ISOLATION AND CHARACTERIZATION OF CELLULASE GENE Bacillus sp. RP1, ISOLATE FROM CIMANGGU HOTSPRING

• Main Author: Sardi, Arif
• Format: Theses
• Language: Indonesia
• Subjects: Ilmu hayati ; Biologi
• Online Access: https://digilib.itb.ac.id/gdl/view/36016
• Institution: Institut Teknologi Bandung

To move our economy onto a sustainable basis, it is essential that we find a replacement for fossil carbon as a source of liquid fuels and chemical industry feedstocks. Cellulose is the most abundant macromolecule on earth, has considerable be potential to contribute to addressing the issue. In addition to maintaining the continuity of energy, the use of this material will not cause competition with human food sources as occured when using starch for energy purpose. The efficient enzymatic hydrolysis of cellulose requires a coordinate and synergistic action of three groups of cellulases (endoglucanase, exoglucanase and ?-glucosidase). These enzymes are required to convert cellulose into glucose and then can be converted into ethanol as fuel substitute. Bacillus sp. RP1 is a thermophilic bacterium with huge potential for use in cellulose hydrolysis. This bacteria was isolated from hot springs Cimanggu allegedly can produce seven kinds of endoglucanases, six exoglucanases and four xylanases. This study aimed to isolate one of the cellulase gene from Bacillus sp. RP1. The research was begins by isolating exoglucanase gen from Bacillus sp. RP1, but was unable to obtain. It can happen due to several factors including lack of specific primers designed. There are several possible causes why the primers used could not recognize exoglucanase gene from Bacillus sp. RP1 i.e: (i) primer used is non specific to recognize exoglucanase gene, allegedly the area chosen as the primer design site is not a region (sequences) that truly conserved in the exoglucanase gene (there has been no report about motif of catalytic domain exoglucanase in bacteria/GH 48 specifically); (ii) sequences are used as reference do not represent exoglucanase genes from Bacillus sp. RP1 or Bacillus sp. RP1 has another type exoglucanase (instead of GH 48). In this research, has obtained a partial endoglucanase gene from Bacillus sp. Rp1 size of 876 bp. This gene has a very high homology (98.4%) with one of the genes endo-1,4-?-glucanase of B. subtilis subsp. subtilis str. 168. Although the gene obtained is a partial gene, expected that its activity will not be too much different from the intact endoglucanase gene. This is because the active sites of the protein is in the internal (midle) part of gene and still present in endoglucanase gene of Bacillus sp. Rp1 which have been isolated.