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The glucose oxidase of Penicillium variabile P16: gene cloning, sequencing and expression
Author(s) -
Pulci V.,
D'Ovidio R.,
Petruccioli M.,
Federici F.
Publication year - 2004
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2004.01470.x
Subject(s) - biology , gene , southern blot , blot , heterologous expression , gene expression , glucose oxidase , microbiology and biotechnology , molecular cloning , dna–dna hybridization , genomic dna , heterologous , biochemistry , enzyme , recombinant dna
Aims: Isolation and characterization of the glucose oxidase (GOX)‐encoding gene from a Penicillium variabile strain (P16) having a high level of GOX activity and comparison of its expression with that of another strain of P. variabile (NRRL 1048) characterized by low GOX activity. Methods and Results: The gene, isolated by PCR consisted of 1818 bp encoding 605 amino acid residues. Gene expression was analysed by Northern blotting and compared with that of P. variabile NRRL 1048. The higher GOX activity of strain P16 appeared likely because of de novo mRNA synthesis. Southern blotting analyses of the genomic DNA showed that the hybridization pattern of the two strains differed for the size of hybridizing fragment detected by the probe and slightly for their signal intensity. Conclusions: The GOX‐encoding gene of P. variabile P16 was isolated and characterized to identify the molecular bases of its high level of expression and in view of improving enzyme production by developing a process based on heterologous expression. Significance and Impact of the Study: GOX‐encoding genes can be subjected to high difference in their expression levels. The P16 strain of P. variable producing large amount of GOX as well as its encoding gene might be exploited for industrial applications.