information obtained in this research is important for better understanding of PPO from Golden Delicious apple and gives information on ways to optimize inactivation process of the purified enzyme or in apple juice.

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Section: Resultssupporting

confidence: 53%

“…The dialyzed solution added with Aprotinin (0.033 mg L −1 ) was loaded onto a previously equilibrated Mono Q HR 10/10 column to carry on ion exchange chromatography (IEC). Proteins were eluted by a linear increase in the ionic strength collecting fractions of 2 mL, PPO activity and protein content were measured (Palma‐Orozco et al, ).…”

Section: Methodsmentioning

confidence: 99%

Purification, partial biochemical characterization and inactivation of polyphenol oxidase from Mexican Golden Delicious apple (Malus domestica )

Marrufo-Hernández

Palma-Orozco

Beltrán

et al. 2017

Journal of Food Biochemistry

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“…Triton X‐100, a nonionic detergent, is widely used in the extraction of PPO as it provides a mild nondenaturing condition for proteins. In addition, PVPP and ascorbic acid are included in the extraction buffer to minimize o‐quinone formation and prevent phenol‐protein interactions (Palma‐Orozco, Marrufo‐Hernández, Sampedro, & Nájera, ). The characteristic absorption peak of the enzyme‐substrate mixture was determined to be 413nm (Figure ), which was different from previously reported 410nm of Fuji apple PPO (Liu, Zhao, Gan, & Ni, ).…”

Section: Resultsmentioning

confidence: 99%

Inhibition kinetics of oligochitosan as an uncompetitive inhibitor on fuji apple polyphenol oxidase

Guo

Zhang

et al. 2018

J Food Biochem

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This study aimed to investigate the effect of oligochitosan on purified polyphenol oxidase (PPO) from Fuji apple and the associated mechanisms. PPO was firstly separated and purified in a four‐step procedure and the molecular weight was estimated to be about 65 kDa by SDS‐PAGE. PPO showed an optimum activity at pH 7 and 35°C, and it was stable in the temperature range of 25–55°C with more than 50% residual activity. Thermal inactivation parameters indicated that the Fuji apple PPO was very heat‐labile. Using catechol as the substrate, the PPO activity decreased with the increase of oligochitosan concentration, and only 30% activity remained when the concentration reached 4mg/mL. Oligochitosan showed uncompetitive type reversible inhibition with Ki value of 0.26mg/mL. Data obtained in this study provided theoretical basis for controlling enzymatic browning during apple processing with oligochitosan. Practical applications Processing operations, such as peeling, cutting, and juicing have the same problem: enzymatic browning, which causes a decrease in market value and economic loss. The most common approach for preventing browning is using anti‐browning agents, however, consumers are demanding a reduction in the overall use of chemicals on foods safe and asking effective substitutes especially natural inhibitors with the increased health awareness. This research systematically characterizes the enzymatic browning related PPO from apple and demonstrated that oligochitosan can inhibited PPO activity as an uncompetitive inhibitor, which will help in the development of oligochitosan to be a new type of anti‐browning agent.

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“…Km values were found to vary in apricot (Arslan et al, 1998), Hemşin apple (Aydin et al, 2015), mamey (Palma-Drozco et al, 2014), Chinese Toon (Wang et al, 2013) and quince (Yagar & Sagıroglu, 2002) as 6.6 mM, 6.8 mM, 44 mM, 10.059 mM and 4.54 mM, respectively. When using 4-methylcatechol as the substrate for PPD, Km values were calculated as 3.14 mM in mango (Palma-Drozco et al, 2014) and 18.2 mM in Jackfruit (Tao et al, 2013). Moreover, when using L-Dopa as the substrate, a Km value of 17.8 mM was reported in quince (Yagar & Sagıroglu, 2002).…”

Section: Kinetic Characteristics Of Ppo Using Different Substratesmentioning

confidence: 97%

“…The molecular weights of the PPD from various plants were reported as 112 kDa for eggplant (Mishra et al, 2012), 31.5 kDa for mango (Palma-Drozco et al, 2014), multiple bands of 53, 112 and 144 kDa for Ataulfo mango (Cheema & Sommerhalter, 2015), 82 kDa for atemoya fruit (Chaves et al, 2011).…”

Section: Polyacrylamide Gel Electrophoresismentioning

confidence: 99%

Purification and characterization of polyphenol oxidase from purslane

Güven

Bekler

et al. 2017

Food Sci. Technol

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The polyphenol oxidase (PPD) is an enzyme that is responsible for the enzymatic browning of fruits and vegetables. This is generally undesired process and need to be prevented in food technology. PPD from purslane was purified, characterised and the kinetic parameters for three substrates namely, catechol, L-Dopa and 4-methylcatechol were determined. The optimum pH and temperature values were found to be pH 7.0 and 50 °C, respectively using the catechol as substrate. The apparent molecular weight of the PPD from purslane was determined as high as 163 kDa by partially denaturing SDS-PAGE. Moreover, the inhibition kinetics of the purified PPD were determined, using both synthetic and natural inhibitors. Among inhibitors tested, ascorbic acid was the most effective inhibitor with the lowest Ki value of 0.36 mM. This is the first study on the purification and characterisation of PPD from purslane (Portulaca oleracea) that may provide new insight into how to overcome the enzymatic browning.Keywords: Portulaca oleracea; polyphenol oxidase; purification; kinetics; inhibitors.Practical Applications: Understanding inhibition mechanism of PPD may provide insight into plant browning in food industry.

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Purification and Partial Biochemical Characterization of Polyphenol Oxidase from Mango (Mangifera indica cv. Manila)

Cited by 34 publications

References 37 publications

Purification, partial biochemical characterization and inactivation of polyphenol oxidase from Mexican Golden Delicious apple (Malus domestica )

Purification, partial biochemical characterization and inactivation of polyphenol oxidase from Mexican Golden Delicious apple (Malus domestica )

Inhibition kinetics of oligochitosan as an uncompetitive inhibitor on fuji apple polyphenol oxidase

Purification and characterization of polyphenol oxidase from purslane

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