Increase Production of the Enzyme by Cell Genetic Modification

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Faghihi, Farhad | 2020

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Type of Document: M.Sc. Thesis
Language: Farsi
Document No: 53181 (06)
University: Sharif University of Technology
Department: Chemical and Petroleum Engineering
Advisor(s): Roosta Azad, Reza; Banaei Moghaddam, Ali Mohammad
Abstract:
Genetic engineering of cells has been one of the most effective ways to increase protein production in wild strains. Among the various genetic engineering techniques, the CRISPR/Cas9 system that directly edits the cell chromosome, yielding greater efficiency and easier operation. In this study, we used this system to modify the genome of Bacillus subtilis ATCC 6633 to increase the production of extracellular alkaline protease enzyme. Alkaline protease enzyme has many applications in different industries, but in this study, the application of this enzyme in the detergent industry has been considered. To increase the production of this enzyme, we disrupted and prevented the gene responsible production of the ScoC protein (A regulatory protein that inhibits the transcription of the alkaline protease gene). The enzymatic evaluation showed that cell genome modification increased 2.82-fold production of alkaline protease in recombinant strain compared to the wild-type strain
Keywords:
Bacillus Subtilis ; Alkaline Protease ; Increase Protein Production ; Genetic Engineering ; Industrial Strain ; Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)System ; Genome Editing

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