2014•Mycologia iranicaOpen access

Expression analyses of endoglucanase gene in Penicillium oxalicum and Trichoderma viride

S. Leylaie, Seyed Kazem Sabbagh, M. A. Tajick Ghanbari

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Abstract

The expression of endoglucanase gene and protein profile belonging to two fungal species, Penicillium oxalicum 1SMS and Trichoderma viride 156MS with high cellulase enzyme activity, was investigated. Fungal isolates were cultured on inducer CMC medium and then the amount of released sugar and protein were assayed every three days for a month, using arsenate molybdate reagent and Bradford method, respectively. Detection of endoglucanase gene was performed by using specific primers. Expressed cDNA fragments in two isolates revealed a size of 1380 bp to 1434 bp length. The cDNAs were sequenced and queried in BLASTN and TBLASTX search against eg1 genes from other fungi. These sequences showed 98% homology to eg1 gene deposited in databases. SDS-PAGE pattern of protein separation revealed one protein band for each isolate having a molecular weight of 49.786 and 48.339 kDa with 487 and 460 amino acids. Bioinformatics analysis revealed glycosyl hydrolase domains, a cellulase banding domain and 6 PROSITEs for both proteins. One ORF in each gene region was determined for each species. The results indicated that carboxy methyl cellulose is able to induce expression of eg1, following endogluconase enzyme production.

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What this paper is about

The expression of endoglucanase gene and protein profile belonging to two fungal species, Penicillium oxalicum 1SMS and Trichoderma viride 156MS with high cellulase enzyme activity, was investigated. Fungal isolates were cultured on inducer CMC medium and then the amount of released sugar and protein were assayed every three days for a month, using arsenate molybdate reagent and Bradford method, respectively. Detection of endoglucanase gene was performed by using specific primers. Expressed cDNA fragments in two isolates revealed a size of 1380 bp to 1434 bp length. The cDNAs were sequenced and queried in BLASTN and TBLASTX search against eg1 genes from other fungi. These sequences showed 98% homology to eg1 gene deposited in databases. SDS-PAGE pattern of protein separation revealed one protein band for each isolate having a molecular weight of 49.786 and 48.339 kDa with 487 and 460 amino acids. Bioinformatics analysis revealed glycosyl hydrolase domains, a cellulase banding domain and 6 PROSITEs for both proteins. One ORF in each gene region was determined for each species. The results indicated that carboxy methyl cellulose is able to induce expression of eg1, following endogluconase enzyme production.

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Available abstract

The expression of endoglucanase gene and protein profile belonging to two fungal species, Penicillium oxalicum 1SMS and Trichoderma viride 156MS with high cellulase enzyme activity, was investigated. Fungal isolates were cultured on inducer CMC medium and then the amount of released sugar and protein were assayed every three days for a month, using arsenate molybdate reagent and Bradford method, respectively. Detection of endoglucanase gene was performed by using specific primers. Expressed cDNA fragments in two isolates revealed a size of 1380 bp to 1434 bp length. The cDNAs were sequenced and queried in BLASTN and TBLASTX search against eg1 genes from other fungi. These sequences showed 98% homology to eg1 gene deposited in databases. SDS-PAGE pattern of protein separation revealed one protein band for each isolate having a molecular weight of 49.786 and 48.339 kDa with 487 and 460 amino acids. Bioinformatics analysis revealed glycosyl hydrolase domains, a cellulase banding domain and 6 PROSITEs for both proteins. One ORF in each gene region was determined for each species. The results indicated that carboxy methyl cellulose is able to induce expression of eg1, following endogluconase enzyme production.

Key concepts: Trichoderma viride, Cellulase, Biology, Gene, Complementary DNA, Gene expression, Trichoderma, Biochemistry

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Expression analyses of endoglucanase gene in Penicillium oxalicum and Trichoderma viride — Research Paper | ScholarLens