Preparation of a Cross-linked Polyphenol Oxidase Aggregate Based on Catechins and Its Efficient Catalytic Synthesis of Theaflavin-3,3&#x002b9;-digalate

ZHOU Jinghui; LIU Changwei; ZHANG Sheng; XU Gang; XU Wei; HUANG Jian&#x02032;an; LIU Zhonghua

doi:10.13305/j.cnki.jts.2023.03.005

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Journal of Tea Science ›› 2023, Vol. 43 ›› Issue (3) : 377-388. DOI: 10.13305/j.cnki.jts.2023.03.005

Preparation of a Cross-linked Polyphenol Oxidase Aggregate Based on Catechins and Its Efficient Catalytic Synthesis of Theaflavin-3,3ʹ-digalate

ZHOU Jinghui^1,2,3,4,5, LIU Changwei^1,2,3,4, ZHANG Sheng^1,2,3,4, XU Gang⁵, XU Wei⁶, HUANG Jian′an^1,2,3,4, LIU Zhonghua^1,2,3,4,*

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Abstract

In order to prepare theaflavin more efficiently and economically, catechins were used as the cross-linking reagent in this study, and a polyphenol oxidase (tyrosinase) crossed-linked aggregate derived from Bacillus megaterium was prepared and used for the synthesis of theaflavin-3,3ʹ-digalate. Through optimization of enzyme cross-linking parameters and the catalytic performance before and after cross-linking, the results show that the optimal enzyme activity recovery rate of cross-linked enzyme (200 U·mL^-1) could be obtained at pH 4.0, EGCG 0.5 mg·mL^-1 and cross-linking time 50 min. Compared with free enzymes, cross-linked enzymes showed better catalytic performance (thermal stability, organic solvent tolerance, substrate tolerance). When the cross-linked enzyme was used to synthesize theaflavin-3,3′-digallate, the concentration of the product could reach 800 μg·mL^-1, and the cross-linked enzyme could be reused for at least three batches. This method could significantly reduce the application cost of theaflavin, which has potential industrial application value.

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enzyme cross-linked aggregates / preparation / theaflavins / tyrosinase

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ZHOU Jinghui, LIU Changwei, ZHANG Sheng, XU Gang, XU Wei, HUANG Jian′an, LIU Zhonghua. Preparation of a Cross-linked Polyphenol Oxidase Aggregate Based on Catechins and Its Efficient Catalytic Synthesis of Theaflavin-3,3ʹ-digalate[J]. Journal of Tea Science. 2023, 43(3): 377-388 https://doi.org/10.13305/j.cnki.jts.2023.03.005

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