Expression of Trichoderma reesei endo-beta-glucanase II in silkworm, Bombyx mori L. by using BmNPV/Bac-to-Bac expression system and its bioactivity assay.
The silkworm, Bombyx mori, was used to produce recombinant endo-beta-glucanase II (rEGII). The EGII gene (egl2) was cloned from the cellulolytic fungus Trichoderma reesei and inserted into B. mori nucleopolyhedrovirus (BmNPV) genome using BmNPV/Bac-to-Bac expression vector. For expression of rEGII, both the BmN cells and B. mori larvae were infected with the recombinant virus. The putative rEGII yield was about 386 microg per larva and the enzyme activity of the purified rEGII was approx 352 U/mg of rEGII. The optimal activity of this purified protein was observed at 55 degrees C and pH 4, respectively.
Zhou
Lie
L
Institute of Dairy Science, Ministry of Education Key Laboratory of Molecular Animal Nutrition, Zhejiang University, Hangzhou, 310029, People's Republic of China.
Wu
Xiaofeng
X
Lan
Lipan
L
Liu
Jianxin
J
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D012831:Bombyx / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002480:Cellulase / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D007814:Larva / Q000235:genetics / Q000378:metabolism; D014242:Trichoderma / Q000201:enzymology* / Q000235:genetics
D005656:Fungal Proteins; D002480:Cellulase