Team:TPR China/Composite

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pTac_ AdNIT(BBa_K3599009)

We decided to use nitrilase - a protein that degrades nitriles - for degradation of PAN.
According to the literature,we selected nitrilase with high enzymatic activity and relatively complete sequences - AdNIT(BBa_K3599003) and successfully constructed pTac_AdNIT(BBa_K3599009).

Design of pTac_AdNIT

Our effector system is construct on Pet28-b plasmid and bind with upstream LacI sequence. . We transformed pTac_AdNIT vector into E .coli BL21 for expression.

Nitrilase Enzyme Activity

The activity of nitrilase was verified by detecting the ammonium ions produced by the degradation of PAN by the bacterial liquid. We define U as the amount of PAN that can be degraded per microliter per hour.
Fig.1 Experimental phenomena of enzyme activity test
Fig.2 Enzymatic activity of Nitrilase
As can be seen from Fig.2, the U value of pTac_LaNIT is 0.6115, which is 1.27 times that of the negative control;the U value of pTac_AdNIT is 0.6205, which is 1.3 times that of the negative control.These data indicate that our engineered bacteria have high activity of nitrilase.