YidC as a potential antibiotic target.

The membrane insertase YidC, is an essential bacterial component and functions in the folding and insertion of many membrane proteins during their biogenesis. It is a multispanning protein in the inner (cytoplasmic) membrane of Escherichia coli that binds its substrates in the "greasy slide" through hydrophobic interaction. The hydrophilic part of the substrate transiently localizes in the groove of YidC before it is translocated into the periplasm. The groove, which is flanked by the greasy slide, is within the center of the membrane, and provides a promising target for inhibitors that would block the insertase function of YidC. In addition, since the greasy slide is available for the binding of various substrates, it could also provide a binding site for inhibitory molecules. In this review we discuss in detail the structure and the mechanism of how YidC interacts not only with its substrates, but also with its partner proteins, the SecYEG translocase and the SRP signal recognition particle. Insight into the substrate binding to the YidC catalytic groove is presented. We wind up the review with the idea that the hydrophilic groove would be a potential site for drug binding and the feasibility of YidC-targeted drug development.

Bacterial Signal Peptides- Navigating the Journey of Proteins.

In 1971, Blobel proposed the first statement of the Signal Hypothesis which suggested that proteins have amino-terminal sequences that dictate their export and localization in the cell. A cytosolic binding factor was predicted, and later the protein conducting channel was discovered that was proposed in 1975 to align with the large ribosomal tunnel. The 1975 Signal Hypothesis also predicted that proteins targeted to different intracellular membranes would possess distinct signals and integral membrane proteins contained uncleaved signal sequences which initiate translocation of the polypeptide chain. This review summarizes the central role that the signal peptides play as address codes for proteins, their decisive role as targeting factors for delivery to the membrane and their function to activate the translocation machinery for export and membrane protein insertion. After shedding light on the navigation of proteins, the importance of removal of signal peptide and their degradation are addressed. Furthermore, the emerging work on signal peptidases as novel targets for antibiotic development is described.

Polarity/charge as a determinant of translocase requirements for membrane protein insertion.

The YidC insertase of Escherichia coli inserts membrane proteins with small periplasmic loops (~20 residues). However, it has difficulty transporting loops that contain positively charged residues compared to negatively charged residues and, as a result, increasing the positive charge has an increased requirement for the Sec machinery as compared to negatively charged loops (Zhu et al., 2013; Soman et al., 2014). This suggested that the polarity and charge of the periplasmic regions of membrane proteins determine the YidC and Sec translocase requirements for insertion. Here we tested this polarity/charge hypothesis by showing that insertion of our model substrate protein procoat-Lep can become YidC/Sec dependent when the periplasmic loop was converted to highly polar even in the absence of any charged residues. Moreover, adding a number of hydrophobic amino acids to a highly polar loop can decrease the Sec-dependence of the otherwise strictly Sec-dependent membrane proteins. We also demonstrate that the length of the procoat-Lep loop is indeed a determinant for Sec-dependence by inserting alanine residues that do not markedly change the overall hydrophilicity of the periplasmic loop. Taken together, the results support the polarity/charge hypothesis as a determinant for the translocase requirement for procoat insertion.

Silk Fibroin: An Emerging Biocompatible Material for Application of Enzymes and Whole Cells in Bioelectronics and Bioanalytical Sciences.

Enzymes and whole cells serve as the active biological entities in a myriad of applications including bioprocesses, bioanalytics, and bioelectronics. Conserving the natural activity of these functional biological entities during their prolonged use is one of the major goals for validating their practical applications. Silk fibroin (SF) has emerged as a biocompatible material to interface with enzymes as well as whole cells. These biomaterials can be tailored both physically and chemically to create excellent scaffolds of different forms such as fibers, films, and powder for immobilization and stabilization of enzymes. The secondary structures of the SF-protein can be attuned to generate hydrophobic/hydrophilic pockets suitable to create the biocompatible microenvironments. The fibrous nature of the SF protein with a dominant hydrophobic property may also serve as an excellent support for promoting cellular adhesion and growth. This review compiles and discusses the recent literature on the application of SF as a biocompatible material at the interface of enzymes and cells in various fields, including the emerging area of bioelectronics and bioanalytical sciences.

Composite polymer coated magnetic nanoparticles based anode enhances dye degradation and power production in microbial fuel cells.

Combined power generation and waste degradation through microbial fuel cell (MFC) technology is emerging as an attractive solution for controlling pollution in water bodies. Cyanobacteria as fuel cell catalysts for such shared energy activities are not well studied even though these possess robust metabolic systems supporting exo-electrogenicity, biodegradation of toxic compounds, and their survival under wide environmental conditions. Herein, a dual chambered (50 ml each) MFC assembled with Synechococcus sp. based bioanode and abiotic cathode for simultaneous power generation and Mordant orange dye degradation is reported. The anode was prepared by encrusting chemically synthesised magnetic nanoparticle (MNP) of size 8.4 ±â€¯0.2 nm with magnetization of 69 emu g-1on Toray carbon paper (TCP). The MNPs were encapsulated with aniline and pyrrole composite polymers to facilitate biofilm formation and cellular electron flow to the anode as confirmed by advance microscopic and voltametric techniques, respectively. The MFC with the dye mixed acetate produced current of 14.04 ±â€¯5.5 A m-3 with a maximum power density of 4.9 ±â€¯0.5 W m-3 (at cell voltage of 0.494 ±â€¯0.05 V), which was 18% higher than the control (without dye). The MFC produced a high OCP of 0.949 ±â€¯0.07 V and offered to decolorize 68.5% and degrade 89% of the dye following 216 h of its operation as confirmed by photometry (λ385 nm) and LC-MS/MS analyses, respectively. The efficient dye degradation is attributed to the bioanode for secreting high level of reactive oxygen species. The composite polymer coated MNPs anode with cyanobacterial biofilm is therefore, a highly efficient construct for enhanced azo dye degradation and associated power generation in a MFC system.

Bacterial Membrane Depolarization-Linked Fuel Cell Potential Burst as Signal for Selective Detection of Alcohol.

The biosensing application of microbial fuel cell (MFC) is hampered by its long response time, poor selectivity, and technical difficulty in developing portable devices. Herein, a novel signal form for rapid detection of ethanol was generated in a photosynthetic MFC (PMFC). First, a dual chambered (100 mL each) PMFC was fabricated by using cyanobacteria-based anode and abiotic cathode, and its performance was examined for detection of alcohols. A graphene-based nanobiocomposite matrix was layered over graphite anode to support cyanobacterial biofilm growth and to facilitate electron transfer. Injection of alcohols into the anodic chamber caused a transient potential burst of the PMFC within 60 s (load 1000 Ω), and the magnitude of potential could be correlated to the ethanol concentrations in the range 0.001-20% with a limit of detection (LOD) of 0.13% ( R2 = 0.96). The device exhibited higher selectivity toward ethanol than methanol as discerned from the corresponding cell-alcohol interaction constant ( Ki) of 780 and 1250 mM. The concept was then translated to a paper-based PMFC (p-PMFC) (size ∼20 cm2) wherein, the cells were merely immobilized over the anode. The device with a shelf life of ∼3 months detected ethanol within 10 s with a dynamic range of 0.005-10% and LOD of 0.02% ( R2 = 0.99). The fast response time was attributed to the higher wettability of ethanol on the immobilized cell surface as validated by the contact angle data. Alcohols degraded the cell membrane on the order of ethanol > methanol, enhanced the redox current of the membrane-bound electron carrier proteins, and pushed the anodic band gap toward more negative value. The consequence was the potential burst, the magnitude of which was correlated to the ethanol concentrations. This novel approach has a great application potential for selective, sensitive, rapid, and portable detection of ethanol.

Thin films of silk fibroin and its blend with chitosan strongly promote biofilm growth of Synechococcus sp. BDU 140432.

The activating role of different polymer thin films coated over polystyrene support on the Synechococcus sp. biofilm growth was examined concurrently by measuring biofilm florescence using a dye and by measuring cell density in the isolated biofilm. Compared to blank (no coating), the increase in biofilm formation (%) on silk, chitosan, silk-chitosan (3:2) blend, polyaniline, osmium, and Nafion films were 27.73 (31.16), 21.55 (23.74), 37.21 (38.34), 5.35 (8.96), 6.70 (6.55) and (nil), respectively with corresponding cell density (%) shown in the parentheses. This trend of biofilm formation on the films did not significantly vary for Escherichia coli and Lactobacillus plantarum strains. The films of 20 residues long each of glycine-alanine repeat peptide, which mimics a silk fibroin motif, and a hydrophobic glycine-valine repeat peptide, increased the biofilm growth by 13.53 % and 26.08 %, respectively. Silk and blend films showed highest adhesion unit (0.48-0.49), adhesion rate ((4.2-4.8)×10(-6), m/s) and Gibbs energy of adhesion (-8.5 to -8.6kT) with Synechococcus sp. The results confirmed interplay of electrostatic and hydrophobic interaction between cell-surface and polymer films for promoting rapid biofilm growth. This study established that the thin films of silk and the blend (3:2) promote rapid biofilm growth for all the tested microorganisms.