Expression of <Emphasis Type="Italic">Trichoderma</Emphasis><Emphasis Type="Italic">viride</Emphasis> endoglucanase III in the larvae of silkworm, <Emphasis Type="Italic">Bombyx mori L.</Emphasis> and characteristic analysis of the recombinant protein |
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Authors: | Xing-hua Li Peng Zhang Mei-xian Wang Fang Zhou Firdose Ahmad Malik Hua-jun Yang Roy Bhaskar Jia-biao Hu Chun-guang Sun Yun-gen Miao |
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Institution: | (1) Key Laboratory of Animal Epidemic Etiology and Immunological Prevention of Ministry of Agriculture, Zhejiang University, Hangzhou, 310029, People’s Republic of China;(2) College of Animal Sciences, Zhejiang University, Hangzhou, 310029, People’s Republic of China |
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Abstract: | Endoglucanase is a part of cellulase which hydrolyzes cellulose into glucose. In this study, we cloned endoglucanase III (EG
III) gene from Trichoderma viride strain AS 3.3711 using a PCR-based exon splicing method, and expressed EG III recombinant protein in both silkworm BmN cell
line and silkworm larvae with an improved Bac-to-Bac/BmNPV mutant baculovirus expression system, which lacks the chiA and v-cath genes of Bombyx mori nucleopolyhedrovirus (BmNPV). The result showed that around 45 kDa protein was visualized in BmN cells at 48 h after the
second generation recombinant mBacmid/BmNPV/EG III baculovirus infection. The enzymes from recombinant baculoviruses infected
silkworms exhibited significant maximum enzyme activity at the environmental condition of pH 8.0 and temperature 50°C, and
increased 20.94 and 19.13% compared with that from blank mBacmid/BmNPV baculoviruses infected silkworms and normal silkworms,
respectively. It was stable at pH range from 5.0 to 9.0 and at temperature range from 40 to 60°C. It provided a possibility
to generate transgenic silkworms expressing bio-active cellulase, which can catabolize dietary fibers more efficiently, and
it might be of great significance for sericulture industry. |
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