Team:ASIJ Tokyo/Improve

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Improve

Building on previous projects and parts

To provide alpha amylase for creating models to simulate salivary α-amylase conditions, we created a new construct designed to actively produce α-amylase. This would enable others to produce α-amylase for the purposes of testing α-amylase inhibition.

Thus, our team’s new construct BBa_K2928015 has been designed to produce similar concentrations of α-amylase to human saliva. Based on the existing part BBa_K523006, we experimented with the promoter strengths and were able to produce a similar concentration of amylase in vitro. As clearly visible in the graph below, the Anderson J23101 promoter (BBa_J23101) produced a concentration of α-amylase within the margin of that in human saliva. Moreover, using the Anderson J23101 promoter, the e coli produced 1.002900 mg/ml of α-amylase, compared to the range of 0.84–4.44 mg/ml of α-amylase in human saliva. Using this construct, we believe future teams can simulate α-amylase conditions in human saliva.
Figure 1. After testing several promoters, we concluded the Anderson J23101 promoter produced 1.0029 mg/ml of amylase, closest to the 2.64 mg/ml (±1.8) in average human saliva.