Purification of Polyphenol Oxidase from Tea Leaf by Three Phase Partitioning and Enzymatic Properties

LIU Yang; CHEN Qincao; LIU Dechun; YANG Li; HU Wei; KUANG Liuqing; LIU Yong; TENG Jie

doi:10.13386/j.issn1002-0306.2021080075

Volume 43 Issue 9

Apr. 2022

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Science and Technology of Food Industry > 2022 > 43(9): 119-124. > DOI: 10.13386/j.issn1002-0306.2021080075

LIU Yang, CHEN Qincao, LIU Dechun, et al. Purification of Polyphenol Oxidase from Tea Leaf by Three Phase Partitioning and Enzymatic Properties[J]. Science and Technology of Food Industry, 2022, 43(9): 119−124. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021080075.

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Purification of Polyphenol Oxidase from Tea Leaf by Three Phase Partitioning and Enzymatic Properties

College of Agronomy, Jiangxi Agricultural University, Nanchang 330045, China

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Received Date: August 05, 2021
Available Online: February 27, 2022

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Abstract

In order to study a fast and efficient method for obtain polyphenol oxidase (PPO) from tea leaf, three phase partitioning (TPP) were used to purified PPO and then its enzymatic properties were studied. The results showed that PPO from tea leaf was purified with 15.76-fold enrichment in the specific activity of 210.46 U/mg and 8.04% recovery by TPP purification method for the second time. The strongest affinity substrate was pyrogallol (K_m=6.82 mmol/L, V_max=5.36×10⁻² OD/min) for purified tea PPO. The optimum temperature and pH of the enzyme were 45 ℃ and 5.5, respectively. The enzyme had great stability under condition of 25 ℃ and pH4.5~7.0. The most significant inhibitory effect of different compounds on purified tea PPO was ascorbic acid (IC₅₀=2.42±0.18 mmol/L), followed by oxalic acid and EDTA. The inhibitory effect of citric acid was the weakest, while halides had no significant inhibitory effect (P>0.05). This work proves that the TPP method can better retain the tea PPO activity, and easy to operate, time-consuming and low-cost for the purification of tea PPO, which is beneficial to industrial applications.
- tea leaf,
- three phase partitioning (TPP),
- polyphenol oxidase (PPO),
- enzymatic properties

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Science and Technology of Food Industry

Purification of Polyphenol Oxidase from Tea Leaf by Three Phase Partitioning and Enzymatic Properties

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