Characterization and use of a bacterial lignin peroxidase with an improved manganese-oxidative activity.

Peroxidases are well-known biocatalysts produced by all organisms, especially microorganisms, and used in a number of biotechnological applications. The enzyme DypB from the lignin-degrading bacterium Rhodococcus jostii was recently shown to degrade solvent-obtained fractions of a Kraft lignin. In order to promote the practical use, the N246A variant of DypB, named Rh_DypB, was overexpressed in E. coli using a designed synthetic gene: by employing optimized conditions, the enzyme was fully produced as folded holoenzyme, thus avoiding the need for a further time-consuming and expensive reconstitution step. By a single chromatographic purification step, > 100 mg enzyme/L fermentation broth with a > 90% purity was produced. Rh_DypB shows a classical peroxidase activity which is significantly increased by adding Mn2+ ions: kinetic parameters for H2O2, Mn2+, ABTS, and 2,6-DMP were determined. The recombinant enzyme shows a good thermostability (melting temperature of 63-65 °C), is stable at pH 6-7, and maintains a large part of the starting activity following incubation for 24 h at 25-37 °C. Rh_DypB activity is not affected by 1 M NaCl, 10% DMSO, and 5% Tween-80, i.e., compounds used for dye decolorization or lignin-solubilization processes. The enzyme shows broad dye-decolorization activity, especially in the presence of Mn2+, oxidizes various aromatic monomers from lignin, and cleaves the guaiacylglycerol-ß-guaiacyl ether (GGE), i.e., the Cα-Cß bond of the dimeric lignin model molecule of ß-O-4 linkages. Under optimized conditions, 2 mM GGE was fully cleaved by recombinant Rh_DypB, generating guaiacol in only 10 min, at a rate of 12.5 µmol/min mg enzyme.

Kinetic characterization, thermo-stability and Reactive Red 195A dye detoxifying properties of manganese peroxidase-coupled gelatin hydrogel.

An indigenous and industrially important manganese peroxidase (MnP) was isolated from solid-state bio-processing of wheat bran by white-rot fungal strain Ganoderma lucidum IBL-05 under pre-optimized growth conditions. Crude MnP extract was partially purified (2.34-fold) to apparent homogeneity by ammonium sulphate precipitation and dialysis. The homogeneous enzyme preparation was encapsulated on gelatin matrix using glutaraldehyde as a cross-linking agent. Optimal conditions for highest immobilization (82.5%) were: gelatin 20% (w/v), glutaraldehyde 0.25% (v/v) and 2 h activation time using 0.6 mg/mL of protein concentration. Gelatin-encapsulated MnP presented its maximum activity at pH 6.0 and 60 °C. Thermo-stability was considerably improved after immobilization. The optimally active MnP fraction was tested against MnSO4 as a substrate to calculate kinetic parameters. More than 90% decolorization of Sandal-fix Red C4BLN (Reactive Red 195A) dye was achieved with immobilized MnP in 5 h. It also preserved more than 50% of its original activity after the sixth reusability cycle. The water quality parameters (pH, chemical oxygen demand, total organic carbon) and cytotoxicity (brine shrimp and Daphnia magna) studies revealed the non-toxic nature of the bio-treated dye sample. A lower Km, higher Vmax, greater acidic and thermal-resistant up to 60 °C were the improved catalytic features of immobilized MnP suggesting its suitability for a variety of biotechnological applications.

Characterization of Titratable Amphiphiles in Lipid Membranes by Fluorescence Spectroscopy.

Understanding the ionization behavior of lipid membranes is a key parameter for successful development of lipid-based drug delivery systems. Accurate determination of the ionization state of a titratable species incorporated in a lipid bilayer however requires special care. Herein we investigated the behavior of titratable lipids in liposomes by fluorescence spectroscopy and determined which extrinsic parameters-i.e., besides those directly related to their molecular structure-determine their ionization state. Two fluorescent dyes, TNS and R18, have been used to investigate basic and acidic titratable lipids, respectively. Our results suggest that the titration behavior of the ionizable lipid in the membrane is more sensitive to the composition of the membrane and to its physical state than to the presence of solutes in the aqueous phase. Essentially overlooked in earlier studies on ionizable lipid assemblies, the concentration of the titratable lipid in the membrane was found to have a major effect on the ionization state of the lipid polar head. This may result in a shift in the apparent pKa value which may be as large as two pKa units and cannot be satisfactorily predicted.

Excited-state proton transfer of weak photoacids adsorbed on biomaterials: proton transfer to glucosamine of chitosan.

UV-vis steady-state and time-resolved techniques were employed to study the excited-state proton-transfer process from two weak photoacids positioned next to the surface of chitosan and cellulose. Both chitosan and cellulose are linear polysaccharides; chitosan is composed mainly of d-glucosamine units. In order to overcome the problem of the high basicity of the glucosamine, we chose 2-naphthol (pKa* ≈ 2.7) and 2-naphthol-6-sulfonate (pKa* ≈ 1.7) as the proton emitters because of their ground state pKa (≈9). Next to the 1:1 cellulose:water weight ratio, the ESPT rate of these photoacids is comparable to that of bulk water. We found that the ESPT rate of 2-naphthol (2NP) and 2-naphthol-6-sulfonate (2N6S) next to chitosan in water (1:1) weight ratio samples is higher than in bulk water by a factor of about 5 and 2, respectively. We also found an efficient ESPT process that takes place from these photoacids in the methanol environment next to the chitosan scaffold, whereas ESPT is not observed in methanolic bulk solutions of these photoacids. We therefore conclude that ESPT occurs from these photoacids to the d-glucosamine units that make up chitosan.

Decolorization of dyeing wastewater and characterization of flocs during coagulation by a new composite coagulant.

A composite coagulant, polymeric aluminum ferric chloride-poly-epichlorohydrin-ethylenediamine (PAFC-EPI-ETA), was synthesized and then used for the treatment of synthetic reactive brilliant red (RBR) dyeing wastewater. Effects of (Fe+Al) to EPI-ETA mass ratio (P) on the color removal and zeta potential were investigated under different coagulant dosages and initial pHs. Experimental results indicate that the removal of reactive dye and the charge neutralization ability of composite coagulant were improved by increasing the content of organic EPI-ETA. PAFC-EPI-ETA with P=1 achieved the best color removal percentage and strongest charge neutralization ability. Decolorization efficiency using PAFC-EPI-ETA was quite effecient with pH range of 6.0-7.5 for RBR dye removal. Characteristics of the formed floc were investigated with the dosage of PAFC-EPI-ETA under different P values. The low (Fe+Al) to EPI-ETA mass ratio contributed to the formation of flocs with high growth rate, large size and large size variation.

[Colorimetric detection of coxsackievirus A6 by reverse transcription loop mediated isothermal amplification].

OBJECTIVE: To develop a simple, rapid and sensitive colorimetric reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of coxsackievirus A6 (CV-A6) based on the colour chang of hydroxy naphthol blue (HNB). METHODS: The method employed a set of six primers that recognized sequences of VP1 gene for amplification of nucleic acid under isothermal conditions at 63 °C for 50 min. The products were detected through visual inspection of color change by the pre-addition of HNB dye. The specificity was validated by detecting a collection of different human enteroviruses. The sensitivity of this assay was evaluated by serial dilutions of RNA molecules from in vitro transcription of CV-A6 VP1 gene, and compared with real-time RT-PCR (rRT-PCR) in parallel. This assay was evaluated with 92 clinical specimens from patients with hand-foot-mouth disease. RESULTS: A positive color (sky blue) was only observed in the preparation of CV-A6, whereas none of the other 23 kinds of human enteroviruses showed a color change. The HNB based RT-LAMP showed a sensitivity of 100 copies/reaction, which was at the same level as that of the rRT-PCR. The result of RT-LAMP in analysis of 92 clinical specimens was consistent with that of the rRT-PCR. The kappa correlation between the two methods was 1 and both of the sensitivity and specificity of the RT-LAMP assay were 100%. CONCLUSION: The established RT-LAMP assay had good specificity and sensitivity and thus demonstrated to be a promising screening tool for CV-A6 infection. It also has the potential to be used in resource-limited clinical sites and field study.

Potential use of low-cost lignocellulosic waste for the removal of direct violet 51 from aqueous solution: equilibrium and breakthrough studies.

An efficient biosorbent, sugarcane bagasse was used in native, HCl-treated, and Na-alginate immobilized form for the removal of Direct Violet 51 dye from aqueous solutions. Batch study was performed to optimize important process parameters, such as pH, contact time, biosorbent dose, initial dye concentration, and temperature. Removal of Direct Violet 51 was found to be favorable at pH 2 with the biosorbent dose of 0.05 g. Biosorption process was found to be exothermic in nature. Maximum dye biosorption (39.6 mg/g) was achieved by using HCl-treated biomass. The pseudo-second-order kinetic and Langmuir adsorption isotherm models showed best fitness to the experimental data. Thermodynamic study was also performed to determine the feasibility of biosorption process. Continuous mode study was performed to optimize the important process parameters, such as bed height, flow rate, and initial dye concentration for maximum removal of Direct Violet 51 dye. The higher bed height, low flow rate, and high initial dye concentration were found to be the better conditions for maximum dye biosorption (17.28 mg/g). The linearized form of the Thomas model equation fitted well to the experimental data. The bed depth service time model was used to express the effect of bed height on breakthrough curves. Characterization of biosorbent was performed by scanning electron microscopy and Fourier transform infrared (FT-IR) analysis. The FT-IR spectral analyses showed the involvement of hydroxyl, carbonyl, and carboxyl groups in biosorption process. These results indicated that sugarcane bagasse biomass could be used as a novel biosorbent for the removal of Direct Violet 51 dye from real textile and related industries.

Temperature compensation in determining of Remazol black B concentrations using plastic optical fiber based sensor.

In this study, the construction and test of tapered plastic optical fiber (POF) sensors, based on an intensity modulation approach are described. Tapered fiber sensors with different diameters of 0.65 mm, 0.45 mm, and 0.35 mm, were used to measure various concentrations of Remazol black B (RBB) dye aqueous solutions at room temperature. The concentrations of the RBB solutions were varied from 0 ppm to 70 ppm. In addition, the effect of varying the temperature of the RBB solution was also investigated. In this case, the output of the sensor was measured at four different temperatures of 27 °C, 30 °C, 35 °C, and 40 °C, while its concentration was fixed at 50 ppm and 100 ppm. The experimental results show that the tapered POF with d = 0.45 mm achieves the best performance with a reasonably good sensitivity of 61 × 10(-4) and a linearity of more than 99%. It also maintains a sufficient and stable signal when heat was applied to the solution with a linearity of more than 97%. Since the transmitted intensity is dependent on both the concentration and temperature of the analyte, multiple linear regression analysis was performed to combine the two independent variables into a single equation. The resulting equation was then validated experimentally and the best agreement between the calculated and experimental results was achieved by the sensor with d = 0.45 mm, where the minimum discrepancy is less than 5%. The authors conclude that POF-based sensors are suitable for RBB dye concentration sensing and, with refinement in fabrication, better results could be achieved. Their low fabrication cost, simple configuration, accuracy, and high sensitivity would attract many potential applications in chemical and biological sensing.

Fixed bed adsorption of 2-naphthalenesulfonic acid from aqueous solution by composite resin.

Adsorption behavior of the iron impregnated, weakly basic resin D301 (Fe-D301) for removal of 2-naphthalenesulfonic acid (2-NSA) from aqueous solution was studied by using a fixed-bed column. The effects of process variables such as bed height, flow rate, and coexisting ions were investigated. The results indicated that the breakpoint and exhaustion point increased with increasing bed height and decreased with increasing 2-NSA flowrate. Experimental data showed a strong fit to the Bed Depth Service Time model. The coexisting ions in the 2-NSA solution had a clear effect on the breakthrough volume. The high extent of recovery of 2-NSA with good reproducibility provided an effective method for the separation of 2-NSA by the adsorbent Fe-D301.

Generation-dependent host-guest interactions: solution states of polyglycerol dendrimers of generations†3 and 4 modulate the localization of a guest molecule.

Host-guest interactions between polyglycerol dendrimers of generations 3 and 4 (PGD-G3 and G4) and 4-amino-3-hydroxynapthalene-2-sulphonic acid (AHSA) were investigated by fluorescence spectroscopy, isothermal titration calorimetry (ITC), and dynamic light scattering (DLS). PGD-G3 molecules were found to form an associated state with an average diameter of 82.7 nm in aqueous solution, in which PGD-G3 provided a much more polar microenvironment than glycerol. PGD-G3 and AHSA interacted attractively, showing a binding constant of 5.3×10(5)  M(-1) with a 2:1 stoichiometry. On the other hand, AHSA interacted with the periphery of PGD-G4, the majority of which existed as a unimer, forming a less polar microenvironment. The driving force of the interactions for PGD-G3 and -G4 were mainly enthalpically and entropically driven, respectively. The generation-dependent host-guest interactions were described in conjunction with thermodynamic parameters.

Degradation of reactive dyes in a photocatalytic circulating-bed biofilm reactor.

Decolorization and mineralization of reactive dyes by intimately coupled TiO2-photocatalysis and biodegradation (ICPB) on a novel TiO2-coated biofilm carrier were investigated in a photocatalytic circulating-bed biofilm reactor (PCBBR). Two typical reactive dyes--Reactive Black 5 (RB5) and Reactive Yellow 86 (RY86)--showed similar first-order kinetics when being photocatalytically decolorized at low pH (~4-5) in batch experiments. Photocatalytic decolorization was inhibited at neutral pH in the presence of phosphate or carbonate buffer, presumably due to electrostatic repulsion from negatively charged surface sites on TiO2, radical scavenging by phosphate or carbonate, or both. Therefore, continuous PCBBR experiments were carried out at a low pH (~4.5) to maintain high photocatalytic efficiency. In the PCBBR, photocatalysis alone with TiO2-coated carriers could remove target compound RB5 and COD by 97% and 47%, respectively. Addition of biofilm inside macroporous carriers maintained a similar RB5 removal efficiency, but COD removal increased to 65%, which is evidence of ICPB despite the low pH. ICPB was further proven by finding microorganisms inside carriers at the end of the PCBBR experiments. A proposed ICPB pathway for RB5 suggests that a major intermediate, a naphthol derivative, was responsible for most of the residual COD, while most of the nitrogen in the azo-bonds (-N=N-) was oxidized to N2.

Effect of peptide and guest charge on the structural, mechanical and release properties of ß-sheet forming peptides.

The effect of peptide charge on the self-assembly, gelation behavior, and model drug release profiles has been explored here for three octa-peptides, VEVKVEVK (VEK2), VKVKVEVK (VEK3), and VEVEVKVE (VEK1), that carry a net charge of 0, +2, and -2 at neutral pH, respectively. Transparent, self-supporting hydrogels were found to form above a critical concentration when the peptide charge modulus was >1 and this was independent of the sign of the charge. TEM, SAXS, and shear rheology revealed that there were no differences in hydrogel structure or mechanical properties when the peptides were at the same concentration and carried the same charge modulus. All peptides were found to form dense fibrillar networks formed by ß-sheet rich single fibers where lateral aggregation of the fibers occurred and increased with decreasing charge modulus. Such behavior was found to correlate with an increase in hydrogel mechanical properties, demonstrating that fiber lateral aggregation is inextricably linked with the mechanical properties of these hydrogels. Two hydrophilic model drug molecules, namely napthol yellow (NY) and martius yellow (MY), were subsequently incorporated within the VEK1 and VEK3 hydrogels at pH 7 and although they did not effect the self-assembly of the peptide at a molecular level, they did effect the level of lateral fiber aggregation observed and, therefore, the mechanical properties of the hydrogels. The release of each molecule from the hydrogels was monitored over time and shown to be controlled by Fickian diffusion where the diffusion rate, D, was dependent on the ratio between the overall effective charges carried by the peptide, i.e., the fibrillar network, and the overall charges carried by the guest molecules, but independent from the hydrogel concentration and mechanical properties within the ranges investigated. This work highlights the possibility of controlling the rate of release of small drug molecules by manipulating the charges on the guest molecules as well as the charged state of the self-assembling peptide.

Production of ligninolytic enzymes by solid-state fermentation using Pleurotus eryngii.

Pleurotus eryngii (DC.) Gillet (MCC58) was investigated for its ability to produce various ligninolytic enzymes such as laccase (Lac), manganese peroxidase (MnP), aryl alcohol oxidase (AAO), and lignin peroxidase (LiP) by solid-state fermentation (SSF), which was carried out using a support substrate from the fruit juice industry. The chemical content of grape waste from this industry was studied. Also, the production patterns of these extracellular enzymes were researched during the growth of the organism for a period of 20 days and the protein, reducing sugar, and nitrogen levels were monitored during the stationary cultivation. The highest Lac activity was obtained as 2247.62 ± 75 U/L on day 10 in the presence of 750 µM Mn²âº, while the highest MnP activity was attained as 2198.44 ± 65 U/L on day 15 in the presence of 500 µM Mn²âº. Decolorization of methyl orange and reactive red 2 azo dyes was also achieved with ligninolytic enzymes, produced in SSF of P. eryngii.

Ionization behavior of amino lipids for siRNA delivery: determination of ionization constants, SAR, and the impact of lipid pKa on cationic lipid-biomembrane interactions.

Ionizable amino lipids are being pursued as an important class of materials for delivering small interfering RNA (siRNA) therapeutics, and research is being conducted to elucidate the structure-activity relationships (SAR) of these lipids. The pK(a) of cationic lipid headgroups is one of the critical physiochemical properties of interest due to the strong impact of lipid ionization on the assembly and performance of these lipids. This research focused on developing approaches that permit the rapid determination of the relevant pK(a) of the ionizable amino lipids. Two distinct approaches were investigated: (1) potentiometric titration of amino lipids dissolved in neutral surfactant micelles; and (2) pH-dependent partitioning of a fluorescent dye to cationic liposomes formulated from amino lipids. Using the approaches developed here, the pK(a) values of cationic lipids with distinct headgroups were measured and found to be significantly lower than calculated values. It was also found that lipid-lipid interaction has a strong impact on the pK(a) values of lipids. Lysis of model biomembranes by cationic lipids was used to evaluate the impact of lipid pK(a) on the interaction between cationic lipids and cell membranes. It was found that cationic lipid-biomembrane interaction depends strongly on lipid pK(a) and solution pH, and this interaction is much stronger when amino lipids are highly charged. The presence of an optimal pK(a) range of ionizable amino lipids for siRNA delivery was suggested based on these results. The pK(a) methods reported here can be used to support the SAR screen of cationic lipids for siRNA delivery, and the information revealed through studying the impact of pK(a) on the interaction between cationic lipids and cell membranes will contribute significantly to the design of more efficient siRNA delivery vehicles.

Zeolite modification with cellulose nanofiber/magnetic nanoparticles for the elimination of reactive red 198.

In this paper for the first time, a cost-effective reinforced zeolite with cellulose nanofibers and magnetic nanoparticles (MZeo/Cellulose nanofiber) was used for the elimination of reactive red 198 (RR198) dye. The fabricated sorbent was characterized by SEM, FTIR, and XRD. The effect of operational parameters, including pH, RR198 concentration, the mass ratios of zeolite to cellulose nanofiber and zeolite coated cellulose to Fe3O4 nanoparticles, contact time, agitation speed, sorbent dosage, and temperature were studied. The prepared sorbent exhibited the maximum removal efficiency of 99% for RR198 removal at 30 °C. The presence of other dyes along with the target dye did not negatively affect the adsorption process and RR198 removal efficiency from actual water samples seemed satisfactory and rational. Equilibrium studies confirmed that both Langmuir and Freundlich models described the RR198 adsorption on MZeo/Cellulose nanofiber indicating physical and chemical interactions between the sorbent and RR198 molecules. Kinetic studies demonstrated that pseudo-second-order fitted best with experimental data. Also, thermodynamic studies showed the endothermic nature of the adsorption process. Compared to zeolite, MZeo/Cellulose nanofiber represented a promising removal efficiency for the elimination of RR198 dye from contaminated water.

A spot test for detection of cobalt release - early experience and findings.

BACKGROUND: It is often difficult to establish clinical relevance of metal exposure in cobalt-allergic patients. Dermatologists and patients may incorrectly assume that many metallic items release cobalt at levels that may cause cobalt dermatitis. Cobalt-allergic patients may be unaware that they are exposed to cobalt from handling work items, causing hand dermatitis. OBJECTIVES: To present early findings with a newly developed cobalt spot test. METHODS AND RESULTS: A cobalt spot test based on disodium-1-nitroso-2-naphthol-3,6-disulfonate was able to identify cobalt release at 8.3 ppm. The test may also be used as a gel test if combined with an agar preparation. We found no false-positive reactions when testing metals and alloys known not to contain cobalt. However, one cobalt-containing alloy, which elicited cobalt dermatitis in cobalt-allergic patients, was negative upon cobalt gel testing. CONCLUSIONS: The cobalt test detects amounts of cobalt release that approximate the elicitation concentration seen in cobalt-allergic patients. It may serve as a useful tool in dermatology offices and workplaces.

Enhanced degradation of Acid Red 73 by using cellulose-based hydrogel coated Fe3O4 nanocomposite as a Fenton-like catalyst.

Carboxymethyl cellulose-based hydrogel coated Fe3O4 magnetic nanoparticles were prepared using a coprecipitation combining graft copolymerization method, and characterized by various techniques to study their structure-property relationships. The nanocomposite was used as a heterogeneous Fenton-like catalyst for Acid Red 73 degradation. The effects of several key parameters, solution pH, H2O2 concentration, catalyst dosage, and temperature of the reaction medium on the pseudo-first-order kinetics of dye degradation was evaluated. The results showed that the nanocomposite catalyst were highly effective in activating H2O2 to produce reactive radicals for dye degradation, achieving complete decomposition under optimal conditions of 300 min at 25 °C and pH 3.5 with 100 mM H2O2 and 200 mg·L-1 catalyst. The complexing hydrogel-Fe2+/Fe3+ were the key factors that speed up the redox cycling between Fe2+ and Fe3+ species, thus accelerate the fast degradation rate of target pollutants. Scavenging experiments and electron paramagnetic resonance analyses revealed that Acid Red 73 was decomposed mainly by the attack of •OH radicals. Besides, reusability of the prepared nanocatalyst was also tested.

A naked-eye detection polyvinyl alcohol/cellulose-based pH sensor for intelligent packaging.

Naked-eye detection pH sensor is becoming a powerful tool in food safety monitoring. In this work, a pH sensor was developed by incorporating cellulose modified with acidochromic dye into polyvinyl alcohol (PVA). The results indicated that the dye (up to 170.4 µmol/g) was successfully anchored to cellulose. It was demonstrated that the addition of acidochromic regenerated cellulose (ARC) resulted in enhancement of tensile strength, elongation at break and maximum decomposition temperature by 44 %, 43.6 % and 11 °C, respectively. The pH sensor demonstrated that a visible color change from yellow to brick-red and to purple when placed in solutions of pH = 7, 10 and 12. The pH sensor showed excellent resistance to leaching under strong acidic and alkaline conditions. When applied to spoiled shrimp, an evident color change from yellow to brown was observed, suggesting it could serve as an easy-to-use, non-destructive visual indicator system for real-time food monitoring.

Electrostatic interactions regulate the release of small molecules from supramolecular hydrogels.

Supramolecular hydrogels have great potential as biomaterials for sustained delivery of therapeutics. While peptide-based supramolecular hydrogels have been developed that show promise for drug delivery applications, the high cost of production has limited their widespread adoption. Low molecular weight (LMW) supramolecular hydrogels are emerging as attractive and inexpensive alternatives to peptide-based hydrogels. We recently reported novel cationic fluorenylmethyloxycarbonyl-modified phenylalanine (Fmoc-Phe) hydrogels for localized and sustained in vivo release of an anti-inflammatory agent for functional pain remediation. In an effort to further elucidate design principles to optimize these materials for delivery of a variety of molecular agents, we herein report a systematic examination of electrostatic effects on the release of cargo molecules from Fmoc-Phe derived hydrogels. Specifically, we interrogate the release of cationic, anionic, and neutral cargo molecules from a series of cationic and anionic Fmoc-Phe derived hydrogels. We observed that cargo was readily released from the hydrogels except when the cargo and hydrogel network had complementary charges, in which case the cargo was highly retained in the network. These results demonstrate that the electrostatic characteristics of both the hydrogel network and the specific cargo are critical design parameters in the formulation of LMW supramolecular hydrogel systems in the development of next-generation materials for drug delivery applications.

Discerning Dynamic Signatures of Membrane-Bound α-Synuclein Using Site-Specific Fluorescence Depolarization Kinetics.

α-Synuclein is an intrinsically disordered protein that adopts an α-helical structure upon binding to the negatively charged lipid membrane. Binding-induced conformational change of α-synuclein plays a crucial role in the regulation of synaptic plasticity. In this work, we utilized the fluorescence depolarization kinetics methodology to gain the site-specific dynamical insights into the membrane-bound α-synuclein. We took advantage of the nonoccurrence of Cys in α-synuclein and created single-Cys variants at different sites for us to be able to label it with a thiol-active fluorophore. Our fluorescence depolarization results reveal the presence of three dynamically distinct types of motions of α-synuclein on POPG (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol)) small unilamellar vesicles (SUVs): (i) the (local) wobbling-in-cone motion of the fluorophore on the subnanosecond timescale, (ii) the backbone segmental mobility on the nanosecond timescale, and (iii) a slow depolarization component with a characteristic long rotational correlation time (∼60 ns) that is independent of the residue position. This characteristic timescale could potentially arise due to global tumbling of the protein-membrane complex, the global reorientation of only the protein within the membrane, and/or the translation diffusion of the protein on the curved membrane surface that could result in fluorescence depolarization due to the angular displacement of the transition dipole. In order to discern the molecular origin of the characteristic long rotational correlation time, we then carried our depolarization experiments varying the curvature of the membrane and varying the binding affinity by changing the lipid headgroup. These experiments revealed that the long rotational correlation time primarily arises due to the translational diffusion of α-synuclein on the curved membrane surface with a diffusion coefficient of ∼8.7 × 10-10 m2/s. The site-specific fluorescence depolarization methodology will find broad application in quantifying diffusion of a wide range of membrane-associated proteins involved in functions and diseases.