Heterologous Expression Of Recombinant Cellulase In Escherichia Coli Rosetta (De3) And Bl21 (De3)

• Main Author: Casandra, Kana Brooke
• Format: Final Year Project Report
• Language: English; English
• Published: Universiti Malaysia Sarawak, (UNIMAS) 2018
• Subjects: Q Science (General)
• Online Access: http://ir.unimas.my/id/eprint/35379/1/Heterologous%20Expression%20Of%20Recombinant%20Cellulase%20In%20Escherichia%20Coli%20Rosetta%20%28De3%29%20And%20Bl21%20%28De3%29%2824pgs%29.pdf; http://ir.unimas.my/id/eprint/35379/4/Heterologous%20Expression..ft.pdf; http://ir.unimas.my/id/eprint/35379/

The heterologous expression of cellulase gene from Aspergillus sp. in E.coli strains of Rosetta (DE3) and BL2 l (DE3) has been studied to compare the recombinant protein production and to observe their activity, efficiency and production rate of recombinant cellulase. In previous study, an improved protein quality has been detected by using Rosetta, therefore further works need to be carried out to compare its protein production with BL2 l. The pFT AG-vector with the recombinant cellulase was used for expression studies and inserted into both Rosetta and BL2 l. Screening of cellullolyctic organism has failed to reveal the activity of recombinant cellulase as there is no growth detected on the media, as well as no formation of halo zone. Other than detection using the formation of halo zone, Bradford Assay and DNS assay was conducted to determine the cellulase activity of the bacterial in liquid medium. Recombinant cellulase from Rosetta was shown to have higher cellulase activity when the protein concentration was calculated using the standard graph. It was due to the fact that Rosetta has rare codons implemented inside them in order to yield higher protein production and better protein purity