Characterization Amylase Gene Expression in Recombinant Escherichia coli Strain BL 21 and Rosetta

Nur Alisa, Kamarudin (2018) Characterization Amylase Gene Expression in Recombinant Escherichia coli Strain BL 21 and Rosetta. [Final Year Project Report] (Unpublished)

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Abstract

Amylase is endoenzyme, known as amyglucosidase, have importances in industrial especially in starchysaccharification - converting starch to glucose. The purpose of study were to optimize the expression ofheterologous gene in eukaryotes and characterize amylase production in recombinant Escherichia coli BL21 and Rosetta. The plasmid ofrecombinant Escherichia coli pFTag BL21 and Rosetta detected at 610 bp using DNA ladder Vivantis 1 kb. The qualitative analysis of gene amylase in pFTag BL21 and Rosetta be confirmed by the formation halo in iodine-starch hydrolysis method. Quantitive analysis of amylase production in this study is by using the method of Bradford for total protein estimation and DNS method for total of amylase in sample. The highest production of amylase was observed in Escherichia coli pFTag Bl2 l in the fermentation media after 5 days of incubation at room temperature and at pH 6.9 with dilution factor 0.1 X. Both host were treated with inducer IPTG and the production of amylase by host being compared with the sample without inducer. The highest record of total amylase in the sample with 100 ug/ml of inducer are from pFTAG BL21 with increasing 19.42 % from the wild type BL21 strain.

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