Growth inhibition of the yeast transformant by the expression of a chitinase from Coprinellus congregatus.
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.
Department of Biochemistry, Kangwon National University, Chunchon 200-701, Republic of Korea.
Hyangsoon
H
Lim
Hyoung T
HT
Choi
D000363:Agaricales / Q000201:enzymology* / Q000235:genetics; D002688:Chitinases / Q000096:biosynthesis* / Q000235:genetics / Q000633:toxicity*; D003001:Cloning, Molecular; D015870:Gene Expression; D020869:Gene Expression Profiling; D006131:Growth Inhibitors / Q000096:biosynthesis* / Q000235:genetics / Q000633:toxicity*; D042845:Laccase / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000633:toxicity; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development*; D014170:Transformation, Genetic
D006131:Growth Inhibitors; D012333:RNA, Messenger; D011994:Recombinant Proteins; D042845:Laccase; D002688:Chitinases