Tailoring the whole-cell sensing spectrum with cyborgian redox machinery.

Whole-cell biosensors are an important class of analytical tools that offer the advantages of low cost, facile operation, and unique reproduction/regeneration ability. However, it has always been quite challenging to expand the sensing spectrum of the host. Here, a new approach to extend the cell sensing spectrum with biomineralized nanoparticles is developed. The nano-biohybrid design comprise biomineralized FeS nanoparticles firmly anchored onto the bacterium, Shewanella oneidensis MR-1, wherein the nanoparticles are wired to the cellular electron transfer machinery (MtrCAB/OmcA) of the bacterium, forming an artificial cyborgian redox machinery consisting of FeS-MtrCAB/OmcA-FccA. Strikingly, with this cyborgian redox machinery, the sensing spectrum of FeS hybridized S. oneidensis MR-1 cell is successfully expanded to enable whole-cell electrochemical detection of Vitamin B12, while an unhybridized native cell is incapable of sensing. This proof-of-concept nano-biohybrid design offers a new perspective on manipulating the microbial toolkit for an expanded sensing spectrum in whole-cell biosensors.

Engineered cytochrome fused extracellular matrix enabled efficient extracellular electron transfer and improved performance of microbial fuel cell.

Microbial fuel cell (MFC) was a promising technology for energy harvesting from wastewater. However, inefficient bacterial extracellular electron transfer (EET) limited the performance as well as the applications of MFC. Here, a new strategy to reinforce the EET by engineering synthetic extracellular matrix (ECM) with cytochrome fused curli was developed. By genetically fusing a minimal cytochrome domain (MCD) with the curli protein CsgA and heterogeneously expressing in model exoelectrogen of Shewanella oneidensis MR-1, the cytochrome fused electroactive curli network was successfully constructed and assembled. Interestingly, the strain with the MCD fused synthetic ECM delivered about 2.4 times and 2.0 times higher voltage and power density output than these of wild type MR-1 in MFC. More impressively, electrochemical analysis suggested that this synthetic ECM not only introduced cytochrome of MCD, but also attracted more self-secreted electrochemically active substances, which might facilitate the EET and improve the MFC performance. This work demonstrated the possibility to manipulation the EET with ECM engineering, which opened up new path for exoelectrogen design and engineering.

Cytochrome C catalyzed oxygen tolerant atom-transfer radical polymerization.

Atom-transfer radical polymerization (ATRP) is a well-known technique for controlled polymer synthesis. However, the ATRP usually employed toxic heavy metal ionas as the catalyst and was susceptible to molecular oxygen, which made it should be conducted under strictly anoxic condition. Conducting ATRP under ambient and biocompatible conditions is the major challenge. In this study, cytochrome C was explored as an efficient biocatalyst for ATRP under biocompatible conditions. The cytochrome C catalyzed ATRP showed a relatively low polymer dispersity index of 1.19. More interestingly, the cytochrome C catalyzed ATRP showed superior oxygen resistance as it could be performed under aerobic conditions with high dissolved oxygen level. Further analysis suggested that the Fe(II) embed in the cytochrome C might serve as the catalytic center and methyl radical was responsible for the ATRP catalysis. This work explored new biocompatible catalyst for aerobic ATRP, which might open new dimension for practical ATRP and application of cytochrome C protein.