Molecular cloning and sequences of lignin peroxidase genes of Phanerochaete chrysosporium.

Abstract

The genomic clones encoding lignin peroxidase isozyme H8 and two closely related genes were isolated from Phanerochaete chrysosporium BKM-1767, and their nucleotide sequences were determined. The positions and approximate lengths of introns were found to be highly conserved in all three clones. Analysis of homokaryotic derivatives indicated that the three clones are not alleles of the same gene(s).

Lignin peroxidase gene family of Phanerochaete chrysosporium: complex regulation by carbon and nitrogen limitation and identification of a second dimorphic chromosome

“…Deduced amino acid sequences exceed 80% identity in any pairwise comparison within this subfamily. In contrast, LiP genes CLG4 (7) and V4 (30) are distinctly different from each other and from members of the subfamily.…”

“…Cosmid pQQ24 is a pWE15 derivative (35) containing LiP genes LiPA, LiPB, and GLG5 (9). Single LiP genes are contained in cosmid pKBY2 (38) derivatives and in pO282 and pV4 (30). Subclone pO282::910 contains a 910-bp XhoI fragment of 0282 in Bluescript KSII (Stratagene Inc., La Jolla, Calif.).…”

“…Oligonucleotide 5'-GTACGTGGTCTCGATCGAGG-3' corresponds to the reverse complement of CLG4 gene (7) bp 343 to 362, a region of little homology to related LiP genes. Oligonucleotide 5'-GAAATTGGTCTCGATAGTAT-3' is V4 specific, lacking significant homology with all LiP genes except the reverse complement of the V4 gene at a position 37 bp downstream of the third intron (30). Probes were synthesized by the ,-cyanoethyl phosphoramidite method on an Applied Biosystems (Foster City, Calif.) model 391 without further purification and then 32P end labeled with T4 polynucleotide kinase (28).…”

“…In the widely used strain BKM-1767, six genes plus allelic variants have been identified (7,9,16,30,31,33). Four LiP genes (LiPA, LiPB, GLG5, and 0282) are closely related with respect to nucleotide sequence, intron positions, and codon preference.…”

A novel extrachromosomally maintained transformation vector for the lignin-degrading basidiomycete Phanerochaete chrysosporium

“…Genetic studies with this organism have been limited, but auxotrophic mutants (15,26,27) and mutants lacking various secondary metabolic activities (7,21,24) have been isolated. Both cDNA (13,49,53,55) and genomic clones (3,5,8,20,41,42,52,54) for lignin peroxidases and cDNA clones for manganese peroxidases (31,34) have been isolated and sequenced. We previously described vector pRR12, which was shown to transform P. chrysosporium to G418 resistance and was rescued in Escherichia coli from the total DNA of pRR12 transformants (35).…”