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

Content Provider	Springer Nature Link
Author	Ike, Masakazu Isami, Koji Tanabe, Yoshio gawa, Masahiro Ogasawara, Wataru Okada, Hirofumi Morikawa, Yasushi
Copyright Year	2006
Abstract	We have previously reported on purification and characterization of an exo-β-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-β-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-β-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 specificity and showed only the activity of exo-β-d-glucosaminidase but not those of β-galactosidase, β-glucuronidase, and β-mannosidase belonging to GHF2.
Starting Page	687
Ending Page	695
Page Count	9
File Format	PDF
ISSN	01757598
Journal	Applied Microbiology and Biotechnology
Volume Number	72
Issue Number	4
e-ISSN	14320614
Language	English
Publisher	Springer-Verlag
Publisher Date	2006-04-25
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Microbial Genetics and Genomics Microbiology Biotechnology
Content Type	Text
Resource Type	Article
Subject	Medicine Applied Microbiology and Biotechnology Biotechnology

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