2264 11 archive 2 disk0/00/00/22/64 2008-07-03 06:42:41 2008-07-03 06:42:41 2008-07-03 06:42:41 article show Nik Marzuki Sidik nms@pkrisc.cc.ukm.my Shaiful Adzni Sharifuddin Sahidan Senafi Universiti Kebangsaan Malaysia. Faculty of Science and Technology. School of Bioscience and Biotechnology Malaysian Argricultural Research and Development Institute. Biotechnology Center, Universiti Kebangsaan Malaysia. Faculty of Science and Technology. School of Bioscience and Biotechnology pub Q R none Aspergillus terreus SUK-1, cellulase, β-glucosidase, PCR This study was funded by an IRPA grant 09-02-02-0108-EA260, Ministry of Science, Technology and Innovation, Malaysia. The breakdown of cellulose to glucose is catalysed by a complex cellulase enzyme which is comprised of three classes of enzymes; endoglucanase, exoglucanase and β-glucosidase. Many studies have been done on cellulase and several genes encoding for β-glucosidase have been reported. Here we describe the cloning and sequence analysis of partial putative β-glucosidase gene from Aspergillus terreus SUK-1. Its potential to produce cellulase enzyme has been studied and reported. A pair of specific primer was designed from the conserved regions of several fungal sequences taken out from the GenBank. PCR amplification has successfully amplified a DNA fragment of approximately 233 bp and this fragment was cloned (designated as SA3). The sequence analysis with BLAST showed high similarity towards β-glucosidase gene and protein. 80% and 79% homology were observed between SA3 nucleotide sequence and β-glucosidase gene of A. niger and both A. kawachii and A. aculeatus. Whereas, amino acid sequence of SA3 showed 83% homology towards β-glucosidase of A. niger, A. kawachii and A. aculeatus. 2005 published Malaysian Journal of Biochemistry and Molecular Biology 12 1 Malaysian Society for Biochemistry and Molecular Biology 61-63 TRUE ISSN 1511-2616 http://ejum.fsktm.um.edu.my/ArticleInformation.aspx?ArticleID=614 1. Barnett C, Berka R, Fowler T. Cloning and amplification of the gene encoding an extracellular β-glucosidase from Trichoderma reesei: evidence for improved rates of saccharification of cellulosic substrates. Bio/Technol 1991; 9: 562-567. 2. Silva J, Saloheimo M, Montanez C,Knowles JKC, Teeri TT. Molecular cloning of cellulolytic enzyme genes from References Cellulomonas flavigena in E. coli. J Biotechnol 1988; 8:249-256. 3. Nakamura K, Misawa N, Kitomura K. Cellulase genes of Cellulomonas uda CB4. I. Cloning and expression of β-glucosidase genes in E. coli. J Biotechnol 1986; 3: 239-246. 4. Yablonsky MD, Bartley T, Elliston KD, Kahrs, SK, Shalita, ZP, Eveleigh, DE. 1988. Characterization and cloning of cellulase complex of Microbispora bispora. In: Aubert, J.-P., Beguin, P. and Millet, J. (Eds.), Biochemistry and Genetics of Cellulosic Degradation. FEMS Symphosium No. 43, Academic Pres, London, 1988; pp 249-266. 5. Millet J, Pétrè D, Béguin P, Raynaud O, and Aubert JP. Cloning of ten distinct DNA fragments of Clostridium thermocellum coding for cellulases. FEMS Microbiol Let 1985; 29: 145-149. 6. Kawaguchi T, Enoki T., Tsurumaki S, Sumitami J, Ueda M, Ooi T, Arai M. Cloning and sequencing of the cDNA encoding β-glucosidase 1 from Aspergillus aculeatus. Gene 1996; 173: 287-288. 7. Omar O, Kader J, Al-Arikah Raja-E Ali. Studies on extracellular cellulases during the growth of Aspergillus SUK-1. Sains Malaysiana 1984; 13: 341-354. 8. Sambrook J, Fritsch E, Maniatis T. Molecular cloning: In a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, New York, 1989.