Cloning and heterologous expression of the exo-beta-D-glucosaminidase-encoding gene (gls93) from a filamentous fungus, Trichoderma reesei PC-3-7

Applied Microbiology and Biotechnology
Masakazu IkeYasushi Morikawa

Abstract

We have previously reported on purification and characterization of an exo-beta-D-glucosaminidase (Gls93) from culture filtrate of Trichoderma reesei PC-3-7 grown on N-acetyl-D-glucosamine (GlcNAc). The corresponding gene of Gls93 was cloned and characterized in this work. To our knowledge, this is the first report on cloning of the gene encoding fungal exo-beta-D-glucosaminidase. This gene has no introns and encodes a polypeptide of 892 amino acids (aa) containing a secretion signal of 28 amino acids. Comparison of the amino acid sequence to known proteins and phylogenetic analysis indicated that gls93 belongs to the glycoside hydrolase family (GHF) 2 and should be further classified into a new subgroup, exo-beta-D-glucosaminidase subgroup. The gls93 transcription was biphasic when T. reesei was grown on GlcNAc, suggesting that the expression of this gene may be regulated by a complex mechanism, in which multiple regulatory proteins are involved. Furthermore, gls93 could be expressed in Pichia pastoris (ca. 0.49-mg/ml culture). The recombinant Gls93 had the two molecular forms, ca. 105 and 100 kDa, whose difference is caused by N-glycosylation. Both of them had the same properties such as specific activity and substrate specif...Continue Reading

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Citations

Dec 3, 2011·Applied Microbiology and Biotechnology·Lukas HartlVerena Seidl-Seiboth
Mar 9, 2010·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Yasumitsu SakamotoKazuo T Nakamura
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Mar 27, 2012·Fungal Genetics and Biology : FG & B·Erzsébet FeketeMichel Flipphi
Apr 23, 2013·Biotechnology Progress·S B Chavan, M V Deshpande
Aug 11, 2020·Fungal Genetics and Biology : FG & B·Yunhan Hsu, Manabu Arioka
Jan 5, 2021·Journal of Agricultural and Food Chemistry·Bhoopal BhuvanachandraAppa Rao Podile

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