Skip to main content
SpringerLink
Log in

Optimization of enzymatic synthesis of theaflavins from potato polyphenol oxidase

  • Research Paper
  • Published:
Bioprocess and Biosystems Engineering Aims and scope Submit manuscript

Abstract

Theaflavin (TF), a chemical component important in measuring the quality of fermented tea, has a strong natural antioxidant effect and many pharmacological functions. Enzymatic oxidation has become a widely used method for preparing TFs at the current research stage. Using plant exogenous polyphenol oxidase (PPO) to enzymatically synthesize TFs can significantly increase yield and purity. In this study, tea polyphenols were used as the reaction substrate to discuss the optimal synthesis conditions of potato PPO enzymatic synthesis of theaflavins and the main products of enzymatic synthesis of TFs. The optimal enzymatic synthesis conditions were as follows: pH of the reaction system was 5.5, reaction time was 150 min, substrate concentration was 6.0 mg/mL, reaction temperature was 20 °C, and the maximum amount of TFs produced was 651.75 μg/mL. At the same time, high-performance liquid chromatography was used to determine the content of theaflavins and catechins in the sample to be tested, and the dynamic changes and correlations of the main catechins and theaflavins in the optimal enzymatic system were analyzed. The results showed that epicatechin (EC), epigallocatechin (EGC), epicatechin gallate (ECG), and epigallocatechin gallate (EGCG) are all the main substrates synthesis of TFs. The main substrate of TFs and its strongest enzymatic catalytic effect on EGCG make theaflavin-3,3′-digallate (TFDG) the most important synthetic monomer. In this study, theaflavins were synthesized by polyphenol oxidase catalysis, which laid a foundation for industrialization of theaflavins.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig.4
Fig. 5

Similar content being viewed by others

Abbreviations

TF:

Theaflavin

PPO:

Polyphenol oxidase

EC:

Epicatechin

EGC:

Epigallocatechin

ECG:

Epicatechin gallate

EGCG:

Epigallocatechin gallate

TFs:

Theaflavins

TFDG:

Theaflavin-3,3′-digallate

TF-3-G:

Theaflavin-3-gallate

TF-3′-G:

Theaflavin-3′-gallate

References

  1. Zhang J, Cui H, Jiang H, Xiong C (2019) Rapid determination of theaflavins by HPLC with a new monolithic column. Czech J Food Sci 37(2):112–119. https://doi.org/10.17221/213/2018-CJFS

    Article  CAS  Google Scholar 

  2. Li B, Vik SB, Tu Y (2012) Theaflavins inhibit the ATP synthase and the respiratory chain without increasing superoxide production. J Nutr Biochem. https://doi.org/10.1016/j.jnutbio.2011.05.001

    Article  PubMed  PubMed Central  Google Scholar 

  3. Jianping S, Qingyan M, Yongyuan L (2016) Theaflavins suppress tumor growth and metastasis via the blockage of the STAT3 pathway in hepatocellular carcinoma. OncoTargets Ther 9:4265–4275 (CNKI:SUN:WXQG.0.2011-01-005)

    Article  Google Scholar 

  4. de Oliveira A, Prince D, Lo C-Y, Chu T-C (2015) Antiviral activity of theaflavin digallate against herpes simplex virus type 1. Antiviral Res. https://doi.org/10.1016/j.antiviral.2015.03.009

    Article  PubMed  PubMed Central  Google Scholar 

  5. Wang HX, Sun JT, Wen-Ping LV, Chao-Yang MA, Xia WS (2011) Research progress on preparation, analysis, separation and function of theaflavins. J Food Sci Biotechnol. https://doi.org/10.2147/OTT.S102858

    Article  Google Scholar 

  6. Redfearn DP, Trim GM, Skane AC, Klein GJ (2005) Esophageal temperature monitoring during radiofrequency ablation of atrial fibrillation. J Cardiovasc Electrophysiol. https://doi.org/10.1111/j.1540-8167.2005.40825.x

    Article  PubMed  Google Scholar 

  7. Caixia L, Jingrui Y, Guangying W, He N, Haihang L (2017) Research on the synthesis of theaflavins catalyzed by polyphenol oxidase. Chin J Plant Physiol 08:1359–1364. https://doi.org/10.13592/j.cnki.ppj.2017.0095

    Article  Google Scholar 

  8. Sufang L, Shengpeng W (2015) Research on tea pigment and its biological activity function. Hubei Agric Sci 24:6117–6119. https://doi.org/10.14088/j.cnki.issn0439-8114.2015.24.002

    Article  Google Scholar 

  9. Huang Yingjie Wu, Mengyao YY, Youyi H (2017) The effect of different reaction conditions on the synthesis of theaflavins by polyphenol oxidase from Mengku large leaf species. Food Sci 22:54–59

    Google Scholar 

  10. Matsuo Y, Tadakuma F, Shii T, Tanaka T (2015) Selective oxidation of pyrogallol-type catechins with unripe fruit homogenate of Citrus unshiu and structural revision of oolongtheanins. Tetrahedron. https://doi.org/10.1016/j.tet.2015.03.016

    Article  Google Scholar 

  11. van der Westhuizen M, Steenkamp L, Steenkamp P, Apostolides Z (2015) Alternative pathway implicated as an influencing factor in the synthesis of theaflavin. Biocatal Biotransform. https://doi.org/10.3109/10242422.2016.1163341

    Article  Google Scholar 

  12. Lei S, Xie M, Hu B, Zeng X (2017) Effective synthesis of theaflavin-3,3′-digallate with epigallocatechin-3-O-gallate and epicatechin gallate as substrates by using immobilized pear polyphenol oxidase. Int J Biol Macromol. https://doi.org/10.1016/j.ijbiomac.2016.10.072

    Article  PubMed  Google Scholar 

  13. Shenghu C (2016) Isolation and purification of polyphenol oxidase isoenzymes from Longjing No. 43 tea tree and the properties of PPO III-2 and PPO V-3. Huazhong Agricultural University. http://gffiy28995338bdc041dasxx5k9pnf5xvn6xqw.fffb.suse.cwkeji.cn:999/KCMS/detail/detail.aspx?dbname=CMFD201701&filename=1016155372.nh. Accessed 7 Apr 2022

  14. Lei X (2014) Extraction, isolation and purification of tea tree polyphenol oxidase and its partial enzyme properties. Huazhong Agricultural University. http://gffiy28995338bdc041dasxx5k9pnf5xvn6xqw.fffb.suse.cwkeji.cn:999/KCMS/detail/detail.aspx?dbname=CMFD201402&filename=1014213740.nh. Accessed 7 Apr 2022

  15. Teng J et al (2017) Purification, characterization and enzymatic synthesis of theaflavins of polyphenol oxidase isozymes from tea leaf (Camellia sinensis). LWT 84:263–270. https://doi.org/10.1016/j.lwt.2017.05.065

    Article  CAS  Google Scholar 

  16. Health U D O, Services H (2001) Guidance for industry, bioanalytical method validation [J]. http://www.fda.gov/cder/guidance/index.htm. Accessed 8 Apr 2022

  17. Sitheeque MAM, Panagoda GJ, Yau J, Samaranayake LP (2009) Antifungal activity of black tea polyphenols (catechins and theaflavins) against Candida species. Chemotherapy 55(3):189–196

    Article  CAS  Google Scholar 

  18. Subramanian N, Venkatesh P, Ganguli S, Sinkar VP (1999) Role of polyphenol oxidase and peroxidase in the generation of black tea theaflavins. J Agric Food Chem 47(7):2571

  19. Alastair R (1983) Effects of physical and chemical conditions on the in vitro oxidation of tea leaf catechins. Phytochem 22(4):889–896

    Article  Google Scholar 

  20. Yiyi C, Guanghui J (1993) Theoretical discussion on temperature-variable fermentation of black tea. Tea Sci 02:81–86

    Google Scholar 

  21. Tanmoy S, Vijayakumar C, Shrilekha D, Adinpunya M (2015) Assessing biochemical changes during standardization of fermentation time and temperature for manufacturing quality black tea. J Food Sci Technol 52(4):2387–2393

    Article  Google Scholar 

  22. Weixiang X, Chun Li, Hui X (1992) A preliminary study on the formation and degradation of black tea pigments. Tea Sci 01:49–54

    Google Scholar 

  23. Jing Fu, Heyuan J, Jianyong Z, Liting S, Weiwei W (2019) Research progress in the synthesis of catechin dimer oxidation products catalyzed by exogenous polyphenol oxidase. Food Sci 07:274–280

    Google Scholar 

  24. Davies AP, Goodsall C, Cai Y, Davis AL, Nurstend HE (1999) Black tea dimeric and oligomeric pigments—structures and formation. Springer US, Boston

    Book  Google Scholar 

  25. Takashi T, Chie M, Kyoko I, Isao K (2002) Synthesis of theaflavin from epicatechin and epigallocatechin by plant homogenates and role of epicatechin quinone in the synthesis and degradation of theaflavin. J Agric Food Chem 50(7):2142–2148

    Article  Google Scholar 

  26. Dai XL (2020) Discovery and characterization of tannase genes in plants: roles in hydrolysis of tannins. New Phytol 226(4):1104–1116

    Article  CAS  Google Scholar 

  27. Coggon P, Moss GA, Graham HN, Sanderson GW (1973) The biochemistry of tea fermentation: oxidative degallation and epimerization of the tea flavanol gallates. J Agric Food Chem 21(4):727–733

  28. Bajaj KL, Anan T, Tsushida T, Ikegaya K (2006) Effects of (−)-epicatechin on oxidation of theaflavins by polyphenol oxidase from tea leaves. J Agric Chem Soci Jpn 51(7):1767–1772

    Google Scholar 

Download references

Acknowledgements

This work was financially supported by the Project of Enzymatic preparation of theaflavin from plant exogenous polyphenol oxidase of China (Grant no. 2021084), the Construction and demonstration of modern selenium rich black tea industry in Pingshan, China (Grant no. 2017NFP1068) and the Construction and application of modern Matcha production base in Pingshan, China (Grant no. 2018NZYZF0116).

Author information

Author notes

  1. Dong Li and Liang Dong contributed equally to this work.

Authors and Affiliations

  1. College of Bioengineering, Sichuan University of Science and Engineering, Yibin, 644000, Sichuan, People’s Republic of China

    Dong Li, Liang Dong, Jieyuan Li, Shiqi Zhang, Yu Lei, Mengsheng Deng & Jingya Li

Authors
  1. Dong Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Liang Dong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jieyuan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shiqi Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yu Lei
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mengsheng Deng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jingya Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shiqi Zhang.

Ethics declarations

Conflict of interest

The authors have no conflicts of interest to report.

Ethical statement

I declare that I abide by the academic ethics, advocating rigorous style of study. This article is the authors’ study results. No competitive interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, D., Dong, L., Li, J. et al. Optimization of enzymatic synthesis of theaflavins from potato polyphenol oxidase. Bioprocess Biosyst Eng 45, 1047–1055 (2022). https://doi.org/10.1007/s00449-022-02723-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00449-022-02723-x

Keywords

Search

Navigation