View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Universiti Teknologi Malaysia Institutional Repository HIGH CELL DENSITY CULTIVATION OF Hendersonia sp. FOR THE APPLICATION OF BIOLOGICAL CONTROL OF OIL PALM DISEASE MUHAMMAD DANIAL BIN AZMAN A dissertation submitted in partial fulfillment of the requirements for the award of the degree of Master of Engineering (Bioprocess) Faculty of Chemical Engineering Universiti Teknologi Malaysia JANUARY 2013 iii to my mom and dad for bringing me into this wonderful world iv ACKNOWLEDGEMENT Praise be upon Allah, the Almighty Who is Kind and Merciful. For it is by His grace and blessing that gave me the endurance and perseverance to complete this project, and ultimately, this dissertation. I would like to express my sincere gratitude to my supervisor, Prof. Dr. Hesham Ali El Enshasy, for giving me the opportunity to revive my long lost passion for research. There won’t be anything in the world that can compensate his endless support and advice. I also would like to acknowledge the help and support from everyone who was involved in this project, directly or indirectly, especially the staff members from the Insitute of Bioproduct Development (IBD). Once again, I wish to thank the research officers, Ms. Roslinda and Ms. Zalina, as well as Mr. Solleh and Mr. Yahya for sharing their knowledge and assistance. My deepest gratitude to all my fellow friends from the lab for their kindness and enormous help throughout the project. Working in the lab would never be fun and enjoyable without them around. Last but not least, I would like to thank my parents and family for their love and prayer. The success of this project would not be achieved if it were not for their continuous support. v ABSTRAK Malaysia merupakan salah satu pengeluar dan pengeksport produk kelapa sawit yang terbesar di dunia. Pada 2011, Malaysia telah mengeskport sebanyak 24.27 juta tan produk kelapa sawit yang membawa kepada nilai eksport sebanyak RM 80.4 billion. Walau bagaimanapun, pokok kelapa sawit mudah terdedah kepada penyakit, terutamanya Basal Stem Rot yang berpunca dari Ganoderma. Penyakit ini telah menyebabkan kerugian yang besar kepada ekonomi. Hendersonia sp. adalah strain kulat yang telah menunjukkan kesan efektif dalam menangani jangkitan Ganoderma. Kajian ini dijalankan bagi membina suatu platform pengkulturan Hendersonia yang komprehensif untuk pengeluaran skala industri. Bagi kajian screening medium agar, medium yang terbaik terdiri daripada (g/L): Ekstrak Malt, 20; Glukosa, 20; Pepton,1; Ekstrak Yis, 5; Serbuk Agar, 20. Bagi kajian screening shake flask pula, medium yang terdiri dari (g/L): Ekstrak Malta, 20; Glukosa, 20; Pepton,1; Ekstrak Yis, 5, memberi CDW yang tertinggi iaitu 7.45±0.6 g/L. Ini diikuti dengan medium yang terdiri dari of (g/L): Sukrosa, 40; KH2PO4, 1; MgSO4.7H2O, 1.0; KCl, 0.5; FeSO4, 0.01; Ekstrak Yis, 2, yang memberi CDW sebanyak 6.15±0.14 g/L. Optimasi medium telah dibuatkan dengan menggunakan Metodologi Response Surface. Formulasi medium yang telah dioptimasi terdiri dari (g/L): Sukrosa, 60; K2HPO4, 0.5; Ekstrak Yis, 3; MgSO4.7H2O, 1; KCl, 0.5; FeSO4, 0.01. Bagi pengkulturan bioreaktor 150-L, dengan menggunakan pH tanpa kawalan, CDW sebanyak 13.55 g/L telah diperolehi. Selain itu, kesan kelajuan agitasi kepada jisim sel telah dikaji di dalam bioreaktor 150-L. CDW yang tertinggi telah diperolehi dengan menggunakan kelajuan agitasi 250 rpm pada 70 jam. Kesimpulannya, kajian ini telah mencadangkan kaedah yang kos efektif dalam menghasilkan Hendersonia dalam skala industri. vi ABSTRACT Malaysia is one the world’s biggest producers and exporters of oil palm products. In 2011, Malaysia has exported 24.27 million tonnes of oil palm products, which accounted for RM 80.4 billion in total export revenue. However, the oil palm is susceptible to plant diseases, especially Basal Stem Rot caused by Ganoderma. This disease has caused tremendous losses to the economy. Hendersonia sp. is a novel fungus strain that has shown effective results in controlling Ganoderma infection. The aim of this study was to establish a comprehensive Hendersonia cultivation platform for industrial-scale production. In an agar medium screening study, the best medium was composed of (g/L): Malt Extract, 20; Glucose, 20; Peptone,1; Yeast Extract, 5; Agar powder, 20. Based from the shake flask media screening study, the medium composed of (g/L): Malt extract, 20; Glucose, 20; Peptone, 1; Yeast extract, 5, gave the highest CDW of 7.45±0.6 g/L. This is followed by the medium composed of (g/L): Sucrose, 40; KH2PO4, 1; MgSO4.7H2O, 1.0; KCl, 0.5; FeSO4, 0.01; Yeast Extract, 2, which resulted with CDW of 6.15±0.14 g/L. The optimization of the medium was applied by using Response Surface Methodology. The new optimized medium formulation was composed of (g/L): Sucrose, 60; K2HPO4, 0.5; Yeast Extract, 3; MgSO4.7H2O, 1; KCl, 0.5; FeSO4, 0.01. For the 150-L bioreactor cultivation, by utilizing un-controlled pH throughout the cultivation, 13.55 g/L of CDW was obtained. In addition, the effects of agitation speed on the cell mass during the 150-L bioreactor cultivation were studied. The highest CDW (15.6 g/L) was obtained with agitation speed of 250 rpm at 70 h. In conclusion, the present study has proposed a reliable and cost-effective approach in mass producing Hendersonia in industrial scale.