Abstract
A xylanase gene from Paecilomyces thermophila was functionally expressed in Pichia pastoris. The recombinant xylanase (xynA) was predominantly extracellular; in a 5 l fermentor culture, the total extracellular protein was 8.1 g l−1 with an activity of 52,940 U ml−1. The enzyme was purified to homogeneity with a recovery of 48 %. The recombinant xynA was optimally active at 75 °C, as measured over 10 min, and at pH 7. The enzyme was stable up to 80 °C for 30 min. It hydrolyzed birchwood xylan, beechwood xylan and xylooligosaccharides to produce xylobiose and xylotriose as the main products.
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References
Bakalova NG, Petrova SD, Kolev DN (2002) Biochemical and catalytic properties of endo-1,4-β-xylanases from Thermomyces lanuginosus (wild and mutant strains). Biotechnol Lett 24:1167–1172
Chadha BS, Ajay BK, Mellon F, Bhat MK (2004) Two endoxylanases active and stable at alkaline pH from the newly isolated thermophilic fungus Myceliophthora sp. IMI 387099. J Biotechnol 109:227–237
Damaso MC, Almeida MS, Kurtenbach E, Martins OB, Pereira N, Andrade CM, Albano RM (2003) Optimized expression of a thermostable xylanase from Thermomyces lanuginosus in Pichia pastoris. Appl Environ Microbiol 69:6064–6072
Fawzi EM (2010) Highly thermostable purified xylanase from Rhizomucor miehei NRRL 3169. Ann Microbiol 60:363–368
Gaffney M, Garberry S, Doyle S, Murphy R (2009) Purification and characterization of a xylanase from Thermomyces lanuginosus and its functional expression by Pichia pastoris. Enzyme Microb Technol 45:348–354
He J, Yu B, Zhang K, Ding X, Chen D (2009) Expression of endo-1,4-β-xylanase from Trichoderma reesei in Pichia pastoris and functional characterization of the produced enzyme. BMC Biotechnol 9:56–66
Katrolia P, Zhou P, Zhang P, Yan QJ, Li YN, Jiang ZQ, Xu HB (2012) High level expression of a novel β-mannanase from Chaetomium sp. exhibiting efficient mannan hydrolysis. Carbohydr Polym 87:480–490
Lin J, Ndlovu LM, Singh S, Pillay B (1999) Purification and biochemical characteristics of β-d-xylanase from a thermophilic fungus, Thermomyces lanuginosus-SSBP. Biotechnol Appl Biochem 30:73–79
Luo HY, Li J, Yang J, Wang H, Yang YH, Huang HQ, Shi PJ (2009) A thermophilic and acid stable family-10 xylanase from the acidophilic fungus Bispora sp. MEY-1. Extremophiles 13:849–857
Maalej I, Belhaj I, Masmoudi NF, Belghith H (2009) Highly thermostable xylanase of the thermophilic fungus Talaromyces thermophilus: purification and characterization. Appl Biochem Biotechnol 158:200–212
Patrick S, Fazenda ML, McNeil B, Harvey LM (2005) Heterologous protein production using the Pichia pastoris expression system. Yeast 22:249–270
Polizeli MLTM, Rizzatti ACS, Monti R, Terenzi HF, Jorge JA, Amorim DS (2005) Xylanases from fungi: properties and industrial applications. Appl Microbiol Biotechnol 67:577–591
Sriyapai T, Somyoonsap P, Matsui K, Kawai F, Chansiri K (2011) Cloning of a thermostable xylanase from Actinomadura sp. S14 and its expression in Escherichia coli and Pichia pastoris. J Biosci Bioeng 111:528–536
Yang SQ, Yan QJ, Jiang ZQ, Li LT, Tian HM, Wang YZ (2006) High-level of xylanase production by the thermophilic Paecilomyces themophila J18 on wheat straw in solid-state fermentation. Bioresour Technol 97:1794–1800
Zhang M, Jiang ZQ, Yang SQ, Hua CW, Li LT (2010) Cloning and expression of a Paecilomyces thermophila xylanase gene in E. coli and characterization of the recombinant xylanase. Bioresour Technol 101:688–695
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This work was financially supported by the National High Technology Research and Development Program of China (863 Program, No. 2011AA100905).
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Fan, G., Katrolia, P., Jia, H. et al. High-level expression of a xylanase gene from the thermophilic fungus Paecilomyces thermophila in Pichia pastoris . Biotechnol Lett 34, 2043–2048 (2012). https://doi.org/10.1007/s10529-012-0995-3
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DOI: https://doi.org/10.1007/s10529-012-0995-3