The rapid expansion of the fast fashion market has led to overproduction and overconsumption of cheap, disposable clothes. However, the majority of them end up as textile waste at incineration plants or landfill sites, exacerbating the global climate problem. To minimize the environmental pollution of the fashion industry, GreatBay_SCIE proposed a sustainable and eco-friendly method to upcycle unwanted clothes using the cellulosome-like complex nanomachines. These protein complexes are composed of scaffold proteins OlpB and CipA, with cellulolytic enzymes and PET-degrading enzymes fused on them to enable the effective biodegradation of cellulose fiber and PET fiber in clothes, converting them into reusable end products including bacterial cellulose (BC) and PET beads. With Fabrevivo, we aim to revolutionize textile recycling, making fast fashion more environmentally-friendly.
Fig. 1 The schematic representation of Fabrevivo design. Two cellulosome complexes were constructed, with cellulase and PETase subunits binding to primary scaffold CipA1B2C via type I cohesin-dockerin interaction; Attachment of CipA scaffold to OlpB-Ag3 was made possible by type II cohesin-dockerin interaction, Neae-Nb3-Ag3 association displays the whole complex on _E. coli_ surface. Ferritin expression within _E. coli_ host enables magnetic recycling for cellulosome complexes to be reused.
#### Surface display system
## Surface display system
In order to display the cellulosome complex on the surface of _E. coli_, we decided to use nanobody (Nb)-antigen (Ag) interaction between Neae-Nb3 and Ag3.
@@ -36,7 +36,7 @@ The fungal cellulases we used were produced as fusion proteins ligated with type
#### **Assembling cellulosome-like complexes displayed on the surface of _E. coli_ and their superior ability to degrade cellulose.**
We assembled the cellulose-like complex (Celly complex) on the surface of _E.coli_ and added cellulose. After 24h, the mixture was filtered and tested for glucose by Benedict's test. From the result, we determined that the cellulosome-like complexes were able to degrade cellulose at a higher efficiency than the cell-free cellulases mixture(Fig.4A), with the cellulosome group producing 27.85 more glucose than the cell-free cellulases+cellulase boosters group, suggested by the value of the sample absorbance at 635 nm(Fig.4B).
We assembled the cellulose-like complex (Celly complex) on the surface of E.coli and added cellulose. After 24h, the mixture was filtered and tested for glucose by Benedict's test. From the result, we determined that the cellulosome-like complexes were able to degrade cellulose at a higher efficiency than the cell-free cellulases mixture(Fig.4A), which is proven by the apparent red color of Benedict's test that indicates higher concentration of reducing sugar in cellulosome sample. This conclusion was also supported by the measure of light absorbance at 635nm in each sample (Fig.4B).
