17 A. Muazzam 2 B. Malik N. Rashid S. Irshad M. Fatima Journal of Animal and Plant Sciences JAPS 20192019/02/01 29 1 99-108 1018-7081 N/A <p>Alpha amylase (&alpha;-amylase) from<em>&nbsp;Bacillus licheniformis</em>&nbsp;shows great role in starch hydrolysis for industrial application due to its thermostability, long half-life and specificity. However, it is difficult to produce this enzyme in large quantities because of formation of inclusion bodies, but lowering the temperature to 18 &deg;C leads to a soluble expression of protein. This also&nbsp;highlighted the efficacy of&nbsp;<em>Escherichia coli</em>&nbsp;as one of the best host for secretion of recombinant proteins. A further study to find an approach for a soluble expression of &alpha;-amylase was conducted here, using pET-22b(+)&nbsp; as an expression vector and temperature optimization. Purified monomeric enzyme displayed a specific activity of 584.61 U/mg at 37 &deg;C and a molecular mass of 52&nbsp;<em>kDa</em>. Purification fold of recombinant &alpha;-amylase was 2.04 times in comparison to crude amylase extract.&nbsp; Relative activity of enzymewas remarkably enhanced by Ca+2 but strongly impeded by Cu2+ and Na+1, highlighted the significance of Ca+2 as a cofactor for optimal amylase activity.</p> Hydrolysis, Soluble, Thermostable, Industrial Pakistan Agricultural Scientists Forum https://thejaps.org.pk/AbstractView.aspx?mid=Bioch-17-0026