ISOLATION AND CHARACTERIZATION OF MANNANOLYTIC THERMOPHILIC BACTERIA FROM PALM OIL SHELL AND THEIR MANNANASE ENZYME PRODUCTION PROPERTIES
AbstractA mannanolytic thermophilic bacterium (L-07) was isolated from palm oil shell after 2 days ofenrichment in liquid medium supplemented with 1% palm kernel meal as mannan source. Sequenceanalysis of 16S-rRNA indicated that L-07 was similar (98%) toÂ Geobacillus stearothermophilus, aspecies of thermophilic aerobic bacteria. We found thatÂ G. stearothermophilus L-07 producedextracellular Î²-1,4-mannanases, but no Î²-manosidase and Î±-galactosidase activities. The growth of L-07reached its maximum (3.0 x 106Â cell/ml) at 12-20 hours, while the highestÂ Î²-mannanase activity (0.52U/ml) was observed in culture medium after 36 hours of cultivation at 60oC. The medium containinglocust bean gum was the best for producing extracellular Î²-1,4-mannanases compared with kolang kaling,konjak, and palm kernel meal. SDS-PAGE and zymogram analysis demonstrated that crude mannanasecomplex of L-07 from locust bean gum containing medium comprised three active bands with molecularweight of 85, 73 and 50 kDa.Â Â Keywords :Â Extracellular enzyme/mannanase/Geobacillus stearothermophilus
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