Comparison of the characterization and the temperature/pressure stability of soluble and membrane-bound polyphenol oxidase from ‘Lijiang’ snow peach

Wang, Fuhai; Zhou, Hengle; Cheng, Fengyun; Niu, Huihui; Yuan, Lei; Yi, Junjie; Zhou, Linyan

doi:10.1016/j.lwt.2021.111401

Cited by 11 publications

(4 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Researchers have also reported that 4-methylcatechol [ 29 ] and catechol [ 30 ] are the most suitable substrates for PPO purified from apples. Moreover, for the PPOs purified from round brinjal ( Solanum melongena ) [ 31 ], royal dates ( Ziziphus jujuba ) [ 32 ], lily bulb ( Lilium spp ) [ 33 ], and peach ( Prunus persica ) [ 34 ]' 4-methylcatechol is the most suitable substrate. Flavanols (mainly EC) and PCs are the most abundant phenolic substances in apples [ 8 , 10 ].…”

Section: Resultsmentioning

confidence: 99%

Purification and Characterization of a Dark Red Skin Related Dimeric Polyphenol Oxidase from Huaniu Apples

Zhou

Guan

Pang

et al. 2022

Foods

View full text Add to dashboard Cite

The distinct dark-red skin of Huaniu apples renders them attractive to customers. However, the mechanism that leads to the development of the color of the fruit is unclear. In this study, we found that compared with red Fuji (a bright-red apple cultivar), Huaniu apples had higher contents of (−)-epicatechin (EC), (−)-epigallocatechin (EGC), (−)-gallocatechin gallate (GCG), and procyanidins (PCs) B2 and C1 in the peel, which implies that the polymerization of the flavanols and PCs may be correlated with the dark-red skin of the fruit. Using EC as a substrate, we purified an enzyme from Huaniu peel. We performed protein sequencing and discovered that the enzyme was a polyphenol oxidase (PPO). The molecular weight of the enzyme was approximately 140 kDa, which we estimated by native-PAGE and SDS-PAGE, while it was 61 kDa by urea-SDS-PAGE, from which we discovered that the PPO was a dimer. We observed the lowest Km value for catechol (0.60 mM), and the best substrate was 4-methylcatechol, with a Vmax of 526.32 U mg−1 protein. EC is a suitable natural substrate, with a Km value of 1.17 mM, and 55.27% of the Vmax/Km of 4-methylcatechol. When we used EC as a substrate, the optimum temperature and pH of the PPO were 25 °C and 5.0, respectively. In summary, we purified a dimeric PPO from Huaniu apples that showed high activity to EC, which might catalyze the polymerization of flavanols and PCs and lead to the dark-red color development of the fruit.

show abstract

Section: Resultsmentioning

confidence: 99%

Purification and Characterization of a Dark Red Skin Related Dimeric Polyphenol Oxidase from Huaniu Apples

Zhou

Guan

Pang

et al. 2022

Foods

View full text Add to dashboard Cite

show abstract

“…Jia et al [6] compared the effects of inhibitors on sPPO and mPPO in peach fruit, and the results showed that L-cysteine and ascorbic acid have a stronger inhibitory effect on sPPO than mPPO. Therefore, we summarized the following points: (i) mPPO's activity was usually higher than that of sPPO; (ii) there are certain differences in the optimum substrate, pH, and temperature, it was found that mPPO was generally more resistant to acid/alkaline and pressure/temperature than sPPO; (iii) sPPO and mPPO could show different tolerance in the presence of inhibitors, and their inhibition mechanisms were also different, with mPPO typically exhibiting stronger inhibition tolerance generally [7][8][9][10][11][12]. The differences in the enzyme properties were mainly attributed to their structure, which could be determined by analyzing the amino acid sequences of the enzyme, and other information, such as the enzyme's bioinformatics, function, and evolution, could be obtained.…”

Section: Introductionmentioning

confidence: 99%

“…However, the current studies reported on the effects of HPP on the activity and conformation of PPO did not clarify the forms of PPO, limiting the further exploration of the inactivation mechanism of PPO. Wang et al [10] reported that the activities of sPPO and mPPO in 'Lijiang snow' peach after HPP at 550 MPa were decreased by 89% and 12%, respectively, indicating that mPPO exhibited more resistance to pressure than sPPO. Zhou et al [25] also reported that the residual activity of mPPO and sPPO in pear was decreased to 59.6% and 13.5% by HPP (600 MPa, 16 min), respectively.…”

Section: Introductionmentioning

confidence: 99%

Comparing the Effect of HPP on the Structure and Stability of Soluble and Membrane-Bound Polyphenol Oxidase from ‘Lijiang Snow’ Peach: Multispectroscopic and Molecular Dynamics Simulation

Zhou

Bie

et al. 2023

Foods

Self Cite

View full text Add to dashboard Cite

Polyphenol oxidase (PPO) easily causes fruits and vegetables to lose their color and nutritional value. As a non-thermal process, high-pressure processing (HPP) showed different inactivation effects on endogenous enzymes. In this work, soluble PPO (sPPO) and membrane-bound PPO (mPPO) from ‘Lijiang snow’ peaches were purified, and then the effect of high pressure on the conformation of sPPO and mPPO was investigated and compared at the molecular level. The maximum activation of sPPO and mPPO by 11.2% and 4.8% was observed after HPP at 200 MPa, while their activities both gradually decreased at 400 MPa and 600 MPa; in particular, the residual activities of sPPO and mPPO at 600 MPa for 50 min were 41.42% and 72.95%, respectively. The spectroscopic results indicated that the secondary structure of PPOs was little affected by HPP, but HPP led to obvious changes in their tertiary structure. The simulations showed that the decreasing distance between the copper ion and His residue in the copper-binding region of two PPOs at 200 MPa was favorable to catalytic activity, while the increasing distance between copper ions and His residues and the disordered movement of the loop region above 400 MPa were unfavorable. In addition, the structure of sPPO was relatively looser than that of mPPO, and high pressure showed a more significant effect on the conformation of sPPO than that of mPPO. This study clarified the effect of HPP on PPO’s structure and the relationship between its structure and activity and provided a basis for the prevention of enzymatic browning.

show abstract

“…PPO (E. C. 1.14.18.1), a type of oxidoreductase that widely exists in nature, is a copper‐containing metalloproteinase that causes browning of fruits, vegetables, fungi and crustaceans (Alici & Arabaci, 2016; Lei et al ., 2018; Wang et al ., 2021). Notably, during the processing of black tea, PPO plays an active role in oxidising tea polyphenols to o‐quinones and later in forming theaflavins (TFs) (Vermeer et al ., 2008).…”

Section: Introductionmentioning

confidence: 99%

In vitro enzymatic synthesis of a monomeric theaflavin using a polyphenol oxidase isozyme from tea (Camellia sinensis) leaf

Teng

Liu

Zeng

et al. 2021

Int J of Food Sci Tech

View full text Add to dashboard Cite

Polyphenol oxidase (PPO) plays an important role in tea production, and there are many studies on the enzymatic synthesis of theaflavins using PPO; however, using a purified PPO isozyme (PPOI) instead of a crude PPO enzyme to obtain monomeric theaflavin in vitro is a novel approach that has rarely been previously reported. In the present study, a tea PPOI was purified and used for the enzymatic synthesis of theaflavin monomers. Tea leaf PPOI was purified 12.90-fold, with recovery yield of 1.30% via the sequential use of precipitation with 30%À80% saturated (NH 4 ) 2 SO 4 , Q-Sepharose Fast Flow anion exchange chromatography and Sephadex G-75 gel filtration chromatography. Subsequently, PPOI was used in the enzymatic synthesis of theaflavins: under various pH, temperatures, reaction times and catechin substrate composition and concentration, only the amount of theaflavin components produced was changed, and no other theaflavin component apart from the theaflavin monomer was produced. These results highlight the importance of monomeric theaflavin synthesis using purified PPOI isozyme and provide insight into PPO in the context of tea production.

show abstract

Comparison of the characterization and the temperature/pressure stability of soluble and membrane-bound polyphenol oxidase from ‘Lijiang’ snow peach

Cited by 11 publications

References 42 publications

Purification and Characterization of a Dark Red Skin Related Dimeric Polyphenol Oxidase from Huaniu Apples

Purification and Characterization of a Dark Red Skin Related Dimeric Polyphenol Oxidase from Huaniu Apples

Comparing the Effect of HPP on the Structure and Stability of Soluble and Membrane-Bound Polyphenol Oxidase from ‘Lijiang Snow’ Peach: Multispectroscopic and Molecular Dynamics Simulation

In vitro enzymatic synthesis of a monomeric theaflavin using a polyphenol oxidase isozyme from tea (Camellia sinensis) leaf

Contact Info

Product

Resources

About