title: Heterologous expression characteristics of Trichoderma viride endoglucanase V in the silkworm, Bombyx mori L.
abstract: Efficient degradation of cellulose needs a synergistic reaction of the cellulolytic enzymes, which include exoglucanases, endoglucanases, and β-1,4-glucosidase. In this study, we used an improved Bac-to-Bac/BmNPV baculovirus expression system, which lacks the virus-encoded chitinase cathepsin (v-cath) genes of Bombyx mori nucleopolyhedrovirus (BmNPV), to express the endoglucanase V (EG V) gene from Trichoderma viride in silkworm BmN cells and silkworm larvae, and analyzed the characteristics of the recombinant enzyme in silkworm larvae. The result showed that an around 36-kDa protein was visualized in BmN cells at 48 h after the second-generation recombinant mBacmid/BmNPV/EG V baculovirus infection. The crude enzyme extract from the recombinant baculoviruses-infected silkworms exhibited a significant maximum activity at the environmental condition of pH 5.0 and a temperature of 50 °C, and increased 39.86% and 37.76% compared with that from blank mBacmid/BmNPV baculovirus-infected silkworms and normal silkworms, respectively. It was stable at pH range from 5.0 to 10.0 and at temperature range from 40 to 60 °C. The availability of large quantities of EG V that the silkworm provides might greatly facilitate the future research and the potential application in industries.
authors.affiliation: Key Laboratory of Animal Epidemic Etiology and Immunological Prevention of Ministry of Agriculture, Zhejiang University, Hangzhou 310029, People's Republic of China.
authors.forename: Xing-Hua
authors.initials: XH
authors.lastname: Li
authors.forename: Mei-Xian
authors.initials: MX
authors.lastname: Wang
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.forename: Jia-Biao
authors.initials: JB
authors.lastname: Hu
authors.forename: Chun-Guang
authors.initials: CG
authors.lastname: Sun
authors.forename: Xin-Ju
authors.initials: XJ
authors.lastname: Liu
authors.forename: Fang
authors.initials: F
authors.lastname: Zhou
authors.forename: Yan-Shan
authors.initials: YS
authors.lastname: Niu
authors.forename: Firdose Ahmad
authors.initials: FA
authors.lastname: Malik
authors.forename: Roy
authors.initials: R
authors.lastname: Bhaskar
authors.forename: Hua-Jun
authors.initials: HJ
authors.lastname: Yang
authors.forename: Yun-Gen
authors.initials: YG
authors.lastname: Miao
mesh_terms: D000818:Animals; D001673:Biodegradation, Environmental; D001709:Biotechnology / Q000379:methods*; D015153:Blotting, Western; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D002403:Cathepsins / Q000172:deficiency / Q000235:genetics; D002460:Cell Line; D002480:Cellulase / Q000096:biosynthesis* / Q000235:genetics; D002482:Cellulose / Q000378:metabolism*; D002688:Chitinases / Q000172:deficiency / Q000235:genetics; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D007814:Larva / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D017924:Nucleopolyhedroviruses / Q000201:enzymology / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D014242:Trichoderma / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D002482:Cellulose; C097231:endoglucanase V; D002688:Chitinases; D002480:Cellulase; D002403:Cathepsins