skip to main content

Enzymatic Phorbol Esters Degradation using the Germinated Jatropha Curcas Seed Lipase as Biocatalyst: Optimization Process Conditions by Response Surface Methodology

Graduate Program on Estate Crop Product Technology, Faculty of Agricultural Technology, Universitas Gadjah Mada, Jl. Flora, Bulaksumur, Yogyakarta 55281, Indonesia

Received: 29 Jun 2016; Published: 11 Oct 2016.
Open Access Copyright (c) 2016 by Authors, Published by BCREC Group under

Citation Format:
Cover Image

Utilization of Jatropha curcas seed cake is limited by the presence of phorbol esters (PE), which are the main toxic compound and heat stable. The objective of this research was to optimize the reaction conditions of the enzymatic PE degradation of the defatted Jatropha curcas seed cake (DJSC) using the acetone-dried lipase from the germinated Jatropha curcas seeds as a biocatalyst. Response Surface Methodology (RSM) using three-factors-three-levels Box-Behnken design was used to evaluate the effects of the reaction time, the ratio of buffer volume to DJSC, and the ratio of enzyme to DJSC on PE degradation. The results showed that the optimum conditions of PE degradation were 29.33 h, 51.11 : 6 (mL/g), and 30.10 : 5 (U/g cake) for the reaction time, the ratio of buffer volume to DJSC, and the ratio of enzyme to DJSC, respectively. The predicted degradation of PE was 98.96% and not significantly different with the validated data of PE degradation. PE content was 0.035 mg/g, in which it was lower than PE in non-toxic Jatropha seeds. The results indicated that enzymatic degradation of PE might be a promising method for degradation of PE. 

Fulltext View|Download
Keywords: jatropha seed cake; phorbol esters; enzymatic degradation; germinated jatropha lipase; response surface methodology

Article Metrics:

Article Info
Section: Original Research Articles
  1. Makkar, H.P.S., Becker, K. (2009). Jatropha curcas, a promising crop for the generation of biodiesel and value-added coproducts. European. Journal of Lipid Science and Technology, 1: 773-787
  2. Makkar, H.P.S., Aderibigbe, A.O., Becker, K. (1998). Comparative evaluation of nontoxic and toxic varieties of Jatropha curcas for chemical composition, digestibility, protein degradability and toxic factors. Food Chemistry, 62: 207-215
  3. Makkar, H.P.S., Becker, K., Sporer, F., Wink, M. (1997). Studies on nutritive potential and toxic constituents of different provenances of Jatropha curcas. Journal of Agricultural and Food Chemistry, 45: 3152-3157
  4. Aitken, A. (1986). The biochemical mechanism of action of phorbol esters. In: Evans F.J. (Ed.), Naturally Occurring Phorbol Esters. CRC Press, Boca Raton, pp. 271-288
  5. Devappa, R.K., Darukeshwara, J., Raj, K.R., Narasimhamurthy, K., Saibaba, P., Bhagya, S. (2008). Toxicity studies of detoxified Jatropha meal (Jatropha curcas) in rats. Food and Chemical Toxicology, 46: 3621-3625
  6. Martinez-Herrera, J., Siddhuraju, P., Francis, G., Davila-Ortiz, G., Becker, K. (2006). Chemical composition, toxic/antimetabolic constituents, and effects of different treatments on their levels, in four provenances of Jatropha curcas L. from Mexico. Food Chemistry, 96: 80-89
  7. Aregheore, E.M., Becker, K., Makkar, H.P.S. (2003). Detoxification of a toxic variety of Jatropha curcas using heat and chemical treatments, and preliminary nutritional evaluation with rats. South Pacific Journal of Natural Science, 21: 50-56
  8. Goel, G., Makkar, H.P.S., Francis, G., Becker, K. (2007). Phorbol esters: structure, biological activity, and toxicity in animals. International Journal of Toxicology, 26: 279-288
  9. Mentlein, R. (1986). The tumor promoter 12-O-tetradecanoyl phorbol 13-acetate and regulatory diacyl glycerols are substrates for the same carboxyl esterase. Journal of Biological Chemistry, 261: 7816-7818
  10. Cabot, M.C. (1985). Hydrolysis of novel diacylglycerol analogs and phorbol diesters by serum lipase. Biochemical and Biophysical Research Communications, 833(2): 330-335
  11. Kadner, S.S., Katz, J., Levitz, M., Finlay, T.H. (1985). The 65-kDa phorbol-diester hydrolase in mouse plasma is esterase 1 and is immunologically distinct from the 56-kDa phorbol-diester hydrolase in mouse liver. Journal of Biological Chemistry, 260: 15604-15609
  12. Fleming, N., Bilan, P.T., Sliwinski-Lis, E. (1986). Effects of a phorbol ester and diacylglycerols on secretion of mucin and arginine esterase by rat submandibular gland cells. Pflugers Arch, 406: 6-11
  13. Joshi, C., Mathur, P., Khare, S.K. (2011). Degradation of phorbol esters by Pseudomonas aeruginosa PseA during solid-state fermentation of deoiled jatropha curcas seed cake. Bioresource Technology, 102: 4815-4819
  14. de Barros, C.R.M., Ferreira, L.M.M., Nunes, F.M., Bezerra, R.M.F., Dias, A.A., Guedes, C.V., Cone, J.W., Marques, G.S.M., Rodrigues, M.A.M. (2011). The potential of white-rot fungi to degrade phorbol esters of Jatropha curcas L. seed cake. Engineering in Life Sciences, 11(1): 107-110
  15. Belewu, M.A., Belewu, K.Y., Ogunsola, F.O. (2010). Nutritive value of dietary fungi treated jatropha curcas kernel cake: Voluntary intake, growth and digestibility coefficient of goat. Agriculture and Biology Journal of North America, 1(2): 135-138
  16. Hidayat, C., Hastuti, P., Wardhani, A.K., Nadia, L.S. (2014). Method of phorbol ester degradation in Jatropha curcas L. seed cake using rice bran lipase. Journal of Bioscience and Bioengineering, 177(3): 372-374
  17. Hidayat C., Hastuti P., Utazmi S., Wardhani A.K., Pradipta D.S. (2014). Enhancing indigenous lipase activity of germinated Jatropha curcas L. seeds for the enzymatic degradation of phorbol ester. Biocatalysis and Agricultural Biotechnology, 3(3): 71-76
  18. Adebowale, Y.A., Adebowale, K.O. (2008). Emulsifying property of mucuna flour and protein isolates. Journal of Food Technology, 6(2): 66-79
  19. Saetae, D., Suntornsuk, W. (2010). Antifungal activities of ethanolic extract from jatropha curcas seed cake. Journal of Microbiology and Biotechnology, 20: 319-324
  20. Demissie, A.G., Lele, S.S. (2010). Bioassay-assisted identification of phorbol ester from Jatropha curcas (Linn.) tissue culture. International Journal of Pharma and Bio Sciences, 1: 1-7
  21. Zulkali, M.M.D., Ahmad, A.L., Norulakmal, N.H. (2006). Oryza sativa L. husk as heavy metal adsorbent: Optimization with lead as model solution. Bioresource Technology, 97: 21-25

Last update: 2021-10-14 21:43:10

No citation recorded.

Last update: 2021-10-14 21:43:10

  1. Comparative study between chemical, physical and enzymatic methods for Jatropha curcas kernel meal phorbol ester detoxification

    Abou-Arab A.. Heliyon, 5 (5), 2019. doi: 10.1016/j.heliyon.2019.e01689
  2. Modelling and analysis of critical factors for phorbol ester degradation in Jatropha seed cake using ISM and MICMAC

    Ahluwalia S.. International Journal of Environment and Waste Management, 21 (4), 2018. doi: 10.1504/IJEWM.2018.093437
  3. Lipases: From Production to Applications

    Melani N.B.. Separation and Purification Reviews, 49 (2), 2020. doi: 10.1080/15422119.2018.1564328