Sequences analysis of gene encoding extracellular xylanase in Streptomyces costaricanus 45I-3

Abstract

Streptomyces costaricanus 45I-3 isolated from peat soil is a bacterial strain in the group of actinomycetes. This bacterium is known to produce extracellular xylanase. The objectives of this study were to analyze sequence of gene involved in the synthesis of extracellular xylanase. Complete gene encoding xylanase was isolated from the bacterial genome by Inverse Polymerase Chain Reaction (i-PCR). The resulted gene sequence in a total alignment of 1664 bp amplicons, which consisted of two open reading frame (ORF) with opposite direction. ORF1 consisted of 1029 bp and ORF2 (Partial sequence) consisted of 309 bp. BLASTX analysis showed ORF1 homology with xylanase of bacterium enrichment culture clone Xyl8B8 (GenBank accession No. AFH35005.1) that had 95% identity and 99% similarity. ORF2 homolog with glyoxalase bacterium enrichment culture clone Xyl8B8 (GenBank accession No. AFH35007.1) that had 95% identity and 98% similarity. Upstream of the ORF1 sequence was found Ribosome Binding Site (RBS) 7 bp from the start codon and putative promotor sequence at 100 bp (-35) and 77 bp (-10) from the start codon. Analysis of protein sequence deduced from ORF1 showed that there were 2 domains, i.e Glyco_hydrolase 11 (GH11) and Carbohydrate Binding Type 2 (CBM2). Active sites found on the 130 amino acid on GH11 domain. Visualization of 3 dimension structure showed that 1664 bp fragment had 19 area.