Sains Malaysiana 50(12)(2021): 3547-3556

http://doi.org/10.17576/jsm-2021-5012-07

Pengoptimuman Campuran Enzim Selulase Rekombinan untuk Penguraian Tandan Kosong Kelapa Sawit

 (Optimization of Mixed Cellulase Enzymes for the Degradation of Oil Palm Empty Fruit Bunch)

SHAZILAH KAMARUDDIN*, FARAH DIBA ABU BAKAR & ABDUL MUNIR ABDUL MURAD

Department of Biological Sciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

Received: 28 September 2020/Accepted: 16 April 2021

ABSTRAK

Penggunaan enzim selulase untuk penguraian biojisim pertanian lignoselulosa telah lama dikaji dan pelbagai usaha telah dilakukan untuk meningkatkan kecekapan proses hidrolisis. Keberkesanan penguraian biojisim pertanian kepada gula ringkas memerlukan satu campuran enzim yang mengandungi pelbagai jenis aktiviti selulolitik. Dalam kajian ini, satu campuran multi-enzim rekombinan yang terdiri daripada tiga komponen asas selulase iaitu endoglukanase (EglB) dan β-glukosidase (BglA) daripada Aspergillus niger serta selobiohidrolase (CbhII) daripada Trichoderma virens telah dibentuk khusus untuk hidrolisis tandan kosong kelapa sawit (TKKS). Penghasilan enzim selulase rekombinan telah dilakukan menggunakan hos pengekspresan Pichia pastoris. Pengoptimuman nisbah enzim untuk tindak balas ditentukan menggunakan Kaedah Gerak Balas Permukaan (RSM). Hasil menunjukkan hidrolisis TKKS pada suhu 50 °C dan pH 5.0 menggunakan enzim pada nisbah 641.4 unit CMCase: 10.14 unit Avicelase: 93.8 unit β-glukosidase, menghasilkan gula terturun dan glukosa tertinggi, masing-masing sebanyak 63 mg dan 40 mg per gram substrat TKKS. Hasil hidrolisis TKKS oleh campuran multi enzim yang telah dibentuk dalam kajian ini menunjukkan ketiga-tiga gabungan enzim rekombinan ini berpotensi untuk digunakan bagi penguraian TKKS.

Kata kunci: Biojisim pertanian; hidrolisis enzim; kaedah gerak balas permukaan; koktel enzim

ABSTRACT

The use of cellulase enzymes in the degradation of lignocellulose agriculture biomass has long been studied and various efforts have been made to improve the efficiency of the hydrolysis process. The efficiency of enzymatic degradation of agricultural biomass to simple sugars requires a mixture of enzymes containing various types of cellulolytic activity. In this study, a recombinant multi-enzyme mixture consisting of three basic components of cellulase namely endoglucanase (EglB) and β-glucosidase (BglA) from Aspergillus niger as well as cellobiohydrolase (CbhII) of Trichoderma virens was created specifically for hydrolysis of oil palm empty fruit bunch (OPEFB). The production of recombinant cellulases has been performed using Pichia pastoris expression host. The enzyme ratio optimisation was determined using Response Surface Methodology (RSM). The results showed that the hydrolysis of OPEFB at 50 °C and pH 5.0 using enzymes at 641.4 units CMCase: 10.14 Avicelase units: 93.8 β-glucosidase units, produced the highest reducing sugar and glucose at 63 mg and 40 mg per gram of OPEFB substrate, respectively. The hydrolysis of OPEFB by a multi-enzyme mixture that has been formed in this study showed that these three combinations of recombinant enzymes have the potential to be used for the degradation of OPEFB.

Keywords: Agricultural biomass; enzymatic hydrolysis; enzyme cocktail; response surface methodology

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*Corresponding author; email: shazilah@ukm.edu.my