An enhanced fat extraction from Pycnanthus angolensis (African nutmeg) seeds using cellulase from Aspergillus niger strain BC23

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Marius Obiwanne Chiwetalu
Emmanuel Chekwube Ossai
Osmond Chukwuma Enechi
Christian Nnabuchi Ugwu
Vincent E.O. Ozogwu
Charles Odilichukwu R. Okpala
Obioma Uzoma Njoku


acid value, Aspergillus niger, cellulase, peroxide value, Pycnanthus angolensis, saponification


A promising extraction method for obtaining fat from Pycnanthus angolensis seeds through Aspergillus niger strain BC23 enzyme pretreatment and particle size reduction is presented in this work. The employed A. niger enzyme was produced from corn bagasse waste material, which served as the only source of carbon. P. angolensis seeds were dried, pulverized and subsequently sieved into two different particle sizes of 1.0 mm and 1.4 mm. This was followed by enzyme pretreatment prior to fat extraction. Results demonstrated that while the peak enzyme activity (121.036 U/mL) occurred on day 4 of fermentation, the maximum cellulase activity could be achieved at 60% saturation of ammonium sulfate purification. Compared with the results of water-treated and untreated seed samples, the fat yield, free fatty acid and iodine values of the enzyme-treated seed samples were considerably higher (P < 0.05). As peroxide value was less in the enzyme-assisted extracted fat, which pointed to a good-quality oil, the decreased refractive index suggested peak unsaturation. Despite considerable differences (P < 0.05) in refractive indices, fat’s specific gravity matched (P > 0.05) when enzyme-treated, water-treated and untreated samples were compared. On the whole, size reduction of P. angolensis seeds pretreated with A. niger cellulase enzyme to improve both fat quality and yield implied it as a useful inventory for confectionary as well as cosmetic industries.

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Abubakar F.A. and Oloyede O.B., 2013. Production and activity of cellulase from Aspergillus niger using rice bran and orange peel as substrates. International Journal of Scientific Research and Management 1(5): 285–291.

Aliyah, A., Alamsyah, G., Ramadhani, R. and Hermansyah, H., 2017. Production of α-amylase and β-glucosidase from Aspergillus niger by solid state fermentation method on biomass waste substrates from rice husk, bagasse and corn cob. Energy Procedia 136, 418–423. 10.1016/j.egypro.2017.10.269

Ashori, A., Nourbakhsh, A. and Tabrizi, A.K., 2014. Thermoplastic hybrid composites using bagasse, corn stalk and E-glass fibers: fabrication and characterization. Polymer-Plastics Technology and Engineering, 53(1), 1–8. 10.1080/03602559.2013.832854

Association of Official Analytical Chemists (AOAC). 2005. Official methods of analysis, 18th edition. AOAC, MD, USA.

Atoyebi, O.D., Osueke, C.O., Badiru, S., Gana, A.J., Ikpotokin, I., Modupe, A.E. and Tegene, G.A., 2019. Evaluation of particle board from sugarcane bagasse and corn cob. International Journal of Mechanical Engineering and Technology (IJMET) 10(1), 1193–1200.

Bayisa Y.M. and Bullo T.A., 2021. Optimization and characterization of oil extracted from Crotonmacrostachyus seed for antimicrobial activity using experimental analysis of variance. Heliyon 7: e08095. 10.1016/j.heliyon.2021.e08095

Borawski A.B., Borawski P. and Dunn J.W., 2016. Factors of the development of oilseed rape production in Poland on the background of the world. Journal of Advanced Agricultural Technologies 3(3): 175–179. 10.18178/joaat.3.3.175–179

Bradner J.R., Gillings M. and Nevalainen K.M.H., 1999. Qualitative assessment of hydrolytic activities in Antarctic micro fungi grown at different temperatures on solid media. World Journal Microbiology and Biotechnology 15: 131–132. 10.1023/A:1008855406319

Cecilia J.A., Plata D.B., Saboya R.M.A., Luna F.M.T., Cavalcante C.L., Rodriguez-Castellon E., 2020. An overview of the biolubri-cant production process: Challenges and future perspectives. Processes 8: 257. 10.3390/pr8030257

Costa P.C., Reis E.E, Carvalho L.S., Fontana R.C., Schneider W.D.H. and Camassola M., 2021. Making the process of enzyme production in solid-state cultivation cleaner and more sustainable—reuse of raw materials and a syringe-type bioreactor enter in the scene. ACS Sustainable Chemistry & Engineering 42, 14134–14142. 10.1021/acssuschemeng.1c04257

Dash D.R., Pathak S.S. and Pradhan R.C., 2021. Extraction of oil from Terminalia chebulakernel by using ultrasound technology: influence of process parameters on extraction kinetics. Industrial Crops & Products 171: 113893. 10.1016/j.indcrop.2021.113893

Díaz-Suárez P., Rosales-Quintero A., Fernandez-Lafuente R., Pola-Sanchez E., Hernandez-Cruz M.C., Ovando-Chcon S.L., Rodrigues R.C. and Tacias-Pascacio V.G., 2021. Aqueous enzymatic extraction of Ricinuscommunis seeds oil using Viscozyme L. Industrial Crops and Products 170: 113811. 10.1016/j.indcrop.2021.113811

Didia B., Zakpaa H.D., Mills-Robertson F.C. and Abdul-Mumeen I, 2018. Qualitative analysis of enzyme-assisted traditionally extracted shea butter from different pretreated shea kernel substrates. Journal of Cereals and Oilseeds 9(2): 10–19. 10.5897/JCO2017.0177

Elagbar Z.A., Naik R.R., Shakya A.K. and Bardaweel S.K., 2016. Fatty acids analysis, antioxidant and biological activity of fixed oil of Annonamuricata L. seeds. Journal of Chemistry. 2016: ID 6948098. 10.1155/2016/6948098

Gilna VV. and Khaleel K.M., 2011. Cellulase enzyme activity of Aspergillus fumigatesfrom mangrove soil on lignocellulosic substrate. Recent Research in Science and Technology 3(1): 132–134.

Gregg, D.J. and Saddler, J.N., 1996. Factors affecting cellulose hydrolysis and the potential of enzyme recycle to enhance the efficiency of an integrated wood to ethanol process. Biotechnology and Bioengineering 51(4), 375–383.

Gustafson K., Wu Q.L., Asante-Dartey J. and Simon J.E., 2013. Pycnanthus angolensis: bioactive compounds and medicinal applications. In: Juliani H.R., Simon J.E. and Ho C.T. (eds.). African natural plant products, vol II: Discoveries and challenges in chemistry, health and nutrition. ACS Sumposium Series, Ashington DC, pp. 63–78. 10.1021/bk-2013-1127.ch005

He Y., Zhang B., Guo S., Guo Z., Chen B. and Wang M., 2020. Sustainable biodiesel production from the green microalgae Nannochloropsis: novel integrated processes from cultivation to enzyme-assisted extraction and ethanolysis of lipids. Energy Conversion and Management 209: 112618. 10.1016/j.enconman.2020.112618

Ibeto C.N., Okoye C.O.B. and Ofoefule A.U., 2012. Comparative study of the physicochemical characterization of some oils as potential feedstock for biodiesel production. ISRN Renewable Energy 2012: ID 621518. 10.5402/2012/621518

Ikhuoria E.U. and Maliki M., 2007. Characterization of avocado pear (Persea americana) and African pear (Dacryodes edulis) extract. African Journal of Biotechnology 7: 950–952.

Ishak I., Hussain N., Coorey R. and Ghani M.A., 2021. Optimization and characterization of chia seed (Salvia hispanica L.) oil extraction using supercritical carbon dioxide. Journal of CO2 Utilization 45: 101430. 10.1016/j.jcou.2020.101430

Kalia V.C., Rashmi L.S., and Gupta M.N., 2001. Using enzymes for oil recovery from edible seeds. Journal of Scientific & Industrial Research 60: 298–310.

Kumar R.R., Bhargava D.V., Pandit K., Goswami S., Shankar S.M., Singh S.P., Rai G.K., Satyavathi C.T. and Praveen S., 2021. Lipase—the fascinating dynamics of enzyme in seed storage and germination—a real challenge to pearl millet. Food Chemistry 361: 130031. 10.1016/j.foodchem.2021.130031

Li F., Yang L., Zhao T., Zhao J., Zou Y. and Wu X., 2012. Optimization of the enzymatic pretreatment for n-hexane extraction of oil from Silybum marianum seed using response surface methodology. Food and Bioproduct Processing 90(2): 87–94. 10.1016/j.fbp.2011.02.010

Liu J., Gasmalla M.A.A., Li P. and Yang R., 2016. Enzyme-assisted extraction processing from oilseeds: principle, processing and application. Innovative Food Science and Emerging Technologies 35: 184–193. 10.1016/j.ifset.2016.05.002

Liu Z., Gui M., Xu T., Zhang L., Kong L., Qin L. and Zou Z., 2019. Efficient aqueous enzymatic-ultrasonication extraction of oil from Sapindusmukorossi seed kernels. Industrial Crops & Products 134: 124–133. 10.1016/j.indcrop.2019.03.065

Lourith N., Kanlayavattanakul M., Mongkonpaibool K., Butsaratrakool T. and Chinmuang T., 2016. Rambutan seed as a new promising unconventional source of specialty fat for cosmetics. Industrial Crops and Products 83: 149–154. 10.1016/j.indcrop.2015.12.045

Lowry O.H., Rosebrough N.J., Farr A.L. and Randall R.J., 1951. Protein measurement with the folin phenol reagent. Journal of Biology and Chemistry 193: 265–275.

Maftukhah S. and Abdullah A., 2018. Cellulase enzyme production from rice straw using solid state fermentation and fungi Aspergillus niger ITBCC L74. MATEC Web of Conferences 156, 01010. 10.1051/matecconf/201815601010

Mapongmetsem P.M., 2007. Pycnanthus angolensis (Welw.) Warb. In: Vossen H.A.M. and Mkamilo G.S. (editors). Plant resources of tropical Africa, 14: Vegetable oils. Backhuys, Wageningen, the Netherlands, pp. 137–141.

Mohammed N.K., Samir Z.T., Jassim M.A. and Saeed S.K., 2021. Effect of different extraction methods on physicochemical properties, antioxidant activity, of virgin coconut oil. Materials Today: Proceedings 42: 2000–2005. 10.1016/j.matpr.2020.12.248

Mu J., Wu G., Chen Z., Brennan C.S., Tran K., Dilrukshi H.N.N., Shi C., Zhen H. and Hui X., 2021. Identification of the fatty acids profiles in supercritical CO2 fluid and Soxhlet extraction of Samara oil from different cultivars of Elaeagnus mollis Diels seeds. Journal of Food Composition and Analysis 101: 103982. 10.1016/j.jfca.2021.103982

Nagre R.D., Oduro I. and Ellis W.O., 2011. Comparative physicochemical evaluation of kombo kernel fat produced by three different processes. African Journal of Food Science and Technology 2(4): 83–91.

Nandi S. and Bhattacharyya R., 2015. A parametric study for the enzymatic extraction of rice bran oil and its optimization. International Journal on Recent and Innovation Trends in Computing and Communication 3 (2): 21–25.

Nhi T.T.Y. and Tuan D.Q., 2016. Enzyme-assisted extraction of gac oil (Momordica cochinchinensis Spreng) from dried aril. Journal of Food and Nutrition Sciences 4(1): 1–6. 10.11648/j.jfns.20160401.11

Okoye I.G., Ezugwu A.L., Udenwobele D.I., Eze S.O.O., Anyawu C.U. and Chilaka F.C., 2013. Production and partial characterization of cellulases from Apergillus fumigates using two distinct parts of corn cob as carbon sources. Nigerian Journal of Biotechechnology 26: 50–59.

Otu S.A., Dzogbefia V.P., Kpikpi E.N. and Essuman E.K., 2015. Comparative effect of crude and commercial enzyme in shea fat extraction. IOSR Journal of Biotechnology and Biochemistry 1(3): 18–27.

Patil S.R. and Dayanand A., 2006. Production of pectinase from deseeded sunflower head by Aspergillus niger in submerged and solid-state conditions. Bioresources Technology 97: 2054–2058. 10.1016/j.biortech.2005.09.015

Pitt J.I. and Hocking A.D., 2009. Aspergillus and related teleo-morphs. In: Fungi and food spoilage, Pitt J.I. and Hocking A.D., Editors, Springer, Boston, MA, pp. 275–337. 10.1007/978-0-387-92207-2_8

Pointing S.P., 1999. Qualitative methods for the determination of lignocellulolytic enzyme production by tropical fungi. Fungal Diversity 2: 17–33.

Qiu C., He Y., Huang Z., Li S., Huang J., Wang M. and Chen B., 2019. Lipid extraction from wet nannochloropsis biomass via enzyme-assisted three phase partitioning. Bioresource Technology 284: 381–390. 10.1016/j.biortech.2019.03.148

Salina H.F., Shima A.R.R., Masniza M. and Faeizah H.N., 2013. Enzyme-assisted aqueous extraction and phenolic antioxidants of onions oil. International Journal of Science, Environment and Technology 2(5): 949–955.

Shivasharanappa K., Hanchinalmath J.V., Sundeep Y.S., Borah D. and Talluri V.P. 2014. Optimization and production of alkaline proteases from agro byproducts using a novel trichoderma viridiae strain VPG 12, isolated from agro soil. International Letters of Natural Sciences 14: 77–84. 10.18052/

Sirisompong W., Jirapakkul W. and Klinkesorn U., 2011. Response surface optimization and characteristics of rambutan (Nepheliumlappaceum L.) kernel fat by hexane extraction. LWT–Food Science and Technology 44: 1946–1951. 10.1016/j.lwt.2011.04.011

Sofidiya M.O. and Awolesi A.O., 2015. Antinociceptive and antiulcer activities of Pycnanthus angolensis. Revista Brasileira de Farmacognosia 25: 252–257. 10.1016/j.bjp.2015.05.004

Srivastava N., Singh R., Mohammad A., Pal D.B., Syed A., Elgorban A.M., Mishra P.K., Yoon T., Srivastava M. and Gupta V.K., 2022. Graphene oxide mediated enhanced cellulase production using pomegranate waste following co-cultured condition with improved pH and thermal stability. Fuel 312: 122807. 10.1016/j.fuel.2021.122807

Srivastava N., Srivastava, M., Manikanta A., Singh P., Ramteke P.W., Mishra P.K. and Malhotra B.D., 2017. Production and optimization of physicochemical parameters of cellulase using untreated orange waste by newly isolated Emericella variecolor NS3. Applied Biochemistry and Biotechnology 183(2): 601–612. 10.1007/s12010-017-2561-x

Talkit K.M., Mahajan D.T. and Masand V.H., 2012. Study on physicochemical properties of vegetable oils and their blends use as possible ecological lubricant. Journal of Chemical and Pharmaceutical Research 4(12): 5139–5144.

Tano-Deborah K. and Ohta Y., 1994. Enzyme-assisted aqueous extraction of fat from kernels of the shea tree, Butyrospermum parkii. Journal of the American Oil Chemists’ Society 71: 979–983. 10.1007/BF02542265

Ullah J., Hamayoun M., Ahmad T., Ayub M. and Zafarullah M., 2003. Effect of light, natural and synthetic antioxidants on stability of edible oil and fats. Asian Journal of Plant Sciences 2: 1192–1194. 10.3923/ajps.2003.1192.1194

Zhang Y., Kong X., Wang Z., Sun Y., Zhu S., Li L. and Lv P., 2018. Optimization of enzymatic hydrolysis for effective lipid extraction from microalgae Scenedesmus sp. Renewable Energy 125: 1049–1057. 10.1016/j.renene.2018.01.078

Zhang S.B., Lu Q.Y., Yang H., Li Y. and Wang S., 2011. Aqueous enzymatic extraction of oil and protein hydrolysates from roasted peanut seeds. Journal of American Oil Chemists’ Society 88(5): 727–732. 10.1007/s11746-010-1711-x

Zhang C., Zhao J., Famous E., Pan S., Peng X. and Tian J., 2021. Antioxidant, hepatoprotective and antifungal activities of black pepper (Piper nigrum L.) essential oil. Food Chemistry 346: 128845. 10.1016/j.foodchem.2020.128845