Abstract
Hydroxytyrosol (HT) is a diphenolic compound prevalent mainly in olives with pronounced antioxidant activity and proven benefits for human health. Current production limitations have motivated studies concerning the hydroxylation of tyrosol to HT with tyrosinase; however, accumulation of the diphenol is restricted due to its rapid subsequent oxidation to 3,4-quinone-phenylethanol. In this study, a continuous two-enzyme reaction system of sol-gel-immobilized tyrosinase and glucose dehydrogenase (GDH) was developed for the synthesis of HT. Purified tyrosinase from Bacillus megaterium (TyrBm) and E. coli cell extract expressing GDH from B. megaterium were encapsulated in a sol-gel matrix based on triethoxysilane precursors. While tyrosinase oxidized tyrosol to 3,4-quinone-phenylethanol, GDH catalyzed the simultaneous reduction of the cofactor NAD+ to NADH, which was the reducing agent enabling the accumulation of HT. Using 50 mM tyrosol, the immobilized system under optimized conditions, enabled a final HT yield of 7.68 g/L with productivity of 2.30 mg HT/mg TyrBm beads. Furthermore, the immobilized bi-enzyme system showed the feasibility for HT production from 1 mM tyrosol using a 0.5-L bioreactor as well as stable activity over 8 repeated cycles. The production of other diphenols with commercial importance such as l-dopa (3,4-dihydroxyphenylalanine) or piceatannol may be synthesized with this efficient approach.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Achmon Y, Fishman A (2015) The antioxidant hydroxytyrosol: biotechnological production challenges and opportunities. Appl Microbiol Biotechnol 99(3):1119–1130
Allouche N, Sayadi S (2005) Synthesis of hydroxytyrosol, 2-hydroxyphenylacetic acid, and 3-hydroxyphenylacetic acid by differential conversion of tyrosol isomers using Serratia marcescens strain. J Agric Food Chem 53(16):6525–6530
Allouche N, Damak M, Ellouz R, Sayadi S (2004) Use of whole cells of Pseudomonas aeruginosa for synthesis of the antioxidant hydroxytyrosol via conversion of tyrosol. Appl Environ Microbiol 70(4):2105–2109
Ates S, Cortenlioglu E, Bayraktar E, Mehmetoglu U (2007) Production of L-DOPA using Cu-alginate gel immobilized tyrosinase in a batch and packed bed reactor. Enzyme Microb Technol 40(4):683–687
Avnir D, Braun S, Lev O, Ottolenghi M (1994) Enzymes and other proteins entrapped in sol-gel materials. Chem Mater 6(10):1605–1614
Bernini R, Merendino N, Romani A, Velotti F (2013) Naturally occurring hydroxytyrosol: synthesis and anticancer potential. Curr Med Chem 20(5):655–670
Bouallagui Z, Sayadi S (2006) Production of high hydroxytyrosol yields via tyrosol conversion by Pseudomonas aeruginosa immobilized resting cells. J Agric Food Chem 54(26):9906–9911
Bouallagui Z, Sayadi S (2018) Bioconversion of p-tyrosol into hydroxytyrosol under bench-scale fermentation. Biomed Res Int 2018(7390751):1–5
Britton J, Davis R, O’Connor KE (2019) Chemical, physical and biotechnological approaches to the production of the potent antioxidant hydroxytyrosol. Appl Microbiol Biotechnol 103(15):5957–5974
Brooks SJ, Doyle EM, O’Connor KE (2006) Tyrosol to hydroxytyrosol biotransformation by immobilised cell extracts of Pseudomonas putida F6. Enzyme Microb Technol 39(2):191–196
Brouk M, Fishman A (2012) Improving process conditions of hydroxytyrosol synthesis by toluene-4-monooxygenase. J Mol Catal B Enzym 84:121–127
Choo HJ, Kim EJ, Kim SY, Lee Y, Kim B-G, Ahn J-H (2018) Microbial synthesis of hydroxytyrosol and hydroxysalidroside. Appl Biol Chem 61(3):295–301
Chung D, Kim SY, Ahn J-H (2017) Production of three phenylethanoids, tyrosol, hydroxytyrosol, and salidroside, using plant genes expressing in Escherichia coli. Sci Rep 7(1):2578
Cieńska M, Labus K, Lewańczuk M, Koźlecki T, Liesiene J, Bryjak J (2016) Effective L-tyrosine hydroxylation by native and immobilized tyrosinase. PLoS One 11(10):e0164213
Claus H, Decker H (2006) Bacterial tyrosinases. Syst Appl Microbiol 29(1):3–14
Durán N, Rosa MA, D’Annibale A, Gianfreda L (2002) Applications of laccases and tyrosinases (phenoloxidases) immobilized on different supports: a review. Enzyme Microb Technol 31(7):907–931
EFSA (2011) EFSA panel on dietetic products, nutrition and allergies. Scientific opinion on the substantiation of health claims related to polyphenols in olive. EFSA J 9(4):2033–2058
Espín JC, Soler-Rivas C, Cantos E, Tomás-Barberán FA, Wichers HJ (2001) Synthesis of the antioxidant hydroxytyrosol using tyrosinase as biocatalyst. J Agric Food Chem 49(3):1187–1193
Faccio G, Kruus K, Saloheimo M, Thöny-Meyer L (2012) Bacterial tyrosinases and their applications. Process Biochem 47(12):1749–1760
Gihaz S, Weiser D, Dror A, Sátorhelyi P, Jerabek-Willemsen M, Poppe L, Fishman A (2016) Creating an efficient methanol-stable biocatalyst by protein and immobilization engineering steps towards efficient biosynthesis of biodiesel. ChemSusChem 9(22):3161–3170
Goldfeder M, Egozy M, Shuster Ben-Yosef V, Adir N, Fishman A (2013) Changes in tyrosinase specificity by ionic liquids and sodium dodecyl sulfate. Appl Microbiol Biotechnol 97(5):1953–1961
Halaouli S, Asther M, Kruus K, Guo L, Hamdi M, Sigoillot JC, Asther M, Lomascolo A (2005) Characterization of a new tyrosinase from Pycnoporus species with high potential for food technological applications. J Appl Microbiol 98(2):332–343
Halaouli S, Asther M, Sigoillot JC, Hamdi M, Lomascolo A (2006) Fungal tyrosinases: new prospects in molecular characteristics, bioengineering and biotechnological applications. J Appl Microbiol 100(2):219–232
Hanefeld U, Gardossi L, Magner E (2009) Understanding enzyme immobilisation. Chem Soc Rev 38(2):453–468
Horvat M, Fritsche S, Kourist R, Winkler M (2019) Characterization of type IV carboxylate reductases (CARs) for whole cell-mediated preparation of 3-hydroxytyrosol. ChemCatChem 2019(11):1–12
Hoyoux A, Blaise V, Collins T, D’Amico S, Gratia E, Huston AL, Marx J-C, Sonan G, Zeng Y, Feller G (2004) Extreme catalysts from low-temperature environments. J Biosci Bioeng 98(5):317–330
Jin W, Brennan JD (2002) Properties and applications of proteins encapsulated within sol–gel derived materials. Anal Chim Acta 461(1):1–36
Labus K, Turek A, Liesiene J, Bryjak J (2011) Efficient Agaricus bisporus tyrosinase immobilization on cellulose-based carriers. Biochem Eng J 56(3):232–240
Lee N, Lee S-H, Baek K, Kim B-G (2015) Heterologous expression of tyrosinase (MelC2) from Streptomyces avermitilis MA4680 in E. coli and its application for ortho-hydroxylation of resveratrol to produce piceatannol. Appl Microbiol Biotechnol 99(19):7915–7924
Li X, Chen Z, Wu Y, Yan Y, Sun X, Yuan Q (2018) Establishing an artificial pathway for efficient biosynthesis of hydroxytyrosol. ACS Synth Biol 7(2):647–654
Li C, Jia P, Bai Y, Fan T-P, Zheng X, Cai Y (2019) Efficient synthesis of hydroxytyrosol from L-dopa using engineered Escherichia coli whole cells. J Agric Food Chem 67(24):6867–6873
Liebgott P-P, Labat M, Casalot L, Amouric A, Lorquin J (2007) Bioconversion of tyrosol into hydroxytyrosol and 3, 4-dihydroxyphenylacetic acid under hypersaline conditions by the new Halomonas sp. strain HTB24. FEMS Microbiol Lett 276(1):26–33
Liu D-M, Chen J, Shi Y-P (2018) Advances on methods and easy separated support materials for enzymes immobilization. TrAc Trend Anal Chem 102:332–342
Marín-Zamora ME, Rojas-Melgarejo F, García-Cánovas F, García-Ruiz PA (2009) Production of o-diphenols by immobilized mushroom tyrosinase. J Biotechnol 139(2):163–168
Min K, Park K, Park D-H, Yoo YJ (2015) Overview on the biotechnological production of L-DOPA. Appl Microbiol Biotechnol 99(2):575–584
Nolan LC, O’Connor KE (2007) Use of Pseudomonas mendocina, or recombinant Escherichia coli cells expressing toluene-4-monooxygenase, and a cell-free tyrosinase for the synthesis of 4-fluorocatechol from fluorobenzene. Biotechnol Lett 29(7):1045–1050
O’Connor K, Molloy S, Davis R, Shaw W (2017) A method for the enzymatic conversion of a phenol substrate into a corresponding catechol product. US 2017/0355969 A1
Orenes-Piñero E, García-Carmona F, Sánchez-Ferrer Á (2013) A new process for obtaining hydroxytyrosol using transformed Escherichia coli whole cells with phenol hydroxylase gene from Geobacillus thermoglucosidasius. Food Chem 139(1–4):377–383
Petrovičová T, Markošová K, Hegyi Z, Smonou I, Rosenberg M, Rebroš M (2018) Co-immobilization of ketoreductase and glucose dehydrogenase. Catalysts 8(4):168
Pierre A (2004) The sol-gel encapsulation of enzymes. Biocatal Biotransform 22(3):145–170
Reetz MT, Zonta A, Simpelkamp J (1996) Efficient immobilization of lipases by entrapment in hydrophobic sol-gel materials. Biotechnol Bioeng 49(5):527–534
Robles-Almazan M, Pulido-Moran M, Moreno-Fernandez J, Ramirez-Tortosa C, Rodriguez-Garcia C, Quiles JL, Ramirez-Tortosa M (2018) Hydroxytyrosol: bioavailability, toxicity, and clinical applications. Food Res Int 105:654–667
Rodríguez C, Lavandera I, Gotor V (2012) Recent advances in cofactor regeneration systems applied to biocatalyzed oxidative processes. Curr Org Chem 16(21):2525–2541
Santos M, Piccirillo C, Castro PM, Kalogerakis N, Pintado M (2012) Bioconversion of oleuropein to hydroxytyrosol by lactic acid bacteria. World J Microbiol Biotechnol 28(6):2435–2440
Shuster Ben-Yosef V, Fishman A (2009) Isolation, cloning and characterization of a tyrosinase with improved activity in organic solvents from Bacillus megaterium. J Mol Microbiol Biotechnol 17(4):188–200
Shuster Ben-Yosef V, Sendovski M, Fishman A (2010) Directed evolution of tyrosinase for enhanced monophenolase/diphenolase activity ratio. Enzyme Microb Technol 47(7):372–376
Tan D, Zhao J-P, Ran G-Q, Zhu X-L, Ding Y, Lu X-Y (2019) Highly efficient biocatalytic synthesis of L-DOPA using in situ immobilized Verrucomicrobiumspinosum tyrosinase on polyhydroxyalkanoate nano-granules. Appl Microbiol Biotechnol 103(14):5663–5678
Truppo M (2012) Cofactor recycling for enzyme catalyzed processes. In: Yamamoto H, Carreira EM (eds) Comprehensive chirality, vol 7. Elsevier, pp 46–70
Zdarta J, Meyer AS, Jesionowski T, Pinelo M (2018) Developments in support materials for immobilization of oxidoreductases: a comprehensive review. Adv Colloid Interf Sci 258:1–20
Zhang Z-L, Chen J, Xu Q, Rao C, Qiao C (2012) Efficient synthesis of hydroxytyrosol from 3, 4-dihydroxybenzaldehyde. Synth Commun 42(6):794–798
Zolghadri S, Bahrami A, Hassan Khan MT, Munoz-Munoz J, Garcia-Molina F, Garcia-Canovas F, Saboury AA (2019) A comprehensive review on tyrosinase inhibitors. J Enzyme Inhib Med Chem 34(1):279–309
Funding
This work was supported by the Israel Science Foundation founded by the Israel Academy of Sciences and Humanities, grant number 419/15. We also acknowledge the Russell-Berrie Nanotechnology Institute (RBNI) at the Technion for supporting this research. The authors are thankful for support from COST Action CM1303 (SysBiocat). MR is thankful to the Slovak Research and Development Agency contract no. APVV-16-0314 for their support.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
This article does not contain any studies with human participants or animals.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
ESM 1
(PDF 485 kb)
Rights and permissions
About this article
Cite this article
Deri-Zenaty, B., Bachar, S., Rebroš, M. et al. A coupled enzymatic reaction of tyrosinase and glucose dehydrogenase for the production of hydroxytyrosol. Appl Microbiol Biotechnol 104, 4945–4955 (2020). https://doi.org/10.1007/s00253-020-10594-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-020-10594-z