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  • Title: Overexpression and single-step purification of a thermostable xylanase from Bacillus stearothermophilus T-6.
    Author: Lapidot A, Mechaly A, Shoham Y.
    Journal: J Biotechnol; 1996 Nov 15; 51(3):259-64. PubMed ID: 8988650.
    Abstract:
    Xylanase T-6 is a thermostable alkaline-tolerant enzyme that is produced by Bacillus stearothermophilus T-6. Xylanase T-6 was found to bleach pulp effectively at pH 9 and 65 degrees C and was used successfully on an industrial-scale mill trial. To facilitate the future characterization of the protein via X-ray analysis and protein engineering, it was necessary to overexpress the enzyme in Escherichia coli. The xylanase gene was cloned into T-7 polymerase expression vectors and its expression was optimized. The enzyme was found to constitute over 70% of the cell protein and it was efficiently purified from the host proteins by a single heating step. Over 2 g soluble and active enzyme per 1 culture were achieved.
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