Team:JNFLS/Proof Of Concept
All the current application research of bacterial cellulose is based on the organisms that originally synthesize bacterial cellulose, such as acetobacteria xylinus. Most researchers are only optimizing the fermentation conditions in order to obtain a larger yield of bacterial cellulose. Considering that bacterium such as acetobacteria xylinus with the ability to synthesize cellulose is not an engineered strain, its fermentation conditions are difficult to control. And the preservation, recovery, and activation process of acetobacteria xylinus are tedious, and the ability of cellulose production is easy to fall. Our project is to try to clone the genes related to the synthesis of bacterial cellulose from acetobacteria xylinus, and transfer them to the engineered E. coli, which fermentation conditions are experienced, preservation is convenient, and the industrial production is facilitated to be realized.
1.We have cloned 7 genes successfully from Acetobacter xylinus, and transferred them into the E. coli
We have cloned seven related genes, four of which are directly related to the synthesis of bacterial cellulose; one is a protein that helps cellulose synthase locate on the membrane; two of them are also enzymes that help straighten out the sugar chain.
To avoid repetition, please refer to our results:https://2020.igem.org/Team:JNFLS/Results
2.The E. coli transferred with genes related to the synthesis of bacterial cellulose works already
After completion gene cloning work, the transferred E. coli were cultured in the medium for bacterial cellulose production. We used the optimized conditions acquired from the acetobacter xylinum culturing for producing bacterial cellulose. The E. coli were cultured for 7days at 30°C, pH6.8, 10% inoculum size, and glucose as the carbon source, then the cellulose in the medium was harvested. The result is showed in the following figure.
The production of bacterial cellulose in transferred E. coli.
Summary:
The result showed that the bacterial cellulose was produced successfully in the transferred E. coli, indicating that the parts transferred to the E. coli worked well. The result also showed that the production yield is not as high as in the acetobacter xyilus (showed in the result of our project on the wiki), suggesting that the culture conditions still need optimization.
For more information, please refer to our engineering:https://2020.igem.org/Team:JNFLS/Engineering