Experiments

Experiment

Design

Team Seoul_Korea assured that the cellulose amount in decomposed pine tree bark decreases and that it can successfully and effectively absorb harmful oil. However, waiting for dissolution by microbes takes way too long and the outcome is far less than the amount needed to absorb oil. Also, as human labor is needed to find dissolved oil, the method takes up too much resources and time. Therefore, Seoul_Korea aimed to produce a sorbent using pine trees with synthetic biology. First, by examining documents, Seoul_Korea found 4 kinds of cellulose degrading enzymes (Cellobiohydrolase I, II, beta-glucosidase, endoglucanase) that have originated from bacteria or heterologous expression checked. (Figure 2). Also, to simplify the process of cellulose dissolving, Seoul_Korea designed the experiment for signal peptides to fusion so that the enzymes could secrete. Seoul_Korea randomly inserted 12 kinds of secretion signals and effectively found out the secretion signals of secretied clones by dot blot method to find the most suitable signal peptide of each enzyme. Seoul_Korea applied the IPTG inducible system to express mass amounts of enzymes in E. coli and Bacillus subtilis. Through this process, enzymes secreted by media effectively dissolved cellulose, and Seoul_Korea confirmed that using this method, it takes 1/10 of the time to make a sorbent from our hardware system compared to nature producing the same product.

Protocols