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Expression and Enzyme Activity Determination of Apripona Germari Cellulase in Bmnpv/Bac-to-Bac Baculovirus Expression Systems

Lu Gao1, Zhe Yang2, Keping Chen2

Abstract

To Search for a more efficient way of degrading cellulose. METHODS: Apripona germari Cellulase genes were cloned and expressed in Bombyx mori derived cell line, BmN cell line by BmNPV/Bac-to-Bac baculovirus expression systems. Infected BmN cells were harvested 72 h post-infection (hpi). Expressd recombinant cellulose was analyzed in 12.5% SDS– PAGE and western blot. The activity of purified recombinant cellulose was assessed by 3, 5- nitro salicylic acid (DNS) colorimetric method. RESULTS: Western blot analysis showed that Recombinant cellulase was expressed in BmN insect cells as a 29 kDa protein. Enzyme activity assay demonstrated that recombinant cellulose has the activity of decomposing cellulose. CONCLUSION: Apripona germari Cellulase was successfully expressed in BmN insect cell line. The method established in this study provides an efficient way to produce a large amount of cellulase and paves the way for further utilization of cellulose.


Keywords

Apripona Germari; Cellulase; Bac-to-Bac Baculovirus Expressio; Enzyme Activity

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DOI: http://dx.doi.org/10.18063/bc.v2i1.570
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