Team:AHUT China/Demonstrate

Demonstrate

Our project can be applied in our designed simulation model for CO2 capture, which might be suitable for industrial production.

Last year, we successfully constructed a biobrick CA2 (L203K) (BBa_K2547004) to optimize the thermal stability of wild-type carbonic anhydrase, but the low reuse ability of CA2 (L203K) limited further industrial applications.

This year, we further optimized CA2 (L203K), and obtained the immobilized carbonic anhydrase for efficient CO2 capture.

We have constructed and tested the CO2 capture system based on pET-30a (+) expression vector and E. coli TB1 strain under simulated condition in laboratory.

The immobilized CA2(L203K)-C-LCTPSR protein have higher activity than immobilized CA2(L203K)-N-LCTPSR protein, so we designed a simulate model to absorb CO2 in industrial waste gas for measuring the reuse ability of CA2(L203K)-C-LCTPSR.

Our main achievements

1. We constructed new biobricks [CA2(L203K)-C-LCTPSR and CA2(L203K)-N-LCTPSR].

2. The CA2(L203K)-C-LCTPSR and CA2(L203K)-N- LCTPSR protein were immobilized.

3. The immobilized CA2(L203K)-C-LCTPSR protein have higher activity and reuse ability than immobilized CA2(L203K)-N-LCTPSR protein.

4. The reuse ability of the immobilized CA2(L203K)-C-LCTPSR protein was tested by our designed simulation model.

5.Immobilized CA2(L203K)-C-LCTPSR protein showed potential reuse ability.

Measurment of Enzyme activity

To demonstrate that the CA2(L203K)-C-LCTPSR and CA2(L203K)-N-LCTPSR protein retain the activity of the high enzymatic activity, CA2(L203K)-C-LCTPSR and CA2(L203K)-N-LCTPSR protein were tested experimentally by esterase activity assay at 37℃ and 50℃.

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Fig.1 Esterase activity analysis of free CA2(L203K), free CA2(L203K)-C-LCTPSR and immobilized CA2(L203K)-C-LCTPSR protein at 37℃
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Fig.2 Esterase activity analysis of free CA2(L203K), free CA2(L203K)-C-LCTPSR and immobilized CA2(L203K)-C-LCTPSR protein at 50℃
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Fig.3 Esterase activity analysis of immobilized CA2(L203K)-N-LCTPSR protein at 37℃ and 50℃

As shown in Fig.1 and Fig.2 as well as Fig.3, CA2 (L203K)-C-LCTPSR and CA2(L203K)-N-LCTPSR protein were stable at high temperature and retained their activity.

The reuse ability

The reuse ability of the immobilized enzyme was tested by our designed simulation model (Fig. 4).

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Fig.4 Simulation Model
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Fig.5 The reuse ability of CO2 capture of the immobilized CA2(L203K)-C-LCTPSR under our designed model.

The result showed that the immobilized CA2(L203K)-C-LCTPSR protein could absorb more CO2 under the simulation model and showed potential reuse ability as indicated in Fig.5.

CATALOGUE

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