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Heterologous expression of β-xylosidase gene from Paecilomyces thermophila in Pichia pastoris

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β-xylosidase from thermophilic fungi Paecilomyces thermophila was functionally expressed in Pichia pastoris with a his tag in the C-terminal under the alcohol oxidase 1 (AOX1) promoter and secreted into the medium at 0.22 mg l−1. Its molecular mass was estimated to be 52.3 kDa based on the SDS-PAGE analysis, which is 1.3 times higher than the predicted 39.31 kDa from its amino acid compositions, although no potential N- or O- glycosylation sites were predicted from its amino acid sequence. This is presumed to be caused by some unpredictable posttranslational modifications based on mass spectrum analysis of the recombinant protein. The enzyme was most active at 60 °C and pH 7. It showed not only a β-xylosidase activity with a Km of 8 mM and a Vmax of 54 μmol min−1 mg−1 for hydrolysis of p-nitrophenyl β-d-xylopyranoside but also an arabinofuranosidase activity (6.2 U mg−1) on p-nitrophenyl arabinofuranoside.

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This research is supported by the Science and Engineering Research Council (SERC) of the Agency for Science, Technology and Research (A*STAR) of Singapore (SERC grant no 0921590132). Thanks are given to Dr. Manfred Raida at Biopolis Shared Facility of A*STAR for help in mass spectrum analysis.

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Correspondence to Jin Chuan Wu.

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Juturu, V., Wu, J.C. Heterologous expression of β-xylosidase gene from Paecilomyces thermophila in Pichia pastoris . World J Microbiol Biotechnol 29, 249–255 (2013).

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