Cloning of family 11 Xylanase, xynG1 from locally isolated aspergillus fumigatus RT-1 & its expression in escherichia coli

• Main Author: Md. Illias, Rosli
• Format: Conference or Workshop Item
• Published: 2009
• Subjects: QD Chemistry; QK Botany
• Online Access: http://eprints.utm.my/id/eprint/14979/; http://dms.library.utm.my:8080/vital/access/manager/Repository/vital:99635
• Institution: Universiti Teknologi Malaysia

Locally isolated fungi are screened for xylanase activity via Remazol Brilliant Blue-xylan agar plate for hydrolysis of 1,4-beta-D-xylosidic linkages and dinitrosalicylic acid (DNS) method for release of xylose. Two of the fungi which showed the highest xylanase activity is identified as Aspergillus fumigatus RT-1, with activity 8.1 U/ml, followed by Aspergillus fumigatus RT-2, with activity 7.8 U/mL. Family 11 xylanase consensus fragments are amplified from genomic DNA by PCR using consensus degenerate primers that exhibited the conserved motifs within fungal family 11 xylanase genes. Two full-length xylanase genes, named xlnA and xynG1 are obtained by PCR and RT-PCR using specific primers designed based on the sequences in the database. The full-length xlnA contained 739 bp, including one intron of 52 bp while the full-length xynG1 contained 712 bp, with one intron of 46 bp. The mature peptide of xynG1 is cloned into pET-21a and pET-22b and expressed in Escherichia coli BL21 (DE3).