Isolation and Characterization of Thermophilic Cellulase-Producing Bacteria from Empty Fruit Bunches-Palm Oil Mill Effluent Compost

Abstract

Problems statement: Lack of information on locally isolated cellulase-producing bacterium in thermophilic compost using a mixture of Empty Fruit Bunch (EFB) and Palm Oil Mill Effluent (POME) as composting materials. Approach: The isolation of microbes from compost heap was conducted at day 7 of composting process where the mixture of composting materials consisted of 45.8% cellulose, 17.1% hemicellulose and 28.3% lignin content. The temperature, pH and moisture content of the composting pile at day 7 treatment were 58.3, 8.1 and 65.5°C, respectively. The morphological analysis of the isolated microbes was conducted using Scanning Electron Microscope (SEM) and Gram stain method. The congo red test was conducted in order to detect 1% CMC agar degradation activities. Total genomic DNAs were extracted from approximately 1.0 g of mixed compost and amplified by using PCR primers. The PCR product was sequent to identify the nearest relatives of 16S rRNA genes. The localization of bacteria chromosomes was determined by Fluorescence In Situ Hybridization (FISH) analysis. Results: Single isolated bacteria species was successfully isolated from Empty Fruit Bunch (EFB)-Palm Oil Mill Effluent (POME) compost at thermophilic stage. Restriction fragment length polymorphism profiles of the DNAs coding for the 16S rRNAs with the phylogenetic analysis showed that the isolated bacteria from EFB-POME thermophilic compost gave the highest homology (99%) with similarity to Geobacillus pallidus. The strain was spore forming bacteria and able to grow at 60°C with pH 7. Conclusion: Thermophilic bacteria strain, Geobacillus pallidus was successfully isolated from Empty Fruit Bunch (EFB) and Palm Oil Mil Effluent (POME) compost and characterized.

Microbial additives in the composting process

“…Therefore, it is possible that the inoculation of the compost with B. cereus increased the decomposition of that polymer. Some studies have reported the high cellulolytic potential of particular species of bacteria isolated from composting, such as those belonging to the genus Bacillus (Baharuddin;Razak;Hock, 2010;Lynd et al, 2012).…”

“…On the 23rd day (T10) the peaks associated with Treatments 3 and 4 were quite obvious in the region belonging to the inocula. The species of bacteria utilized as inocula are thermophiles, being plentiful in that phase of the process and together with fungi are responsible for cellulose degradation (Baharuddin;Razak;Hock, 2010;Lynd et al, 2012).Bacterias do genero Bacillus apresentam alta capacidade metabólica de degradar substâncias recalcitrantes como lignina e celulose, however. During the process, environmental conditions are mostly responsible for microbial fluctuations, and composting microbiota depends on its own competitive effectiveness and its survival capacity.…”

Characterization of thermostable cellulase produced by Bacillus strains isolated from solid waste of carrageenan

“…Bacteria which have high growth rate as compared to fungi have good potential to be used in cellulase production. For this application, the utilization of thermophilic cellulolytic microbes is more advantageous than the mesophilic one because the cellulase enzyme produced remains stable at high temperature and still has optimum activity [2]. Thermostable cellulase enzymes with high stability can be obtained by isolating from thermophilic bacteria.…”

Chitinolytic Bacteria Isolated from Chili Rhizosphere: Chitinase Characterization and Its Application as Biocontrol for Whitefly (<i>Bemisia tabaci</i> Genn.)

“…Amplification of the gene was carried out by PCR using Marchesi specific primer 63f (5'-CAG GCC TAA CAC ATG CAA GTC-3') and 1387r (5'-GGG CGG WGT GTA CAA GGC-3') (Wahyudi et al, 2010). The PCR condition was set up 30 cycles (Baharuddin et al, 2010) with pre-denaturing step at 94°C for 2 min, denaturing at 92°C for 30 sec, annealing at 55°C for 30 sec, polymerization at 75°C for 1 min and post PCR at 75°C for 5 min. The PCR products, approximately 1300 bp, were purified and sequenced using ABI 310 (Perkin Elmer, USA).…”