Malays. Appl. Biol. (2011) 40(1): 33-38
XYLANASE GENE FROM A LOCALLY ISOLATED BACTERIUM
HUSSAIN, M.H.*, CHONG, N.F.M., CHAN, C.S.W., SAFARINA, A. and HUSAINI, A.
Proteomics Laboratory, Department of Molecular Biology, Faculty of Resource Science and Technology, University Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia
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ABSTRACT
A xylanolytic bacterium was isolated from sago plantation humus. Results from the morphological observation, biochemical tests and 16s rRNA sequencing suggested the bacterium to be Klebsiella pneumoniae. Due to the xylanolytic activity of this bacterium, isolation and characterization of the xylanase gene were attempted. A distinct fragment of about 650 bp was successfully amplified using PCR and cloned into Escherichia coli XL-1 Blue. A BLAST search confirmed that the DNA sequence from the amplified fragment was endo-1,4-beta-xylanase gene from the family 11 glycoside hydrolase. It showed 98% homology with Bacillus subtilis xylanase gene. In silico characterisation showed an open reading frame encoding a 213 amino acid sequence with a molecular weight of 23.3 kDa and theoretical isolectric point (pI) at pH 9.42.
ABSTRAK
Bakteria pengurai xilan telah dipencilkan daripada sampel humus yang diambil di ladang sagu. Keputusan pemerhatian morfologi, ujian biokimia dan jujukan rRNA 16s mencadangkan bahawa bakteria ini ialah Klebsiella pneumoniae. Kajian juga dijalankan untuk memencil dan mencirikan gen xylanase daripada bakteria pengurai xilan ini. Produk bersaiz 650 pasangan bes telah berjaya diperolehi melalui proses PCR dan diklonkan ke dalam Escherichia coli XL-1 blue.. Analisis BLAST telah menunjukkan bahawa jujukan DNA produk tersebut merupakan gen endo-1,4-beta-xilanase daripada keluarga 11 glikosida hidrolase. Ia menunjukkan 98% kepadanan dengan gen xylanase Bacillus subtilis. Pencirian secara in silico menunjukkan ia mempuyai rangka bacaan terbuka (ORF) yang mengekodkan rantai peptida bersaiz 213 asid amino dengan berat molekul 23.3 kDa dan ramalan titik isoelektrik (pI) pada pH 9.42.
Key words: xylanase; sago; Klebsiella pneumoniae; humus.
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