Open AccessThe nagA gene of Penicillium chrysogenum encoding β‐ N ‐acetylglucosaminidase
Author(s) -
Díez Bruno,
RodríguezSáiz Marta,
La Fuente Juan Luis,
Moreno Miguel Ángel,
Barredo José Luis
Publication year - 2005
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/j.femsle.2004.11.017
Subject(s) - penicillium chrysogenum , gene , genetics , biology , microbiology and biotechnology , chemistry
We purified the β‐ N ‐acetylglucosaminidase from the filamentous fungus Penicillium chrysogenum and its N‐terminal sequence was determined, showing the presence of a mixture of two proteins (P1 and P2). A genomic DNA fragment was cloned by using degenerated oligonucleotides from the Nt sequences. The nucleotide sequence showed the presence of an ORF ( nagA gene) lacking introns, with a length of 1791 bp, and coding for a protein of 66.5 kDa showing similarity to acetylglucosaminidases. The NagA deduced protein includes P1 and P2 as incomplete forms of the mature protein, and contains putative features for protein maturation: an 18‐amino acid signal peptide, a KEX2 processing site, and four glycosylation motifs. The sequence just after the signal peptide corresponds to P2 and that after the KEX2 site to P1. The nagA transcript has a size of about 2.1 kb and is present until the end of the fermentation process for penicillin production. NagA is one of the most largely represented proteins in P. chrysogenum , increasing along the fermentation process. The suitability of the nagA promoter (P nagA ) for gene expression in fungi was demonstrated by expressing the bleomycin resistance gene ( ble R ) from Streptoalloteichus hindustanus in P. chrysogenum .