Characterization of a pericarp browning related LACCASE 14-4 from longan fruit with a focus on (epi)catechin oxidative polymerization

Zhu, Qiunan; Zhou, Xianfang; Zhang, Xuelian; Dang, Zhaoxia; Liang, Shijin; Li, Ganting; Zhang, Zhaoqi; Fang, Fang; Pang, Xuequn

doi:10.1016/j.postharvbio.2021.111802

Cited by 16 publications

(10 citation statements)

References 60 publications

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“…Based on PSORT program bioinformatics analyses, DkLAC1 was predicted to be localized in vacuoles, where PAs aggregate and the polymerization reaction occurs [ 12 ]. Recently, LAC14-4 was visualized in vacuoles and the endoplasmic reticulum in tobacco leaves [ 42 ]. DkLAC2 can also be detected in the vacuole of epidermal cells in a tobacco leaf ( Figure 2 B).…”

Section: Discussionmentioning

confidence: 99%

DkmiR397 Regulates Proanthocyanidin Biosynthesis via Negative Modulating DkLAC2 in Chinese PCNA Persimmon

Zaman

Zhang

Liu

et al. 2022

IJMS

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Persimmon fruits accumulate a large amount of proanthocyanidins (PAs), which makes an astringent sensation. Proanthocyanidins (PAs) are the polymers of flavan-3-ols stored in plant vacuoles under laccase activation. A laccase gene, DkLAC2, is putatively involved in PAs biosynthesis and regulated by microRNA (DkmiR397) in persimmon. However, the polymerization of PAs in association with miRNA397 still needs to be explored in persimmon. Here, we identified pre-DkmiR397 and its target gene DkLAC2 in ‘Eshi 1’ persimmon. Histochemical staining with GUS and dual luciferase assay both confirmed DkmiR397-DkLAC2 binding after co-transformation in tobacco leaves. Diverse expression patterns of DkLAC2 and DkmiR397 were exhibited during persimmon fruit development stages. Moreover, a contrasting expression pattern was also observed after the combined DkLAC2-miR397 transformation in persimmon leaves, suggesting that DkmiR397 might be a negative regulator of DkLAC2. Similarly, the transient transformation of DkmiR397 in persimmon fruit discs in vitro also reduced PA accumulation by repressing DkLAC2, whereas the up-regulation of DkLAC2 increased the accumulation of PAs by short tandem target mimic STTM-miR397. A similar expression pattern was observed when overexpressing of DkLAC2 in Arabidopsis wild type (WT) and overexpression of DkLAC2, DkmiR397 in persimmon leaf callus. Our results revealed that the role of DkmiR397 repressed the expression of DkLAC2 concerning PA biosynthesis, providing a potential target for the manipulation of PAs metabolism in persimmon.

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Section: Discussionmentioning

confidence: 99%

DkmiR397 Regulates Proanthocyanidin Biosynthesis via Negative Modulating DkLAC2 in Chinese PCNA Persimmon

Zaman

Zhang

Liu

et al. 2022

IJMS

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“…We extracted flavan-3-ols and PCs from Huaniu and Fuji peels according to the method in our previous study [ 15 ]. We first homogenized 0.1 g of freeze-dried peel with 1.5 mL of n -hexane with 1% ( w/v ) butylated hydroxytoluene (BHT) and 3% ( v/v ) formic acid to remove fat-soluble substances.…”

Section: Methodsmentioning

confidence: 99%

“…The enzyme extraction and purification were carried out using the method of Han et al [ 19 ] and our previous study [ 15 ]. We ground 200 g of frozen Huaniu peel to powder by liquid N 2 and added it to 600 mL extraction buffer (0.05 M pH 7.0 phosphate buffer (PBS), containing 50% ( w/v ) polyvinyl-polypyrrolidon (PVPP) and 1% ( w / v ) protease-inhibitor cocktail (Sigma-Aldrich, Merck, NJ, USA)).…”

Section: Methodsmentioning

confidence: 99%

“…According to our previous study [ 15 ], we performed the activity staining of the purified proteins on 10% ( w / v ) sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE), without boiling the protein sample. We rinsed the gel twice with a citrate phosphate buffer (50 mM, pH 6.0) for 5 min each time for the removal of the SDS.…”

Section: Methodsmentioning

confidence: 99%

“…For example, researchers detected oxidation polymerization products in the in vitro reaction of CT incubated with a crude PPO extract from grape [ 14 ]. Recently, we purified two laccases, LcADE/LAC and DlLAC14-4, from litchi and longan pericarp, respectively, and demonstrated their ability to catalyze the oxidative polymerization of PA monomers, such as EC and CT, and produce brown products in vitro [ 15 , 16 ]. The high levels of LcADE/LAC and DlLAC14-4 in litchi and longan pericarp, respectively, suggest that they play important roles in pericarp browning.…”

Section: Introductionmentioning

confidence: 99%

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

Zhou

Guan

Pang

et al. 2022

Foods

Self Cite

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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.

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Traditional and next-generation methods for browning control in plant tissue culture: Current insights and future directions

Permadi,

Akbari,

Prismantoro

et al. 2024

Current Plant Biology

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Characterization of a pericarp browning related LACCASE 14-4 from longan fruit with a focus on (epi)catechin oxidative polymerization

Cited by 16 publications

References 60 publications

DkmiR397 Regulates Proanthocyanidin Biosynthesis via Negative Modulating DkLAC2 in Chinese PCNA Persimmon

DkmiR397 Regulates Proanthocyanidin Biosynthesis via Negative Modulating DkLAC2 in Chinese PCNA Persimmon

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

Traditional and next-generation methods for browning control in plant tissue culture: Current insights and future directions

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