MODEL

Purpose of the experiment

In order to study the growth pattern of bacteria and the growth rate of Biotinylated Staphylococcus aureus (S. aureus), we divided S. aureus into two groups, one with None treatment, where 1 ml of extracted lysate was taken from the cells at OD 0.3 of the bacteria. The other group was the bacteria after blocking with TurboID-AIP biotin, and the lysate was still extracted at OD 0.3. The growth of S. aureus in leftovers and the effect of TurboID-AIP biotin on the growth rate of S. aureus were quantified by calculating the S. aureus growth rate index.

Materials:

1. TurboID-N-AgrD-AIP protein extraction from BS_United_China.
2. 50 mM Biotin
3. 100 mM ATP
4. S. aureus in Hainan university.
5. Biotinylated S. aureus: generated by incubation with 200 ul TurboID-N-AgrD-AIP protein extraction, 500 uM Biotin, 1 mM ATP for 1 hours at room temperature.

BS_United_China team designed the experiments. Protocols are designed by BS_United_China. All the explements related to culture S. aureus had done by the staff members from Prof. Yandu Lu in Hainan university. Video were recorded in Prof. Yandu Lu’s lab. BS_United_China team collected the data remotely and did the followed analysis and calculations.

Procedure

1. Use OD600 to measure the growth rate of S. aureus with different biotin synthesis
2. Calculate the average growth rate of each measure unit
3. Use the data to make models of the average growth rate in OD/hour

Part 1

Fig.1 The comparison of the S. aureus with TurboID (A)&(C) and S. aureus with TurboID-AgrD-AIP+Dethiobiotin synthetase & Biotin synthase (B)&(D)

In this group of graphs, we compare the average growth rate of each data, and can apparently concluded that the average growth rate of the S. aureus with TurboID-AgrD-AIP+Dethiobiotin synthetase & Diotin synthase is much smaller of the the average growth rate of the S. aureus with TurboID only.

Part 2

Fig.2 The comparation of the S. aureus with TurboID (A)&(C) and the None treatment (B)&(D)

In this group of graphs, we compare the average growth rate of each data, and can apparently concluded that the average growth rate of the S. aureus with TurboID is much smaller than the S. aureus with none treatment.

Conclusion of part1&2

The comparations in the part1 and part2 lead directly to the conclusion that the S. aureus with our biotin is effective, between our treatments the S. aureus with TurboID-AgrD-AIP+Dethiobiotin synthetase & Diotin synthase is more effective than the S. aureus with TurboID only.

Part 3

To further validate the inhibitory effect of TurboID-AIP and Biotin on environmental Staphylococcus aureus, we used a gradient test.

Fig.3 Analysis of the data fluorescence value of the soil extract drops (A) and made a model (B)

Conclusion of part3

This linear relationship indicates that after titrating 25ul, the inhibitory effect of this biotin on S. aureus became obvious and the effect increased geometrically. At the same time, because the bacterial content of the soil is very high, if the soil sample is changed to fresh seafood, then the amount of drops added may be smaller, and it does not even take 25ul to achieve a very significant inhibitory effect.

Conclusion

Our biotins can successfully inhibit the growth of the S. aureus even in a small quantity.