A gene encoding a novel extremely thermostable 1,4-β-xylanase isolated directly from an environmental DNA sample

Anwar Sunna*, Peter L. Bergquist

*Corresponding author for this work

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Small-subunit (SSU) rRNA genes (rDNA) were amplified by PCR from a hot pool environmental DNA sample using Bacteria- or Archaea-specific rDNA primers. Unique rDNA types were identified by restriction fragment length polymorphism (RFLP) analysis and representative sequences were determined. Family 10 glycoside hydrolase consensus PCR primers were used to explore the occurrence and diversity of xylanase genes in the hot pool environmental DNA sample. Partial sequences for three different xylanases were obtained and genomic walking PCR (GWPCR), in combination with nested primer pairs, was used to obtained a unique 1,741-bp nucleotide sequence. Analysis of this sequence identified a putative XynA protein encoded by the xynA open reading frame. The single module novel xylanase shared sequence similarity to the family 10 glycoside hydrolases. The purified recombinant enzyme, XynA expressed in E. coli exhibited optimum activity at 100°C and pH 6.0, and was extremely thermostable at 90°C. The enzyme showed high specificity toward different xylans and xylooligosaccharides.

Original languageEnglish
Pages (from-to)63-70
Number of pages8
JournalExtremophiles
Volume7
Issue number1
DOIs
Publication statusPublished - 2003

    Fingerprint

Cite this