Chemical Papers

, Volume 73, Issue 4, pp 801–810 | Cite as

Preliminary assessment for the synthesis of lignin-type molecules using crude onion peroxidase

  • Laurentiu M. Palade
  • Constantin CroitoruEmail author
  • Anis Arnous
Original Paper


The aim of the study concerned the production of lignan-type molecules using inexpensive unpurified peroxidase from onion bulbs. The crude peroxidase (POD) extracted from onion bulbs is an inexpensive tool for the synthesis of fine chemicals as well as converting food industry waste materials into high added-value bio-products. The experiments were focusing on the ability of a crude onion POD preparation to act as a biocatalyst for the oxidative dimerization–cyclization of hydroxycinnamic acids and their methyl esters. The favorable conditions (pH and temperature) of the enzymatic reaction have been assayed using response surface methodology (RSM). Subsequently, a kinetics assay has been performed to distinguish the affinity of the enzyme towards the different substrates. The results indicated a strong substrate specificity toward methyl ferulate. Nevertheless, these findings on the ability of crude POD extract to generate dehydrodimers merit additional investigation and should be regarded as a preliminary assessment.


Food waste Onion peroxidase Hydroxycinnamates Optimization Response surface methodology 

List of symbols


Design of experiment


Substrate concentration at half the maximum reaction rate (µM)




Response surface methodology


Maximum rate of enzymatic reaction (µM min−1)



The present work is part of the Master Thesis of LMP elaborated at MAICh (CIHEAM). Special thanks to Anne Meyer from Denmark Technical University for the immense support. Heartfelt appreciation for Panagiotis Kefalas from MAICh (deceased). The work is dedicated to his memory.


  1. Arrieta-Baez D, Stark RE (2006) Modeling suberization with peroxidase-catalyzed polymerization of hydroxycinnamic acids: cross-coupling and dimerization reactions. Phytochemistry 67:743–753. CrossRefGoogle Scholar
  2. Arshad MS, Sohaib M, Nadeem M et al (2017) Status and trends of nutraceuticals from onion and onion by-products: a critical review. Cogent Food Agric 3:1–14. Google Scholar
  3. Battistuzzi G, Bellei M, Bortolotti CA, Sola M (2010) Redox properties of heme peroxidases. Arch Biochem Biophys 500:21–36. CrossRefGoogle Scholar
  4. Bezerra MA, Santelli RE, Oliveira EP et al (2008) Response surface methodology (RSM) as a tool for optimization in analytical chemistry. Talanta 76:965–977. CrossRefGoogle Scholar
  5. Burtis CA, Bostick WD, Overton JB, Mrochek JE (1981) Optimization of a kinetic method by response-surface methodology and centrifugal analysis and application to the enzymic measurement of ethanol. Anal Chem 53:1154–1159. CrossRefGoogle Scholar
  6. Cavazzuti M (2013) Design of experiments. In: Optimization methods: from theory to design scientific and technological aspects in mechanics. Springer, Berlin, pp 13–42.
  7. Chioccara F, Poli S, Rindone B et al (1993) Regio- and diastereo-selective synthesis of dimeric lignans using oxidative coupling. Acta Chem Scand 47:610–616CrossRefGoogle Scholar
  8. Cosio C, Dunand C (2009) Specific functions of individual class III peroxidase genes. J Exp Bot 60:391–408. CrossRefGoogle Scholar
  9. Daquino C, Rescifina A, Spatafora C, Tringali C (2009) Biomimetic synthesis of natural and “Unnatural” lignans by oxidative coupling of caffeic esters. Eur J Org Chem 2009:6289–6300. CrossRefGoogle Scholar
  10. de Montellano PRO (2010) In: Torres E, Ayala M (eds) Catalytic mechanisms of heme peroxidases BT—biocatalysis based on heme peroxidases: peroxidases as potential industrial biocatalysts. Springer, Berlin, Heidelberg, pp 79–107. CrossRefGoogle Scholar
  11. El Agha A, Makris DP, Kefalas P (2008a) Hydrocaffeic acid oxidation by a peroxidase homogenate from onion solid wastes. Eur Food Res Technol 227:1379–1386. CrossRefGoogle Scholar
  12. El Agha A, Makris DP, Kefalas P (2008b) Peroxidase-active cell free extract from onion solid wastes: biocatalytic properties and putative pathway of ferulic acid oxidation. J Biosci Bioeng 106:279–285. CrossRefGoogle Scholar
  13. El Agha A, Abbeddou S, Makris DP, Kefalas P (2009) Biocatalytic properties of a peroxidase-active cell-free extract from onion solid wastes: caffeic acid oxidation. Biodegradation 20:143–153. CrossRefGoogle Scholar
  14. El Ichi S, Miodek A, Sauriat-Dorizon H et al (2011) Characterization of structure and activity of garlic peroxidase (POX1B). J Biol Inorg Chem 16:157–172. CrossRefGoogle Scholar
  15. El-Seedi HR, El-Said AMA, Khalifa SAM et al (2012) Biosynthesis, natural sources, dietary intake, pharmacokinetic properties, and biological activities of hydroxycinnamic acids. J Agric Food Chem 60:10877–10895. CrossRefGoogle Scholar
  16. Esteves M, Siquet C, Gaspar A et al (2008) Antioxidant versus cytotoxic properties of hydroxycinnamic acid derivatives—a new paradigm in phenolic research. Arch Pharm (Weinheim) 341:164–173. CrossRefGoogle Scholar
  17. Fournier DA, Skaug HJ, Ancheta J et al (2012) AD model builder: using automatic differentiation for statistical inference of highly parameterized complex nonlinear models. Optim Methods Softw 27:233–249. CrossRefGoogle Scholar
  18. Frías I, Siverio JM, González C et al (1991) Purification of a new peroxidase catalysing the formation of lignan-type compounds. Biochem J 273:109–113CrossRefGoogle Scholar
  19. Garrido J, Gaspar A, Garrido EM et al (2012) Alkyl esters of hydroxycinnamic acids with improved antioxidant activity and lipophilicity protect PC12 cells against oxidative stress. Biochimie 94:961–967. CrossRefGoogle Scholar
  20. Girotto F, Alibardi L, Cossu R (2015) Food waste generation and industrial uses: a review. Waste Manag 45:32–41. CrossRefGoogle Scholar
  21. Hapiot P, Neudeck A, Pinson J et al (1996) Oxidation of caffeic acid and related hydroxycinnamic acids. J Electroanal Chem 405:169–176. CrossRefGoogle Scholar
  22. Hinkelmann K (2011) Design and analysis of experiments, special designs and applications. Wiley, HobokenGoogle Scholar
  23. Hynninen PH, Kaartinen V, Kolehmainen E (2010) Horseradish peroxidase-catalyzed oxidation of chlorophyll a with hydrogen peroxide Characterization of the products and mechanism of the reaction. Biochim Biophys Acta Bioenerg 1797:531–542. CrossRefGoogle Scholar
  24. Larsen E, Andreasen MF, Christensen LP (2001) Regioselective dimerization of ferulic acid in a micellar solution. J Agric Food Chem 49:3471–3475. CrossRefGoogle Scholar
  25. Liu H-L, Wan X, Huang X-F, Kong L-Y (2007) Biotransformation of sinapic acid catalyzed by Momordica charantia peroxidase. J Agric Food Chem 55:1003–1008. CrossRefGoogle Scholar
  26. Luis JC, Gonzalez FV, Perez RM et al (2005) Dimerization of ferulic and caffeic acids by purified peroxidase isolated from Bupleurum salicifolium callus culture. Prep Biochem Biotechnol 35:231–241. CrossRefGoogle Scholar
  27. Montgomery DC (1992) The use of statistical process control and design of experiments in product and process improvement. IIE Trans 24:4–17. CrossRefGoogle Scholar
  28. Motulsky H, Christopoulos A (2004) Fitting models to biological data using linear and nonlinear regression: a practical guide to curve fitting. Oxford University Press, OxfordGoogle Scholar
  29. Moural TW, Lewis KM, Barnaba C et al (2017) Characterization of class III peroxidases from switchgrass. Plant Physiol 173:417–433. 10.1104%2F pp. 16.01426CrossRefGoogle Scholar
  30. Moussouni S (2009) Crude peroxidase extract from onion : activity on O-diphenol & pentahydroxy chalcone oxidative cyclisation into aureusidin. MSc Thesis, Mediterranean Agronomic Institute of Chania (MAICh), Crete, GreeceGoogle Scholar
  31. Moussouni S, Detsi A, Majdalani M et al (2010) Crude peroxidase from onion solid waste as a tool for organic synthesis. Part I: cyclization of 2′,3,4,4′,6′-pentahydroxy-chalcone into aureusidin. Tetrahedron Lett 51:4076–4078. CrossRefGoogle Scholar
  32. Moussouni S, Saru M-L, Ioannou E et al (2011) Crude peroxidase from onion solid waste as a tool for organic synthesis. Part II: oxidative dimerization–cyclization of methyl p-coumarate, methyl caffeate and methyl ferulate. Tetrahedron Lett 52:1165–1168. CrossRefGoogle Scholar
  33. Myers RH, Montgomery DC, Anderson-Cook CM (2016) Response surface methodology: process and product optimization using designed experiments. Wiley, HobokenGoogle Scholar
  34. Orlandi M, Rindone B, Molteni G et al (2001) Asymmetric biomimetic oxidations of phenols: the mechanism of the diastereo- and enantioselective synthesis of dehydrodiconiferyl ferulate (DDF) and dehydrodiconiferyl alcohol (DDA). Tetrahedron 57:371–378. CrossRefGoogle Scholar
  35. Osman A, Makris DP, Kefalas P (2008) Investigation on biocatalytic properties of a peroxidase-active homogenate from onion solid wastes: an insight into quercetin oxidation mechanism. Process Biochem 43:861–867. CrossRefGoogle Scholar
  36. Osman A, El Agha A, Makris DP, Kefalas P (2012) Chlorogenic acid oxidation by a crude peroxidase preparation: biocatalytic characteristics and oxidation products. Food Bioprocess Technol 5:243–251. CrossRefGoogle Scholar
  37. Rai N, Yadav M, Yadav HS (2016) Enzymatic characterisation of lignin peroxidase from luffa aegyptiaca fruit juice. AM J Plant Sci 7:649–656CrossRefGoogle Scholar
  38. Ralph J, Garcia Conesa MT, Williamson G (1998) Simple preparation of 8 − 5-coupled diferulate. J Agric Food Chem 46:2531–2532. CrossRefGoogle Scholar
  39. Saliu F, Tolppa E-L, Zoia L, Orlandi M (2011) Horseradish peroxidase catalyzed oxidative cross-coupling reactions: the synthesis of ‘unnatural’ dihydrobenzofuran lignans. Tetrahedron Lett 52:3856–3860. CrossRefGoogle Scholar
  40. Setälä H, Pajunen A, Kilpeläinen I, Brunow G (1994) Horse radish peroxidase-catalysed oxidative coupling of methyl sinapate to give diastereoisomeric spiro dimers. J Chem Soc Perkin Trans 1:1163–1165. CrossRefGoogle Scholar
  41. Simpson D, Amos S (2017) Other Plant Metabolites. In: Delgoda R, Badal S (eds) Pharmacognosy. Fundamentals, applications and strategy. Academic Press, Boston, pp 267–280. Google Scholar
  42. Tafazoli S, O’Brien PJ (2005) Peroxidases: a role in the metabolism and side effects of drugs. Drug Discov Today 10:617–625. CrossRefGoogle Scholar
  43. Takahama U, Hirota S (2000) Deglucosidation of quercetin glucosides to the aglycone and formation of antifungal agents by peroxidase-dependent oxidation of quercetin on browning of onion scales. Plant Cell Physiol 41:1021–1029. CrossRefGoogle Scholar
  44. Villalobos DA, Buchanan ID (2002) Removal of aqueous phenol by Arthromyces ramosus peroxidase. J Environ Eng Sci 1:65–73. CrossRefGoogle Scholar
  45. Yu B-B, Han X-Z, Lou H-X (2007) Oligomers of resveratrol and ferulic acid prepared by peroxidase-catalyzed oxidation and their protective effects on cardiac injury. J Agric Food Chem 55:7753–7757. CrossRefGoogle Scholar
  46. Zámocký M, Hofbauer S, Schaffner I et al (2015) Independent evolution of four heme peroxidase superfamilies. Arch Biochem Biophys 574:108–119. CrossRefGoogle Scholar

Copyright information

© Institute of Chemistry, Slovak Academy of Sciences 2018

Authors and Affiliations

  1. 1.National Institute for Research and Development in Animal Biology and NutritionBalotestiRomania
  2. 2.University of Agronomic Science and Veterinary Medicine BucharestBucharestRomania
  3. 3.Bioterra UniversityBucharestRomania
  4. 4.Department of Food Sciences at The Academy of Agricultural and Forestry SciencesBucharestRomania
  5. 5.Center for BioProcess Engineering, Department of Chemical and Biochemical EngineeringTechnical University of DenmarkLyngbyDenmark
  6. 6.Glycom A/SHørsholmDenmark

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