Proposed Implementation

● Material: Indigoidine

● Fermentation Method

   

Transfer the seed liquid to a 250mL conical flask containing 50mL fermentation medium at 1% inoculum size(V/V), in the early stage of the fermentation process, incubate at 37°C for 4-10h, incubate at 28°C for 12h in the late stage, the speed of the shaker during the whole fermentation process is 180r/min/.

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● Optimization of fermentation conditions

   

Optimization of medium nitrogen source:

   

According to the fermentation method of 2, the yield obtained is 0.35g/L. When tryptone:yeast extract=5:10 and 0:15, the highest output is 0.55g/L and 0.57g/L respectively. When tryptone:yeast extract=5:15, the yield is 0.44 g/L.

   

Optimization of Fermentation temperature change point:

   

The same proportion of peptone and yeast extract in the fermentation medium is significantly different at different temperature change points. When the temperature change point is 4h, peptone:yeast extract = 0:15, the maximum yield is 0.56g/L. At 6-8h,peptone:yeast extract=5:10, at 7h, the maximum yield is 1.4g/L.

   

● Conclusion

   

Fermentation medium composition：Tryptone 5g/L, yeast extract powder 10g/L. Final fermentation conditions: initial pH 7.0, culturing at 37°C for 7h, changing the temperature to 28°C for 12h.

● References

[1] 孙菁兰，王华敏，才 恒，王丽娟，唐巧巧，刘之毅，高 强 重组大肠杆菌产 Indigoidine 的发酵优化。

[2] Mohammad Rifqi Ghiffary, Cindy Pricilia Surya Prabowo, Komal Sharma, Yuchun Yan, Sang Yup Lee,* and Hyun Uk Kim* High-Level Production of the Natural Blue Pigment Indigoidine from Metabolically Engineered Corynebacterium glutamicum for Sustainable Fabric Dyes.

[3] Maren Wehrs, Jan‑Philip Prahl, Jadie Moon, Yuchen Li, Deepti Tanjore, Jay D. Keasling, Todd Pray and Aindrila Mukhopadhyay* Production efciency of the bacterial non-ribosomal peptide indigoidine relies on the respiratory metabolic state in S. cerevisiae.