Heterologous Expression Characteristics of Trichoderma viride Endoglucanase V in the Silkworm, Bombyx mori L.

Li, Xing-hua; Wang, Mei-xian; Zhang, Peng; Hu, Jia-biao; Sun, Chun-guang; Liu, Xin-ju; Zhou, Fang; Niu, Yan-shan; Malik, Firdose; Bhaskar, Roy; Yang, Hua-jun; Miao, Yun-gen

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.

ENZYMES; GLUCOSIDASES; TRICHODERMA; SILKWORMS; NUCLEOPOLYHEDROVIRUSES; CELLULASE

Applied Biochemistry & Biotechnology, 2011, Vol 165, Issue 2, p728

0273-2289

Academic Journal

10.1007/s12010-011-9291-2