Enhanced expression of chitinase during the autolysis of mushroom in Coprinellus congregatus

Abstract

Fungal cell walls consist of various glucans and chitin. An inky cap, Coprinellus congregates, produced mushrooms at 25 degrees C in a regime of 15 h light/9 h dark, and then the mushroom was autolyzed rapidly to generate black liquid droplets where no cell wall was detected by microscopy. A chitinase cDNA from the matured mushroom cells of C. congregates that consisted of 1,541 nucleotides was successfully cloned using the rapid amplification of cDNA ends (RACE)-PCR technique. Its deduced 441 amino acid sequence had the conserved catalytic domain as in other fungal chitinase family 18. Chitinase activity was higher at the matured mushroom stage than primordial and young mushroom stage. When the expression of the cloned chitinase was examined by real-time PCR using the chitinase-specific primers, it was increased more than twice to 20 times during the autolytic process of mushroom than young mushroom or primordial stages, respectively.

Purification, characterization and synergism in autolysis of a group of 1,3-β-glucan hydrolases from the pilei of Coprinopsis cinerea fruiting bodies

“…The activities of these glycoside hydrolases have been detected in the extracts and autolysates of fruiting bodies of some species of Coprinopsis (Bush, 1974;Iten & Matile, 1970;Kamada et al, 1985;Lim & Choi, 2009). However, successful isolation, purification and characterization of these different hydrolases from Coprinopsis mycelium or fruiting bodies have not been reported to date.…”

“…During the development of fruiting bodies, basidiomycetes produce some hydrolytic enzymes associated with these polysaccharides in cell walls. These fungal glycoside hydrolases act on cell wall components and are considered to be responsible for cell wall extension (Kamada et al, 1991;Lim & Choi, 2009;Muraguchi et al, 2008;Tao et al, 2013), cell wall remodelling (Bowman & Free, 2006;Cabib & Arroyo, 2013;Gastebois et al, 2009;Mouyna et al, 2013) and fruiting body autolysis (Fukuda et al, 2008;Kües, 2000;Minato et al, 2004;Sakamoto et al, 2012;Tao et al, 2013).…”

Growth inhibition of the yeast transformant by the expression of a chitinase from Coprinellus congregatus

“…When the autolyzed tissue and the liquid droplet were examined by the light and electron microscopy, the cell walls of basidia disintegrated during autolysis and no cell wall was observed in the liquid droplet (Choi and Cho, 2005). A chitinase cDNA and genomic DNA were isolated from the cells of autolysed mushroom tissues, and reported that the cloned chitinase gene (chi1) was most highly expressed at the mushroom maturation/autolysis stage (Lim and Choi, 2009). We would like to analyze biochemical characteristics of the purified chitinase, but it was too hard to get large amount of the autolysing mushroom tissue under lab conditions.…”

Peer Review #3 of "Postharvest biochemical characteristics and ultrastructure of Coprinus comatus (v0.1)"

“…Studies have addressed that the content of chitin decreased during postharvest storage due to the effect of chitinase by hydrolyzing the β-1, 4-glycoside bond in chitin and producing n-acetylglucosamine oligomer or monomer (Buitimea et al, 2013;. Lim & Choi (2009) revealed that the Agaricus bisporus could rapidly produce black juice at 25°C for 15h after harvest and confirmed that the amount of chitinase synthesis in the autolysis process was significantly higher than that of unpicked mushrooms. Xue (2012) studied the effect of postharvest calcium treatment on cell stability of Lentinula edodes and found that the stability of cell wall was correlated with β-1, 3-glucan and β-1, 3-glucanase, by inhibiting the activity of β-1, 3-glucanase, the aging of L. edodes could be delayed.…”