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. 2016 Jun;6(6):285.
doi: 10.4172/2155-9821.1000285. Epub 2016 Jun 30.

Synthesizing a Cellulase like Chimeric Protein by Recombinant Molecular Biology Techniques

Hirendra Nath Banerjee  1 , Christopher Krauss  1 , Valerie Smith  1 , Kelly Mahaffey  1 , Ava Boston  1
Affiliations

Synthesizing a Cellulase like Chimeric Protein by Recombinant Molecular Biology Techniques

Hirendra Nath Banerjee et al. J Bioprocess Biotech. 2016 Jun.

Abstract

In order to meet the Renewable Fuels Standard demands for 30 billion gallons of biofuels by the end of 2020, new technologies for generation of cellulosic ethanol must be exploited. Breaking down cellulose by cellulase enzyme is very important for this purpose but this is not thermostable and degrades at higher temperatures in bioreactors. Towards creation of a more ecologically friendly method of rendering bioethanol from cellulosic waste, we attempted to produce recombinant higher temperature resistant cellulases for use in bioreactors. The project involved molecular cloning of genes for cellulose-degrading enzymes based on bacterial source, expressing the recombinant proteins in E. coli and optimizing enzymatic activity. We were able to generate in vitro bacterial expression systems to produce recombinant His-tag purified protein which showed cellulase like activity.

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Figures

Figure 1
Figure 1
Nucleotide sequence of the PCR amplified amplicon.
Figure 2
Figure 2
NCBI-BLAST search result of the sequenced amplicon DNA.
Figure 2
Figure 2
NCBI-BLAST search result of the sequenced amplicon DNA.
Figure 2
Figure 2
NCBI-BLAST search result of the sequenced amplicon DNA.
Figure 2
Figure 2
NCBI-BLAST search result of the sequenced amplicon DNA.
Figure 3
Figure 3
Lane 1=Protein marker, Lane 3–6=Different fractions of bacterial protein expressed, Lane 7–10=His-tag purified recombinant cellulase like Chimeric protein.
See this image and copyright information in PMC

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