Cloning, Sequencing, and Expression of a Thermostable Cellulase Gene ofHumicola grisea

Structure and Characteristics of an Endo-β-1,4-glucanase, Isolated fromTrametes hirsuta, with High Degradation to Crystalline Cellulose

“…The cDNA encoded 384 amino acids with a calculated M r of 40.6 k. As shown in Fig. 3, a comparison of amino acid sequences in the BLAST search (only of those whose gene was registered) revealed high homology with the sequences of Irpex lacteus En-1 (73% identity, AB194135), 10) Humicola grisea EG (46%, D84470), 22) and A. niger EglB (49%, AJ224452). 23) These are all endo--1,4-glucanases belonging to GH family 5 16) containing family 1 CBM, except for A. niger EglB.…”

“…16) These fungal enzymes contain CBM in the N-terminal. 10,22) On this basis, the 44-k protein was postulated to be an endo--1,4-glucanase belonging to GH family 5. It was named ThEG.…”

Cloning and expression of endo-β-glucanase II gene in Humicola insolens H31-3

“…3) and sequenced. Sequence alignment with other cellulase gene sequence indicated that egl2 cDNA sequence had a high similarity to that of H. grisea egl2 (Takashima et al, 1997), about 98.9% homogeneity, and H. insolens CMC3 (Dalboge and HeldtHansen, 1994), about 97.26% homogeneity.…”

“…Primers P1 and P2, designed according to the reported sequence (Takashima, 1997;Dalboge and Heldt-Hansen, 1994), were applied to amplify the sequence of egl2 cDNA (P 1 :59-ATCTCGAGCAGGGCGGTGCATGG-39, P 2 :59-TAGCGGCCGCCTATGGCACGTATT-39). PCR was performed using TaKaRa LA Taq H .…”

“…A number of cellulase genes has been cloned, sequenced, and expressed (Dalboge and Heldt-Hansen, 1994;Takashima et al, 1997;Moriya et al, 2003). Yet, only very low yields of enzyme are obtained from culture fluids using conventional purification techniques, even when the fungus is grown under conditions optimal for enzyme biosynthesis.…”