PERTANIKA JOURNAL OF SCIENCE AND TECHNOLOGY

The abundant availability of Leucaena leucocephala (petai belalang) in Malaysia was one of the contributing factors of this research. The phytochemicals obtained from this plant will contribute towards many fields mainly medicine and it becomes more favorable as compared to the modern medicines used these days. Diseases such as cancer, high blood pressure and diabetes are very crucial, and it occurs frequently among all ages of citizens in Malaysia. Therefore, phytochemicals in this plant can be a great substitute of modern medicines. However, lignin is known to interfere with the cell wall polysaccharides digestion, and hence considered as an anti-quality component in forages. To improve phytochemicals extraction, an effective degradation method of lignin is in demand. Thus, the goal of the present study was to investigate the feasibility of using enzymatic hydrolysis by laccase from Trametes versicolor (LTV) to breakdown lignin using mild reaction condition. To clarify the effect of enzymatic hydrolysis, a preliminary study on the percentage of lignin removal was conducted that taking the effect of LTV concentration, incubation temperature and time into consideration. The results were then used to analyze total flavonoid contents (TFC) of L.leucocephala seed after the extraction by using microwave-assisted hydrodistillation (MAHD) method. In view of this, extraction temperature and time were varied. TFC was estimated by UV-VIS spectrophotometer for quantification by using quercetin as standard. The result emerge from this study showed that the TFC was high in the extraction temperature of 60˚C in 12 min with an average of 1639 mgQE/g.d.w and the lowest TFC was recorded at 30˚C with 677 mgQE/g.d.w. Further analysis showed that enzymatic hydrolysis has assisted the extraction of flavonoids, thus, provided scientific method to extract flavonoid as an alternative therapy for various diseases.

• Azahar, N. F., Gani, S. S. A., & Mokhtar, N. F. M. (2017). Optimization of phenolics and flavonoids extraction conditions of Curcuma zedoaria leaves using response surface methodology. Chemistry Central Journal, 11(1), 1-10. https://doi.org/10.1186/s13065-017-0324-y

• Bustamante, J., van Stempvoort, S., García-Gallarreta, M., Houghton, J. A., Briers, H. K., Budarin, V. L., Matharu, A. S., & Clark, J. H. (2016). Microwave assisted hydro-distillation of essential oils from wet citrus peel waste. Journal of Cleaner Production, 137, 598-605. https://doi.org/10.1016/j.jclepro.2016.07.108

• de Carvalho M. E., Monteiro M. C., & Sant’ Anna, G. L. (1999). Laccase from Trametes versicolor: Stability at temperature and alkaline conditions and its effect on biobleaching of hardwood kraft pulp. Applied Biochemistry and Biotechnology, 77(79), 723-33. https://doi.org/10.1385/abab:79:1-3:723

• Dent, M., Dragović-Uzelac, V., Penić, M., Brñić, M., Bosiljkov, T., & Levaj, B. (2013). The effect of extraction solvents, temperature and time on the composition and mass fraction of polyphenols in dalmatian wild sage (Salvia officinalis L.) extracts. Food Technology and Biotechnology, 51(1), 84-91.

• Draganescu, D., Dodi, G., Stoica, I., & Popa, M. I. (2017). Hydrolysis studies of flaxseed extract by high performance liquid chromatography. Cellulose Chemistry and Technology, 51(9-10), 803-811.

• Farhat, A., Fabiano-Tixier, A. S., Maataoui, M. E., Maingonnat, J. F., Romdhane, M., & Chemat, F. (2011). Microwave steam diffusion for extraction of essential oil from orange peel: Kinetic data, extract’s global yield and mechanism. Food Chemistry, 125(1), 255-261. https://doi.org/10.1016/j.foodchem.2010.07.110

• Fukushima, Y., & Kirk, K. (1995). Laccase component of the Ceriporiopsis subvermispora lignin-degrading system. Applied and Environmental Microbiology, 61, 872-876.

• Gierer, J. (1985). Chemistry of delignification - Part 1: General concept and reactions during pulping. Wood Science and Technology, 19(4), 289-312.

• Hamidi, N. H. (2013) Enzymatic depolymerization of lignin by laccases (PhD thesis). University of Nottingham, UK.

• Hammed, A. M., Jaswir, I., Amid, A., Alam, Z., Asiyanbi-H, T. T., & Ramli, N. (2013). Enzymatic hydrolysis of plants and algae for extraction of bioactive compounds. Food Reviews International, 29(4), 352-370. https://doi.org/10.1080/87559129.2013.818012

• Hashmi, I., & Kim, J. G. (2003). Comparison of degradation studies in a biosimulator by continuous cultivation of Pseudomonas and indigenous microorganisms with varied dissolved oxygen concentrations. International Journal of Environment and Pollution, 19(2), 123-138. https://doi.org/10.1504/IJEP.2003.003744

• Ishmael, U. C., Rashid, S. S., & Ahmad, N. S. S. N. W. (2016, September 18). Optimization of laccase enzyme pretreatment process parameters of empty fruit bunches ( EFB ) using one-factor-at-a-time (OFAT). In Proceedings of the National Conference of Postgraduate Research 2016. Universiti Malaysia Pahang, Malaysia.

• Jeyaratnam, N., Nour, A. H., & Akindoyo, J. O. (2016). The potential of microwave assisted hydrodistillation in extraction of essential oil from Cinnamomum Cassia (cinnamon). ARPN Journal of Engineering and Applied Sciences, 11(4), 2179-2183.

• Kiranmai, M., Kumar, C. B. M., & Ibrahim, M. (2011). Comparison of total flavanoid content of Azadirachta indica root bark extracts prepared by different methods of extraction. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 2(3), 254-261.

• Kusuma, H. S., & Mahfud, M. (2017). Microwave-assisted hydrodistillation for extraction of essential oil from patchouli (Pogostemon cablin) leaves. Periodica Polytechnica Chemical Engineering, 61(2), 82-92. https://doi.org/10.3311/PPch.8676

• Markam, K. R. (1972). A novel flavone-polysaccharide compound from Monoclea forsteri. Phytochemistry, 11(6), 2047-2053. https://doi.org/10.1016/S0031-9422(00)90171-7.

• Moilanen, U., Kellock, M., Várnai, A., Andberg, M., & Viikari, L. (2014). Mechanisms of laccase-mediator treatments improving the enzymatic hydrolysis of pre-treated spruce. Biotechnology for Biofuels, 7(1), 1-13. https://doi.org/10.1186/s13068-014-0177-8

• Monteiro, M. C., & de Carvalho, M. E. (1998). Pulp bleaching using laccase from Trametes versicolor under high temperature and alkaline conditions. Applied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology, 983, 70-72. https://doi.org/10.1007/978-1-4612-1814-2_91

• Oyedemi, B. O., Oyedemi, S. O., Chibuzor, J. V, Ijeh, I. I., Coopoosamy, R. M., & Aiyegoro, A. O. (2018). Pharmacological evaluation of selected medicinal plants used in the management of oral and skin infections in Ebem-Ohafia District, Abia State, Nigeria. The Scientific World Journal, 2018, 1-16. https://doi.org/10.1155/2018/4757458

• Pérez, J., Muñoz-Dorado, J., De La Rubia, T., & Martínez, J. (2002). Biodegradation and biological treatments of cellulose, hemicellulose and lignin: An overview. International Microbiology, 5(2), 53-63. https://doi.org/10.1007/s10123-002-0062-3

• Pramod, S., Rao, K. S., & Sundberg, A. (2013). Structural, histochemical and chemical characterization of normal, tension and opposite wood of Subabul (Leucaena leucocephala (lam.) de wit.). Wood Science and Technology, 47(4), 777-796. https://doi.org/10.1007/s00226-013-0528-9

• Pytkowska, K., Bregisz, M., Majewski, S., Ratz-Łyko, A., & Arct, J. (2013). Effect of enzymatic hydrolysis on the antioxidant properties of alcoholic extracts of oilseed cakes. Food Technology and Biotechnology, 51(4), 539-546.

• Rachtanapun, P., Luangkamin, S., Tanprasert, K., & Suriyatem, R. (2012). Carboxymethyl cellulose film from durian rind. LWT – Food Science Technology 48(1), 52-58. https://doi.org/10.1016/j.lwt.2012.02.029

• Rezvanpanah, S., Rezaei, K., Razavi, S. H., & Moini, S. (2008). Use of microwave-assisted hydrodistillation to extract the essential oils from Satureja hortensis and Satureja montana. Food Science and Technology Research, 14(3), 311-314. https://doi.org/10.3136/fstr.14.311

• Slomczynski, D., Nakas, J. P., & Tanenbaum, S. W. (1995) Production and characterization of laccase from Botrytis cinerea. Applied and Environmental Microbiology, 6, 907-912.

• Tejado, A., Peña, C., Labidi, J., Echeverria, J. M., & Mondragon, I. (2007). Physico-chemical characterization of lignins from different sources for use in phenol-formaldehyde resin synthesis. Bioresource Technology, 98(8), 1655-1663. https://doi.org/10.1016/j.biortech.2006.05.042

• Valko, M., Leibfritz, D., Moncol, J., Cronin, M. T. D., Mazur, M., & Telser, J. (2007). Free radicals and antioxidants in normal physiological functions and human disease. International Journal of Biochemistry and Cell Biology, 39(1), 44-84. https://doi.org/10.1016/j.biocel.2006.07.001

• Yoswathana, N. (2013). Optimization of ScCO2 extraction of rambutan seed oil using response surface methodology. International Journal of Chemical Engineering and Applications, 4(4), 187-190. https://doi.org/10.7763/IJCEA.2013.V4.291