Polyphenol Oxidase from Dendrobium officinale Flowers: Characterization and Browning-Promoting Mechanism

IF 6.2 1区农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY

Journal of Agricultural and Food Chemistry Pub Date : 2025-10-11 DOI:10.1021/acs.jafc.5c03738

Xiwen Zhong, Huasi Lin, Bing Zheng, Qiang Yu, Jiayan Xie, Qi Shu, Xiaobo Hu

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Abstract

Polyphenol oxidase (PPO) triggered browning during the storage and processing of Dendrobium officinale flower (DoF), impairing their nutritional and aesthetic qualities. This study aimed to investigate the enzymatic properties and enzymatic browning mechanisms of PPO in DoF, providing a theoretical foundation for developing effective strategies to inhibit enzymatic browning. PPO with high purity was obtained by ammonium sulfate precipitation and ion-exchange chromatography, further identified by electrophoresis and LC-MS/MS. The PPO exhibited optimal activity at 35 °C and pH 6.5, while ascorbic acid, glutathione, and L-cysteine significantly inhibited its activity and furanones demonstrated potential inhibitory effects. Moreover, caffeic acid, identified as the most active PPO substrate in DoF, was determined to be the primary contributor to browning. Molecular docking revealed that caffeic acid exhibited stronger interactions with PPO compared with other phenolic compounds, which further revealed the substrate selection mechanism of PPO and the molecular basis of DoF browning.

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铁皮石斛花多酚氧化酶：表征及促褐化机制

多酚氧化酶（PPO）在铁皮石斛花（DoF）贮藏和加工过程中引发褐变，影响其营养和美学品质。本研究旨在探讨多酚氧化酶在DoF中的酶促褐变特性和酶促褐变机制，为制定有效的酶促褐变抑制策略提供理论基础。通过硫酸铵沉淀法和离子交换色谱法获得了纯度较高的PPO，并通过电泳和LC-MS/MS进行了鉴定。PPO在35°C和pH 6.5条件下表现出最佳活性，抗坏血酸、谷胱甘肽和l -半胱氨酸显著抑制其活性，呋喃酮类具有潜在的抑制作用。此外，咖啡酸被确定为DoF中最活跃的PPO底物，被确定为褐变的主要贡献者。分子对接发现，相较于其他酚类化合物，咖啡酸与PPO的相互作用更强，进一步揭示了PPO的底物选择机制和DoF褐变的分子基础。

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来源期刊

Journal of Agricultural and Food Chemistry 农林科学-农业综合

CiteScore

9.90

自引率

8.20%

发文量

1375

审稿时长

2.3 months

期刊介绍： The Journal of Agricultural and Food Chemistry publishes high-quality, cutting edge original research representing complete studies and research advances dealing with the chemistry and biochemistry of agriculture and food. The Journal also encourages papers with chemistry and/or biochemistry as a major component combined with biological/sensory/nutritional/toxicological evaluation related to agriculture and/or food.