Molecular-level understanding of the adsorption mechanism of a graphite-binding peptide at the water/graphite interface.

The association of proteins and peptides with inorganic material has vast technological potential. An understanding of the adsorption of peptides at liquid/solid interfaces on a molecular-level is fundamental to fully realising this potential. Combining our prior work along with the statistical analysis of 100+ molecular dynamics simulations of adsorption of an experimentally identified graphite binding peptide, GrBP5, at the water/graphite interface has been used here to propose a model for the adsorption of a peptide at a liquid/solid interface. This bottom-up model splits the adsorption process into three reversible phases: biased diffusion, anchoring and lockdown. Statistical analysis highlighted the distinct roles played by regions of the peptide studied here throughout the adsorption process: the hydrophobic domain plays a significant role in the biased diffusion and anchoring phases suggesting that the initial impetus for association between the peptide and the interface may be hydrophobic in origin; aromatic residues dominate the interaction between the peptide and the surface in the adsorbed state and the polar region in the middle of the peptide affords a high conformational flexibility allowing strongly interacting residues to maximise favourable interactions with the surface. Reversible adsorption was observed here, unlike in our prior work focused on a more strongly interacting surface. However, this reversibility is unlikely to be seen once the peptide-surface interaction exceeds 10 kcal mol(-1).

Biological response on a titanium implant-grade surface functionalized with modular peptides.

Titanium (Ti) and its alloys are among the most successful implantable materials for dental and orthopedic applications. The combination of excellent mechanical and corrosion resistance properties makes them highly desirable as endosseous implants that can withstand a demanding biomechanical environment. Yet, the success of the implant depends on its osteointegration, which is modulated by the biological reactions occurring at the interface of the implant. A recent development for improving biological responses on the Ti-implant surface has been the realization that bifunctional peptides can impart material binding specificity not only because of their molecular recognition of the inorganic material surface, but also through their self-assembly and ease of biological conjugation properties. To assess peptide-based functionalization on bioactivity, the present authors generated a set of peptides for implant-grade Ti, using cell surface display methods. Out of 60 unique peptides selected by this method, two of the strongest titanium binding peptides, TiBP1 and TiBP2, were further characterized for molecular structure and adsorption properties. These two peptides demonstrated unique, but similar molecular conformations different from that of a weak binder peptide, TiBP60. Adsorption measurements on a Ti surface revealed that their disassociation constants were 15-fold less than TiBP60. Their flexible and modular use in biological surface functionalization were demonstrated by conjugating them with an integrin recognizing peptide motif, RGDS. The functionalization of the Ti surface by the selected peptides significantly enhanced the bioactivity of osteoblast and fibroblast cells on implant-grade materials.

Multifunctional protein-enabled patterning on arrayed ferroelectric materials.

This study demonstrates a biological route to programming well-defined protein-inorganic interfaces with an arrayed geometry via modular peptide tag technology. To illustrate this concept, we designed a model multifunctional fusion protein, which simultaneously displays a maltose-binding protein (MBP), a green fluorescence protein (GFPuv) and an inorganic-binding peptide (AgBP2C). The fused combinatorially selected AgBP2C tag controls and site-directs the multifunctional fusion protein to immobilize on silver nanoparticle arrays that are fabricated on specific domain surfaces of ferroelectric LiNbO(3) via photochemical deposition and in situ synthesis. Our combined peptide-assisted biological and ferroelectric lithography approach offers modular design and versatility in tailoring surface reactivity for fabrication of nanoscale devices in environmentally benign conditions.

Methanogenic and sulphate reducing bacterial population levels in a full-scale anaerobic reactor treating pulp and paper industry wastewater using fluorescence in situ hybridisation.

In this study, specific methanogenic activity (SMA) test and fluorescence in situ hybridisation (FISH) were respectively used to determine acetoclastic methanogenic capacity, and composition and number of methanogenic and sulphate reducing bacterial (SRB) populations within a full scale anaerobic contact reactor treating a pulp and paper industry effluent. The sludge samples were collected from three different heights along the anaerobic reactor having a difficulty of completely stirring. Performance of the anaerobic reactor in terms of COD removal efficiency varied between 47 and 55% at organic loading rates in a range of 1.6-1.8 kg COD m(-3) d(-1) and methane yield varied between 0.18 and 0.20 m3CH4kg CODrem(-1). The anaerobic reactor was not operated for 2 weeks during the monitoring period. According to SMA test results, potential methane production rate was 276 mLCH4 gVSS(-1) d(-1) before the off period of the reactor, however it decreased to 159 mL CH4 gVSS(-1) d(-1) after this period. SMA test and FISH results along the reactor height showed that the acetoclastic methanogenic activity of the sludge samples, the relative abundance of acetoclastic methanogens, hydrogenotrophic methanogens and acetate oxidising SRB decreased as the reactor height increased, however the relative abundance of non-acetate oxidising SRB increased.

Respirometric evaluation and modeling of glucose utilization by Escherichia coli under aerobic and mesophilic cultivation conditions.

The study presents a mechanistic model for the evaluation of glucose utilization by Escherichia coli under aerobic and mesophilic growth conditions. In the first step, the experimental data was derived from batch respirometric experiments conducted at 37 degrees C, using two different initial substrate to microorganism (S(0)/X(0)) ratios of 15.0 and 1.3 mgCOD/mgSS. Acetate generation, glycogen formation and oxygen uptake rate profile were monitored together with glucose uptake and biomass increase throughout the experiments. The oxygen uptake rate (OUR) exhibited a typical profile accounting for growth on glucose, acetate and glycogen. No acetate formation (overflow) was detected at low initial S(0)/X(0) ratio. In the second step, the effect of culture history developed under long-term growth limiting conditions on the kinetics of glucose utilization by the same culture was evaluated in a sequencing batch reactor (SBR). The system was operated at cyclic steady state with a constant mean cell residence time of 5 days. The kinetic response of E.coli culture was followed by similar measurements within a complete cycle. Model calibration for the SBR system showed that E. coli culture regulated its growth metabolism by decreasing the maximum growth rate (lower microH) together with an increase of substrate affinity (lower K(S)) as compared to uncontrolled growth conditions. The continuous low rate operation of SBR system induced a significant biochemical substrate storage capability as glycogen in parallel to growth, which persisted throughout the operation. The acetate overflow was observed again as an important mechanism to be accounted for in the evaluation of process kinetics.

Effect of pH on physiology of Metarhizium anisopliae for production of swainsonine.

The effect of pH on the production of swainsonine and fungal morphology at different stages of fermentation of Metarhizium anisopliae was investigated. When no control was applied, the pH of the culture dropped from 6.5 to 3.8 within the first 72 hours and the concentration of swainsonine reached 43.3 mg 1(-1). When the pH was held constant either at the beginning or throughout the fermentation, the maximum recorded swainsonine level was only 8.4 mg 1(-1) corresponding with an increase in the formation of pellets. A late pH control applied after 72 hours, resulted in a swainsonine titer of 45.5 mg 1(-1).

Enhanced penicillin production by oligosaccharides from batch cultures of Penicillium chrysogenum in stirred-tank reactors.

Alginate and galactomannan-derived oligosaccharides enhanced the production of penicillin G when added to stirred tank reactor cultures of Penicillium chrysogenum. The addition of oligomannuronate and oligoguluronate blocks increased penicillin G yield by 47% and 49%, respectively. The effect of mannan oligosaccharides was found to be more pronounced with 69% higher yield than the control cultures. The maximum increase in the average specific productivity of the oligosaccharide augmented cultures was 55% after addition of mannan oligosaccharides. In addition, a difference was observed in all cases in the accumulation pattern of the intermediate of penicillin biosynthesis, delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine.