Cloning, functional expression and promoter analysis of xylanase III gene from Trichoderma reesei

Applied Microbiology and Biotechnology
W OgasawaraY Morikawa

Abstract

In this study, the xyn3 gene from the filamentous mesophilic fungus Trichoderma reesei (Hypocrea jecorina) PC-3-7 was cloned and sequenced. Analysis of the deduced amino acid sequence of XYN III revealed considerable homology with xylanases belonging to glycoside hydrolase family 10. These results show that XYN III is distinguishable from XYN I and XYN II, two other T. reesei xylanases that belong to the glycosidase family 11. When xyn3 was expressed in Escherichia coli, significant activity was observed in the cell-free extract, and higher activity (13.2 U/ml medium) was recovered from the inclusion bodies in the cell debris. The sequence of the 5'-upstream region of the gene in the parent strain QM9414 is identical to that of PC-3-7, although the expression level of xyn3 in PC-3-7 has been reported to be at least 1,000 times greater than in QM9414. These results suggest that xyn3 expression in T. reesei QM9414 is silenced. The consensus sequences for ACEI, ACEII, CREI, and the Hap2/3/5 protein complex are all present in the upstream region of xyn3. Deletion analysis of the upstream region revealed that two regions containing consensus sequences for the known regulatory elements play important roles for xyn3 expression.

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Citations

Jan 17, 2008·Applied Microbiology and Biotechnology·Yosuke ShidaYasushi Morikawa
Jan 17, 2009·Applied Microbiology and Biotechnology·Zinnia RahmanYasushi Morikawa
Jul 2, 2009·Applied Microbiology and Biotechnology·Sibtain AhmedAmer Jamil
Jan 30, 2008·Journal of Industrial Microbiology & Biotechnology·Ana Carolina Segato RizzattiMaria de Lourdes Teixeira de Moraes Polizeli
Dec 25, 2008·Biotechnology for Biofuels·Isabelle Herpoël-GimbertMarcel Asther
Jun 19, 2014·PloS One·Rafael Silva-RochaRoberto Nascimento Silva
Jan 12, 2013·Applied Microbiology and Biotechnology·Charles RutterRachel Chen
May 3, 2008·Fungal Genetics and Biology : FG & B·Takanori FurukawaYasushi Morikawa
Jul 22, 2014·The FEBS Journal·Peter BielyKristian B R Mørkeberg Krogh
Apr 15, 2016·Bioscience, Biotechnology, and Biochemistry·Yosuke ShidaWataru Ogasawara
May 4, 2016·Applied Microbiology and Biotechnology·Tomohiko MatsuzawaKatsuro Yaoi
Feb 9, 2018·Applied Microbiology and Biotechnology·Hiroki HirasawaWataru Ogasawara

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