Open AccessDirect Cloning of a Xylanase Gene from Pawan-Riau Hot Spring
Author(s) -
Is Helianti
Publication year - 2007
Publication title -
hayati journal of biosciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.305
H-Index - 17
eISSN - 2086-4094
pISSN - 1978-3019
DOI - 10.4308/hjb.14.2.54
Subject(s) - open reading frame , gene , cloning (programming) , recombinant dna , xylanase , bacillus subtilis , biology , homology (biology) , expression vector , nucleic acid sequence , microbiology and biotechnology , molecular cloning , amino acid , peptide sequence , biochemistry , genetics , enzyme , bacteria , computer science , programming language
A functional gene containing an Open Reading Frame (ORF) encoding a β-1, 4-endoxylanase glycosyl hydrolase family 11 was cloned directly using metagenomic PCR-cloning method from Pawan Hot Spring sample in Riau. The gene consisted of 642 nucleotides, encoded for 213 amino acids. The amino acid sequence analysis using BLAST showed that the gene has high homology (93%) with xylanase gene from Bacillus subtilis. The gene showed its function when it was subcloned into an expression vector and overexpressed in E. coli. The crude extract of the recombinant enzyme had activity for 170 U/ml at 50 °C. The result of this work showed that metagenomic approach was a powerful short cut method to obtain recombinant biocatalyst that was useful for industrial application