Cloning and developmental expression of a metzincin family metalloprotease cDNA from oyster mushroomPleurotus ostreatus

Abstract

A cDNA clone, PoMTP, encoding a putative metzincin family metalloprotease was isolated from the expressed sequence tags of a basidiomycete Pleurotus ostreatus. The 5'-end sequence of PoMTP was determined by the 5'-RACE method. Full-length cDNA sequence (1140 bp) of PoMTP contained a 870 bp open reading frame encoding a protein product of 290 amino acids in addition to a 99 bp of 5'-untranslated sequence and a 171 bp of 3'-untranslated sequence with a poly(A) tail. The deduced amino-acid sequences of PoMTP contained an extensive zinc-binding consensus sequence and a so-called Met-turn sequence which are typical for the metzincin family of metalloproteases, indicating that the PoMTP protein belongs to the metzincin metalloproteases. Four cysteine residues were also observed in the zinc-binding region of PoMTP amino-acid sequence, which are known to be important for the structure and the function of some subfamilies of the metzincins. Comparison of the PoMTP in sequence database showed no significant homology with functionally known metalloproteases of Armillaria mellea, Grifola frondosa, Lentinula edodes, Pleurotus ostreatus, Schizophyllum commune and Tricholoma saponaceum in mushroom. Northern blot and qunatitative RT-PCR analyses indicated the PoMTP mRNA to be abundant at primordial and fruit body stages, but scarce at the mycelial stage, suggesting that the PoMTP metalloprotease plays an important role in mushroom fruiting.

Molecular Analysis of Ulilysin, the Structural Prototype of a New Family of Metzincin Metalloproteases

“…Another member studied in vitro is PoMTP metalloprotease from the edible oyster mushroom, P. ostreatus (SwissProt code Q5Y972). The mRNA of this 290-residue (pro)protein is abundant at primordial and fruit body stages, thus suggesting a role in mushroom fruiting (35). These authors proposed that this protein should be grouped with a series of putative fungal orthologues in a separate metzincin family termed eucolycins.…”

Genetic linkage map and expression analysis of genes expressed in the lamellae of the edible basidiomycete Pleurotus ostreatus

“…Other P. ostreatus genes diVerentially expressed in vegetative and reproductive mycelia have been previously reported Joh et al, 2004;Peñas et al, 1998Peñas et al, , 2005. The largest study of diVerentially expressed genes on this species has been reported by Lee et al (2002), which compared ESTs isolated from liquid cultures and fruit bodies.…”

Purification and Characterization of a Fibrinolytic Protease from a Culture Supernatant ofFlammulina velutipesMycelia

“…[16][17][18] These constitute an important source of thrombolytic agents. Some reports have described the fibrinolytic activity of certain edible mushrooms, including Armillaria mellea, 19,20) Grifora frondosa, 21) Pleurotus ostreatus, 22,23) and Cordyceps militaris. 24) The golden needle mushroom, Flammulina velutipes, belonging to the family Tricolomataceae (Hymenomycetes, Basidiomycota), is quite popular in Korea, Japan, China, and Taiwan.…”