Journal of Microbiology

Hyangsoon Lim

2 Articles

Growth Inhibition of the Yeast Transformant by the Expression of a Chitinase from Coprinellus congregatus: Hyangsoon Lim , Hyoung T. Choi; J. Microbiol. 2010;48(5):706-708. Published online November 3, 2010; DOI: https://doi.org/10.1007/s12275-010-0272-x

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Coprinellus congregatus generates several chitinases during its entire life cycle: at the growing hyphal stage and at the mushroom autolysis stage. We have isolated a chitinase gene (chi1) from the mushroom tissue at the autolysing stage, and constructed a chitinase expression vector to get large amount of enzyme protein. Chitinase 1 (chi1) cDNA was heterologously expressed in Saccharomyces cerevisiae by gal1 promoter. The transformants showed no specific change in growth characteristics under normal growth conditions. However the expression of the gene by the gal1 promoter in the yeast transformants resulted in complete growth inhibition, while laccase expression by the gal1 promoter showed normal growth. The chitinase activities from the transformants were also more than 3 times higher than that of the recipient strain, and the chitinase expression by the real time-PCR also showed increased expression of the chi1 in the yeast transformant. Expression of a chitinase which was produced at the mushroom autolysing stage of C. congregatus resulted in yeast growth inhibition.

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Enhanced Expression of Chitinase during the Autolysis of Mushroom in Coprinellus congregatus: Hyangsoon Lim , Hyoung T. Choi; J. Microbiol. 2009;47(2):225-228. Published online May 2, 2009; DOI: https://doi.org/10.1007/s12275-008-0247-3

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Abstract PDF

Fungal cell walls consist of various glucans and chitin. An inky cap, Coprinellus congregatus, produced mushrooms at 25°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. congregatus 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.

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