Hysteresis and Positive Cooperativity of Iceberg Lettuce Polyphenol Oxidase

doi:10.1006/bbrc.2001.6014

Biochemical and Biophysical Research Communications

Volume 289, Issue 3, 7 December 2001, Pages 769-775

https://doi.org/10.1006/bbrc.2001.6014 Get rights and content

Abstract

A kinetic study of the diphenolase activity of latent polyphenol oxidase (PPO), purified from Iceberg lettuce (Lactuca sativa L), revealed a sigmoid relationship between the reaction rate and the substrate concentration with a high Hill coefficient (n_H = 3.8). This positive cooperativity had not been previously described for any PPO. Furthermore, the enzyme showed a lag phase in the expression of this activity, suggesting a hysteretic nature of the enzyme. The kinetic behavior, the latency and the lag phase varied at different steps of the purification process. PPO showed hyperbolic or cooperative kinetics depending on the pH assay and the sodium dodecyl sulfate (SDS) concentration. Substrate-induced slow conformational change of the oligomeric enzyme is suggested. The conformational change would be toward a more active enzyme form with higher affinity for the substrate and favoured by acid pH and SDS.

References (39)

P. Thipyapong et al.
Systemic wound induction of potato (Solanum tuberosum) polyphenol oxidase
Phytochemistry
(1995)
J. Cabanes et al.
A kinetic study of the melanization pathway between $l$ -tyrosine and dopachrome
Biochim. Biophys. Acta
(1987)
J.N. Rodrı́guez-López et al.
Analysis of a kinetic model for melanin biosynthesis pathway
J. Biol. Chem.
(1992)
A. Sanchez-Ferrer et al.
Tyrosinase: A comprehensive review of its mechanism
Biochim. Biophys. Acta
(1995)
M. Jiménez et al.
The effect of sodium dodecyl sulphate on polyphenol oxidase
Phytochemistry
(1996)
T. Swain et al.
Activation of Vicia faba (L.) tyrosinase as effected by denaturing agents
Phytochemistry
(1966)
C. Wittenberg et al.
A detergent-activated tyrosinase from Xenopus laevis. II Detergent activation and binding
J. Biol. Chem.
(1985)
C.W.G. van Gelder et al.
Sequence and structural features of plant and fungal tyrosinases
Phytochemistry
(1997)
H.R. Lerner et al.
Evidence for conformational changes in grape catechol oxidase
Phytochemistry
(1972)
H.R. Lerner et al.
Stokes radius changes of solubilized grape catechol oxidase
Phytochemistry
(1975)

E. Núñez-Delicado et al.

Triton X-114 aided purification of latent tyrosinase

J. Chromatogr. B.

(1996)

J. Monod et al.

On the nature of allosteric transitions: A plausible model

J. Mol. Biol.

(1965)

B.I. Kurganov et al.

The theoretical analysis of kinetic behaviour of “histeretic” allosteric enzymes. I. The kinetic manifestations of slow conformational change of an oligomeric enzyme in the Monod, Wyman, and Changeux model

J. Theor. Biol.

(1976)

M.M. Bradford

A rapid and sensitive method for the quantification of microgram quantities of proteins utilizing the principle of protein-dye binding

Anal. Biochem.

(1976)

M. Jiménez et al.

Kinetics of the slow pH-mediated transition of polyphenol oxidase

Arch. Biochem. Biophys.

(1996)

J.H. Waite

Calculating extinction coefficients for enzymatically produced o-quinones

Anal. Biochem.

(1976)

C. Frieden

Kinetic aspects of regulation of metabolic processes

J. Biol. Chem.

(1970)

J. Pawelek et al.

The enzymology of melanogenesis

G. Prota et al.

The chemistry of Melanism and related metabolites

Cited by (36)

A comprehensive metabolomics analysis of Torreya grandis nuts with the effective de-astringent treatment during the postharvest ripening stage
2023, Food Chemistry
Citation Excerpt :
The CIRG value change was consistent with the color change (Fig. 1B). It has been reported that the color change in the seed coat may be caused by a change in the phenolic substances after oxidization (Chazarra, Garcia-Carmona, & Cabanes, 2001). Indeed, the total phenol content in the seed coat of the two de-astringent treatments displayed a decreasing trend, and the nuts treated with NaHCO3 showed a more pronounced decreasing trend compared to those under the CK treatment, from 56.47 mg g−1 on Day 0 to 2.63 mg g−1 on Day 10 (Fig. 1C).
Astringency removal is important for the quality of Torreya grandis nut and occurs after harvest. Here, we evaluated the effect of NaHCO₃ treatment on astringency removal and compared the differential metabolites of the seed coat and kernel using a UHPLC QQQ-MS-based metabolomics approach. The result revealed the nut astringency was primarily enriched in the seed coat with more soluble tannins. The NaHCO₃ treatment greatly shortened the de-astringency process, as indicated by a faster conversion of soluble tannins to insoluble tannins and more acetaldehyde production. Besides, a total of 293 metabolites, including 92 phenolic acids and 37 flavonoids, were tentatively characterized in the seed coat. A further comparative analysis of the metabolomics indicated epigallocatechin, gallocatechin, catechin, procyanidin B1, B2, B3 and C1 to be the major metabolites influenced by the NaHCO₃ treatment. This study provides new insights regarding the metabolite differences of Torreya grandis nuts processed with different de-astringent treatments.
High-pressure carbon dioxide treatment and vacuum packaging alleviate the yellowing of peeled Chinese water chestnut (Eleocharis tuberosa)
2022, Food Packaging and Shelf Life
Citation Excerpt :
This might be due to the reason that peeled CWC in control group were exposed to oxygen during storage which caused the accumulation of reactive oxygen species (ROS), consequently damaging the cell membrane structure and loss of regionalization function (Wang et al., 2022). The increase in PPO activity could be due to the contact between PPO and substrate, which led to the allosterism of PPO towards the higher catalytic activity structure (Chazarra et al., 2001). In addition to polyphenolic substrates and PPO, the occurrence of enzymatic browning requires the participation of oxygen (Murtaza et al., 2018).
This work evaluated the effects of high-pressure carbon dioxide (HPCD) treatment and vacuum packaging on the yellowing of peeled Chinese water chestnut (CWC) during storage. The changes in color, enzymatic activities, total phenolic content and malondialdehyde contents were assessed in peeled CWC. The results showed that vacuum packaging alone, atmospheric packaging after HPCD treatment, and vacuum packaging after HPCD treatment (HPCD+VP) could significantly delay the yellowing of fresh-cut CWC to varying degrees. The total phenolic content was lower (68%) in HPCD+VP samples than that of control. Compared to control, the enzymatic activities such as phenylalanine ammonia-lyase, polyphenol oxidase and lipoxygenase decreased by 76% (12th day), 24% (6th day) and 19% (6th day) respectively in HPCD+VP samples. HPCD and vacuum packaging could efficiently inhibit several enzymes activities. Our study exhibited that HPCD+VP treatment delayed the yellowing of fresh-cut CWC by reducing the phenylpropanoid metabolism pathway activity and cell membrane peroxidation.
Two-year comparison of the biochemical properties of polyphenol oxidase from Turkish Alyanak apricot (Prunus armenica L.)
2016, Food Chemistry
Citation Excerpt :
Interaction of o-quinones with other molecules leads to formation of red, brown, or black pigments, which lower quality attributes and economic values of raw material and products. Enzymatic browning is regarded as a serious problem that can occur during harvesting, handling, processing, and storage of fruits and vegetables (Chararra, Carcia-Carmona, & Cabanes, 2001; Gauillard & Richard-Forget, 1997; Ramirez, Whitaker, & Virador, 2003; Zawistowski, Biliaderis, & Eskin, 1991). There are many reports related to purification and characterization of PPO from various fruits and vegetables, e.g. mamey (Palma-Orozco, Ortiz-Moreno, Dorantes-Álvarez, Sampedro, & Nájera, 2011), eggplant (Mishra, Gautam, & Sharma, 2012), wheat (Altunkaya & Gökmen, 2012), mushroom (Kolcuoğlu, 2012), snake fruit (Zaini, Osman, Hamid, Ebrahimpour, & Saari, 2013), Lonicera japonica (Liu et al., 2013), persimmon (Navarro, Tárrega, Miguel, & Enrique, 2014), Solanum lycocarpum (Batista, Batista, Alves, & Fernandes, 2014).
Polyphenol oxidase (PPO) was extracted and purified from Turkish Alyanak apricot variety and its characteristics were studied in two consecutive years (2008 and 2009). Three isoenzymes (isoenzyme A1, A2 and B) were obtained upon ammonium sulfate fractionation, ion exchange chromatography using DEAE-Toyopearl 650 M and gel filtration chromatography using Sephadex G-100. The isoenzymes exhibited different kinetic properties. Furthermore, year-to-year variability in K_m and V_max values was significant. The pH optimum for enzyme activity was 4.98 for isoenzymes A1 and A2, and 5.8 for isoenzyme B. The isoenzymes A1 and B had optimum temperature at 30 °C in both years whereas isoenzyme A2 had maximum activity at 40 °C in 2008 and 30 °C in 2009. The inactivation kinetics parameters and the effects of inhibitors tested exhibited significant year-to-year variation.
Blueberry polyphenol oxidase: Characterization and the kinetics of thermal and high pressure activation and inactivation
2015, Food Chemistry
Citation Excerpt :
There are a number of reports in literature where an increase in the activity of plant PPOs was observed following heat treatment (Buckow, Weiss, & Knorr, 2009; Yemenicioglu, Ozkan, & Cemeroglu, 1997), which has been attributed to the activation of the latent PPO precursor in the PPO extracts. The presence of latent PPOs has been reported in many fruits and vegetables including, beetroot (Gandia-Herrero, Jimenez-Atienzar, Cabanes, Garcia-Carmona, & Escribano, 2005), iceberg lettuce (Chazarra, Garcia-Carmona, & Cabanes, 2001), apples (Yemenicioglu et al., 1997), grapes (Nunez-Delicado, Serrano-Megias, Perez-Lopez, & Lopez-Nicolas, 2007), persimmon (Nunez-Delicado, Sojo, Garcia-Carmona, & Sanchez-Ferrer, 2003) and peach (Winters, Minchin, Michaelson-Yeates, Lee, & Morris, 2008). Gerdemann, Eicken, Gala, and Krebs (2002) modelled the structure of the latent PPO from Sweet potatoes based on the 3D structural data of the active (mature) sweet potato PPO and hemocyanin from giant octopus, which lacks diphenolase activity.
Partially purified blueberry polyphenol oxidase (PPO) in Mcllvaine buffer (pH = 3.6, typical pH of blueberry juice) was subjected to processing at isothermal–isobaric conditions at temperatures from 30 to 80 °C and pressure from 0.1 to 700 MPa. High pressure processing at 30–50 °C at all pressures studied caused irreversible PPO activity increase with a maximum of 6.1 fold increase at 500 MPa and 30 °C. Treatments at mild pressure-mild temperature conditions (0.1–400 MPa, 60 °C) also caused up to 3 fold PPO activity increase. Initial activity increase followed by a decrease occurred at relatively high pressure-mild temperature (400–600 MPa, 60 °C) and mild pressure-high temperature (0.1–400 MPa, 70–80 °C) combinations. At temperatures higher than 76 °C, monotonic decrease in PPO activity occurred at 0.1 MPa and pressures higher than 500 MPa. The activation/inactivation kinetics of the enzyme was successfully modelled assuming consecutive reactions in series with activation followed by inactivation.
Dandelion PPO-1/PPO-2 domain-swaps: The C-terminal domain modulates the pH optimum and the linker affects SDS-mediated activation and stability
2015, Biochimica et Biophysica Acta - Proteins and Proteomics
Citation Excerpt :
This has led to the assumption that only the N-terminal domain specifies the catalytic properties of the enzyme. However, because PPOs can be activated in vitro by non-proteolytic reagents such as SDS [12,21–25], it is conceivable that both the linker and the C-terminal domain may be involved in this activation process and may even contribute to the enzymatic characteristics of PPOs. This is supported by the observation that the coordinated movement of the hemocyanin (HC) blocking domain away from the active site is triggered by SDS, although the overall structure of the protein is not affected.
Plant polyphenol oxidases (PPOs) have a conserved three-domain structure: (i) the N-terminal domain (containing the active site) is connected via (ii) a linker to (iii) the C-terminal domain. The latter covers the active site, thereby maintaining the enzyme in a latent state. Activation can be achieved with SDS but little is known about the mechanism. We prepared domain-swap variants of dandelion PPO-1 and PPO-2 to test the specific functions of individual domains and their impact on enzyme characteristics.
Our experiments revealed that the C-terminal domain modulates the pH optimum curve and has a strong influence on the optimal pH value. The linker determines the SDS concentration required for full activation. It also influences the SDS concentration required for half maximal activation (k_SDS) and the stability of the enzyme during prolonged incubation in buffers containing SDS, but the N-terminal domain has the strongest effect on these parameters. The N-terminal domain also determines the IC₅₀ of SDS and the stability in buffers containing or lacking SDS.
We propose that the linker and C-terminal domain fine-tune the activation of plant PPOs. The C-terminal domain adjusts the pH optimum and the linker probably contains an SDS-binding/interaction site that influences inactivation and determines the SDS concentration required for activation. For the first time, we have determined the influence of the three PPO domains on enzyme activation and stability providing insight into the regulation and activation mechanisms of type-3 copper proteins in general.
Hysteresis and positive cooperativity as possible regulatory mechanisms of Trypanosoma cruzi hexokinase activity
2014, Molecular and Biochemical Parasitology
In Trypanosoma cruzi, the causal agent of Chagas disease, the first six or seven steps of glycolysis are compartmentalized in glycosomes, which are authentic but specialized peroxisomes. Hexokinase (HK), the first enzyme in the glycolytic pathway, has been an important research object, particularly as a potential drug target. Here we present the results of a specific kinetics study of the native HK from T. cruzi epimastigotes; a sigmoidal behavior was apparent when the velocity of the reaction was determined as a function of the concentration of its substrates, glucose and ATP. This behavior was only observed at low enzyme concentration, while at high concentration classical Michaelis–Menten kinetics was displayed. The progress curve of the enzyme's activity displays a lag phase of which the length is dependent on the protein concentration, suggesting that HK is a hysteretic enzyme. The hysteretic behavior may be attributed to slow changes in the conformation of T. cruzi HK as a response to variations of glucose and ATP concentrations in the glycosomal matrix. Variations in HK's substrate concentrations within the glycosomes may be due to variations in the trypanosome's environment. The hysteretic and cooperative behavior of the enzyme may be a form of regulation by which the parasite can more readily adapt to these environmental changes, occurring within each of its hosts, or during the early phase of transition to a new host.

View all citing articles on Scopus

^☆: Abbreviations used: K_H, Hill constant; n_H, Hill coefficient; PPO, polyphenol oxidase; 4tBC, 4-tert-butylcatechol; τ, lag period; SDS, sodium dodecyl sulfate.

¹: To whom correspondence should be addressed at Departamento de Bioquı́mica y Biologı́a Molecular A, Edificio de Veterinaria, Unidad docente de Biologı́a, Universidad de Murcia, Campus de Espinardo 30071 Murcia, Spain. Fax: 968-364147. E-mail: [email protected].

View full text

Regular ArticleHysteresis and Positive Cooperativity of Iceberg Lettuce Polyphenol Oxidase☆

Abstract

Phytochemistry

Biochim. Biophys. Acta

J. Biol. Chem.

Biochim. Biophys. Acta

Phytochemistry

Phytochemistry

J. Biol. Chem.

Phytochemistry

Phytochemistry

Phytochemistry

J. Chromatogr. B.

J. Mol. Biol.

J. Theor. Biol.

Anal. Biochem.

Arch. Biochem. Biophys.

Anal. Biochem.

J. Biol. Chem.

The enzymology of melanogenesis

The chemistry of Melanism and related metabolites

Regular Article
Hysteresis and Positive Cooperativity of Iceberg Lettuce Polyphenol Oxidase☆