Malaysian Journal of Analytical Sciences Vol 24 No 4 (2020): 538 - 545

 

 

 

 

EVALUATION ON THE EFFICACY OF DIFFERENT DISRUPTION METHOD OF OIL PALM ROOTS AND Ganoderma boninense CELL WALL

 

(Penilaian ke atas Keberkesanan Kaedah Pemusnahan yang Berbeza pada Dinding Sel Akar Kelapa Sawit dan Ganoderma boninense)

 

Arnnyitte Alexander¹, Jedol Dayou², Coswald Stephen Sipaut³, Khim Phin-Chong¹*

 

¹Biotechnology Programme, Faculty of Science and Natural Resources

²Vibration and Sound Research Group (e-VIBS), Faculty of Science and Natural Resources

³Chemical Engineering Programme, Faculty of Engineering

Universiti Malaysia Sabah, 88400 Jalan UMS, Kota Kinabalu, Sabah, Malaysia

 

*Corresponding author:  chongkp@ums.edu.my

 

 

Received: 31 May 2020; Accepted: 7 July 2020; Published: 11 August 2020

 

 

Abstract

The cell wall is the outermost layer of cell encountered by pathogens, determining the infection fate. However, the chemical composition involves during host-pathogen interaction has not been fully elucidated. Prior to determining the composition in the cell walls, the walls must be isolated first from the intercellular component. Selected methods for oil palm roots and G. boninense cell disruption were evaluated for their suitability for cell wall isolation and presented in this paper. The effect of selected disruption methods on the content of crude proteins and DNA absorbance under 260 and 280 nm wavelengths in the produced cell wall materials was established on the basis of their solubilized materials. The investigated methods were; i) homogenization with sonication; ii) grinding with liquid nitrogen and; iii) lyophilization with homogenization. The cell wall materials with the highest degree of cytoplasmic component released were produced using lyophilization with homogenization. This was confirmed with the highest DNA absorbance at 260/280 nm of 2.25/1.95 for G. boninense and 2.47/2.35 for oil palm roots in their solubilized materials. The thus-produced preparation also released 4.5 and 10.1 μg/g of crude proteins for G. boninense and oil palm roots respectively. The degree of cell wall purification from intracellular components was validated using Fourier Transform Infrared Spectroscopy (FTIR). Isolated cell wall can be further utilized in cell wall composition analysis.

 

Keywords:  cell wall, disruption methods, lyophilization with homogenization

 

Abstrak

Dinding sel merupakan lapisan sel paling luar yang ditentang oleh patogen dan menentukan nasib jangkitan. Walau bagaimanapun, komposisi kimia yang terlibat semasa interaksi perumah-patogen masih lagi belum dijelaskan secara terperinci. Sebelum kajian ke atas komposisi dinding sel, dinding tersebut harus dipisahkan daripada komponen sel dalaman terlebih dahulu. Kesesuaian beberapa kaedah terpilih untuk memusnahkan sel akar kelapa sawit dan G. boninense telah dinilai untuk memisahkan dinding sel. Kesan kaedah pemusnahan sel terhadap kandungan protin dan penyerapan asid deoksiribonukleik di bawah panjang gelombang 260 dan 280 nm berdasarkan kepada bahan terlarut di dalam hasil bahan dinding sel telah dinilai. Kaedah pemusnahan yang dikaji adalah; i) homogenisasi dan sonikasi, ii) pengisaran dengan cecair nitrogen, iii) liofilisasi dan homogenisasi. Kaedah liofilisasi dan homogenisasi menghasilkan bahan dinding sel yang mempunyai komponen sitoplasma terbebas tertinggi. Keputusan ini disahkan dengan serapan asid deoksiribonukleik yang tinggi pada 260/280 nm di dalam bahan terlarut iaitu sebanyak 2.25/1.95 untuk G. boninense dan 2.47/2.35 untuk akar kelapa sawit. Selain itu, hasil dinding sel melalui keadah yang sama juga membebaskan 4.5 dan 10.1 μg/g protin masing-masing untuk G. boninense dan akar kelapa sawit. Tingkat kebersihan dinding sel daripada komponen dalaman telah disahkan dengan menggunakan spektrofotometer inframerah transformasi Fourier. Dinding sel yang terpisah seterusnya boleh digunakan di dalam analisa komposisi dinding sel.

 

Kata kunci:  dinding sel, kaedah gangguan, liofilisasi dan homogenisasi

 

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