A novel thermostable xylanase GH10 from Malbranchea pulchella expressed in Aspergillus nidulans with potential applications in biotechnology

Biotechnology for Biofuels

Liliane FC Ribeiro³  Rosymar C De Lucas³  Gabriela L Vitcosque³  Lucas F Ribeiro³  Richard J Ward⁵  Marcelo V Rubio⁵  Andre RL Damásio⁵  Fabio M Squina⁵  Rebecca C Gregory⁶  Paul H Walton⁶  João A Jorge²  Rolf A Prade¹  Marcos S Buckeridge⁴  Maria de Lourdes TM Polizeli² 

[1] Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK, USA

[2] Biology Department of Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto - USP, Av. Bandeirantes, 3900, Ribeirão Preto, SP 14040-901, Brazil

[3] Immunology and Biochemistry Department of Faculdade de Medicina de Ribeirão Preto - USP, Ribeirão Preto, SP, Brazil

[4] Institute of Biosciences, University of São Paulo, São Paulo, Brazil

[5] Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Campinas, SP, Brazil

[6] Department of Chemistry, The University of York, York, UK

关键词: Sugarcane bagasse;    Biomass degradation;    Thermostability;    Heterologous expression;    Glycosylation;    Malbranchea;    Xylanase;   

Others  :  1084764
DOI  :  10.1186/1754-6834-7-115

 received in 2014-02-28, accepted in 2014-07-15,  发布年份 2014

【 摘 要 】

Background

The search for novel thermostable xylanases for industrial use has intensified in recent years, and thermophilic fungi are a promising source of useful enzymes. The present work reports the heterologous expression and biochemical characterization of a novel thermostable xylanase (GH10) from the thermophilic fungus Malbranchea pulchella, the influence of glycosylation on its stability, and a potential application in sugarcane bagasse hydrolysis.

Results

Xylanase MpXyn10A was overexpressed in Aspergillus nidulans and was active against birchwood xylan, presenting an optimum activity at pH 5.8 and 80°C. MpXyn10A was 16% glycosylated and thermostable, preserving 85% activity after 24 hours at 65°C, and deglycosylation did not affect thermostability. Circular dichroism confirmed the high alpha-helical content consistent with the canonical GH10 family (β/α)₈ barrel fold observed in molecular modeling. Primary structure analysis revealed the existence of eight cysteine residues which could be involved in four disulfide bonds, and this could explain the high thermostability of this enzyme even in the deglycosylated form. MpXyn10A showed promising results in biomass degradation, increasing the amount of reducing sugars in bagasse in natura and in three pretreated sugarcane bagasses.

Conclusions

MpXyn10A was successfully secreted in Aspergillus nidulans, and a potential use for sugarcane bagasse biomass degradation was demonstrated.

【 授权许可】

   
2014 Ribeiro et al.; licensee BioMed Central Ltd.

【 图 表 】

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