The Impact of Bioconjugation on the Interfacial Activity of a Protein Biosurfactant

2018 ◽  
Author(s):  
Hossam H Tayeb ◽  
Marina Stienecker ◽  
Anton Middelberg ◽  
Frank Sainsbury
Keyword(s):  
Polyethylene Glycol ◽  
Colloidal Stability ◽  
Point Mutations ◽  
Pharmaceutical Formulations ◽  
Industrial Processes ◽  
Parent Molecule ◽  
Site Specific ◽  
Interfacial Activity ◽  
Surface Active ◽  
The Impact

Biosurfactants, are surface active molecules that can be produced by renewable, industrially scalable biologic processes. DAMP4, a designer biosurfactant, enables the modification of interfaces via genetic or chemical fusion to functional moieties. However, bioconjugation of addressable amines introduces heterogeneity that limits the precision of functionalization as well as the resolution of interfacial characterization. Here we designed DAMP4 variants with cysteine point mutations to allow for site-specific bioconjugation. The DAMP4 variants were shown to retain the structural stability and interfacial activity characteristic of the parent molecule, while permitting efficient and specific conjugation of polyethylene glycol (PEG). PEGylation results in a considerable reduction on the interfacial activity of both single and double mutants. Comparison of conjugates with one or two conjugation sites shows that both the number of conjugates as well as the mass of conjugated material impacts the interfacial activity of DAMP4. As a result, the ability of DAMP4 variants with multiple PEG conjugates to impart colloidal stability on peptide-stabilized emulsions is reduced. We suggest that this is due to constraints on the structure of amphiphilic helices at the interface. Specific and efficient bioconjugation permits the exploration and investigation of the interfacial properties of designer protein biosurfactants with molecular precision. Our findings should therefore inform the design and modification of biosurfactants for their increasing use in industrial processes, and nutritional and pharmaceutical formulations.

The Impact of Bioconjugation on the Interfacial Activity of a Protein Biosurfactant

2019 ◽  
Author(s):  
Hossam H Tayeb ◽  
Marina Stienecker ◽  
Anton Middelberg ◽  
Frank Sainsbury
Keyword(s):  
Colloidal Stability ◽  
Point Mutations ◽  
Pharmaceutical Formulations ◽  
Industrial Processes ◽  
Parent Molecule ◽  
Site Specific ◽  
Interfacial Activity ◽  
Surface Active ◽  
The Impact ◽  
Steric Constraints

Biosurfactants, are surface active molecules that can be produced by renewable, industrially scalable biologic processes. DAMP4, a designer biosurfactant, enables the modification of interfaces via genetic or chemical fusion to functional moieties. However, bioconjugation of addressable amines introduces heterogeneity that limits the precision of functionalization as well as the resolution of interfacial characterization. Here we designed DAMP4 variants with cysteine point mutations to allow for site-specific bioconjugation. The DAMP4 variants were shown to retain the structural stability and interfacial activity characteristic of the parent molecule, while permitting efficient and specific conjugation of polyethylene glycol (PEG). PEGylation results in a considerable reduction on the interfacial activity of both single and double mutants. Comparison of conjugates with one or two conjugation sites shows that both the number of conjugates as well as the mass of conjugated material impacts the interfacial activity of DAMP4. As a result, the ability of DAMP4 variants with multiple PEG conjugates to impart colloidal stability on peptide-stabilized emulsions is reduced. We suggest that this is due to steric constraints on the structure of amphiphilic helices at the interface. Specific and efficient bioconjugation permits the exploration and investigation of the interfacial properties of designer protein biosurfactants with molecular precision. Our findings should therefore inform the design and modification of biosurfactants for their increasing use in industrial processes, and nutritional and pharmaceutical formulations.

The Impact of Bioconjugation on the Interfacial Activity of a Protein Biosurfactant

2019 ◽  
Author(s):  
Hossam H Tayeb ◽  
Marina Stienecker ◽  
Anton Middelberg ◽  
Frank Sainsbury
Keyword(s):  
Colloidal Stability ◽  
Point Mutations ◽  
Pharmaceutical Formulations ◽  
Industrial Processes ◽  
Parent Molecule ◽  
Site Specific ◽  
Interfacial Activity ◽  
Surface Active ◽  
The Impact ◽  
Steric Constraints

Biosurfactants, are surface active molecules that can be produced by renewable, industrially scalable biologic processes. DAMP4, a designer biosurfactant, enables the modification of interfaces via genetic or chemical fusion to functional moieties. However, bioconjugation of addressable amines introduces heterogeneity that limits the precision of functionalization as well as the resolution of interfacial characterization. Here we designed DAMP4 variants with cysteine point mutations to allow for site-specific bioconjugation. The DAMP4 variants were shown to retain the structural stability and interfacial activity characteristic of the parent molecule, while permitting efficient and specific conjugation of polyethylene glycol (PEG). PEGylation results in a considerable reduction on the interfacial activity of both single and double mutants. Comparison of conjugates with one or two conjugation sites shows that both the number of conjugates as well as the mass of conjugated material impacts the interfacial activity of DAMP4. As a result, the ability of DAMP4 variants with multiple PEG conjugates to impart colloidal stability on peptide-stabilized emulsions is reduced. We suggest that this is due to steric constraints on the structure of amphiphilic helices at the interface. Specific and efficient bioconjugation permits the exploration and investigation of the interfacial properties of designer protein biosurfactants with molecular precision. Our findings should therefore inform the design and modification of biosurfactants for their increasing use in industrial processes, and nutritional and pharmaceutical formulations.

scholarly journals TransfersomILs: From Ionic Liquids to a New Class of Nanovesicular Systems

Nanomaterials ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 7
Author(s):  
Ana Júlio ◽  
João Guilherme Costa ◽  
Catarina Pereira-Leite ◽  
Tânia Santos de Almeida
Keyword(s):  
Ionic Liquids ◽  
Physicochemical Properties ◽  
Colloidal Stability ◽  
Storage Stability ◽  
Pharmaceutical Formulations ◽  
Controlled Delivery ◽  
New Class ◽  
Improved Performance ◽  
And Performance ◽  
The Impact

Ionic liquids (ILs) have increasingly been studied as key materials to upgrade the performance of many pharmaceutical formulations. In controlled delivery systems, ILs have improved multiple physicochemical properties, showing the relevance of continuing to study their incorporation into these formulations. Transfersomes are biocompatible nanovesicular systems, quite useful in controlled delivery. They have promising characteristics, such as elasticity and deformability, making them suitable for cutaneous delivery. Nonetheless, their overall properties and performance may still be improved. Herein, new TransfersomILs systems to load rutin were developed and the physicochemical properties of the formulations were assessed. These systems were prepared based on an optimized formulation obtained from a Box–Behnken factorial design (BBD). The impact of imidazole-based ILs, cholinium-based ILs, and their combinations on the cell viability of HaCaT cells and on the solubility of rutin was initially assessed. The newly developed TransfersomILs containing rutin presented a smaller size and, in general, a higher association efficiency, loading capacity, and total amount of drug release compared to the formulation without IL. The ILs also promoted the colloidal stability of the vesicles, upgrading storage stability. Thus, ILs were a bridge to develop new TransfersomILs systems with an overall improved performance.

Seed-mediated synthesis and PEG coating of gold nanoparticles for controlling morphology and sizes

MRS Advances ◽  
2020 ◽  
Vol 5 (63) ◽  
pp. 3353-3360
Author(s):  
Susana Helena Arellano Ramírez ◽  
Perla García Casillas ◽  
Christian Chapa González
Keyword(s):  
Gold Nanoparticles ◽  
Polyethylene Glycol ◽  
Room Temperature ◽  
Colloidal Stability ◽  
Infrared Spectrometry ◽  
Ammonium Bromide ◽  
Chloroauric Acid ◽  
Vis Spectroscopy ◽  
Seed Mediated ◽  
Peg Coating

AbstractA significant area of research is biomedical applications of nanoparticles which involves efforts to control the physicochemical properties through simple and scalable processes. Gold nanoparticles have received considerable attention due to their unique properties that they exhibit based on their morphology. Gold nanospheres (AuNSs) and nanorods (AuNRs) were prepared with a seed-mediated method followed of polyethylene glycol (PEG)-coating. The seeds were prepared with 0.1 M cetyltrimethyl-ammonium bromide (CTAB), 0.005 M chloroauric acid (HAuCl4), and 0.01 M sodium borohydride (NaBH4) solution. Gold nanoparticles with spherical morphology was achieved by growth by aggregation at room temperature, while to achieve the rod morphology 0.1 M silver nitrate (AgNO3) and 0.1 M ascorbic acid solution were added. The gold nanoparticles obtained by the seed-mediated synthesis have spherical or rod shapes, depending on the experimental conditions, and a uniform particle size. Surface functionalization was developed using polyethylene glycol. Morphology, and size distribution of AuNPs were evaluated by Field Emission Scanning Electron Microscopy. The average size of AuNSs, and AuNRs was 7.85nm and 7.96 x 31.47nm respectively. Fourier transform infrared spectrometry was performed to corroborate the presence of PEG in the AuNPs surface. Additionally, suspensions of AuNSs and AuNRs were evaluated by UV-Vis spectroscopy. Gold nanoparticles were stored for several days at room temperature and it was observed that the colloidal stability increased once gold nanoparticles were coated with PEG due to the shield formed in the surface of the NPs and the increase in size which were 9.65±1.90 nm of diameter for AuNSs and for AuNRs were 29.03±5.88 and 8.39±1.02 nm for length and transverse axis, respectively.

scholarly journals Modified Gold Nanoparticles for Efficient Delivery of Betulinic Acid to Cancer Cell Mitochondria

2021 ◽  
Vol 22 (10) ◽  
pp. 5072
Author(s):  
Olakunle Oladimeji ◽  
Jude Akinyelu ◽  
Aliscia Daniels ◽  
Moganavelli Singh
Keyword(s):  
Polyethylene Glycol ◽  
Cancer Cell ◽  
Betulinic Acid ◽  
Targeted Delivery ◽  
Epigallocatechin Gallate ◽  
High Amplitude ◽  
Significant Inhibition ◽  
Mitochondrial Targeting ◽  
The Impact

Advances in nanomedicine have seen the adaptation of nanoparticles (NPs) for subcellular delivery for enhanced therapeutic impact and reduced side effects. The pivotal role of the mitochondria in apoptosis and their potential as a target in cancers enables selective induction of cancer cell death. In this study, we examined the mitochondrial targeted delivery of betulinic acid (BA) by the mitochondriotropic TPP+-functionalized epigallocatechin gallate (EGCG)-capped gold NPs (AuNPs), comparing the impact of polyethylene glycol (PEG) and poly-L-lysine-graft-polyethylene glycol (PLL-g-PEG) copolymer on delivery efficacy. This included the assessment of their cellular uptake, mitochondrial localization and efficacy as therapeutic delivery platforms for BA in the human Caco-2, HeLa and MCF-7 cancer cell lines. These mitochondrial-targeted nanocomplexes demonstrated significant inhibition of cancer cell growth, with targeted nanocomplexes recording IC50 values in the range of 3.12–13.2 µM compared to that of the free BA (9.74–36.31 µM) in vitro, demonstrating the merit of mitochondrial targeting. Their mechanisms of action implicated high amplitude mitochondrial depolarization, caspases 3/7 activation, with an associated arrest at the G0/G1 phase of the cell cycle. This nano-delivery system is a potentially viable platform for mitochondrial-targeted delivery of BA and highlights mitochondrial targeting as an option in cancer therapy.

scholarly journals Ice Coverage Induced by Depositing a Water Drop onto the Supercooled Substrate at Extreme Low Vapor Pressure

Crystals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 691
Author(s):  
Yugang Zhao ◽  
Zichao Zuo ◽  
Haibo Tang ◽  
Xin Zhang
Keyword(s):  
Vapor Pressure ◽  
Substrate Temperature ◽  
Water Drop ◽  
Industrial Processes ◽  
Aircraft Icing ◽  
Ice Coverage ◽  
Fundamental Understanding ◽  
Order Of Magnitude ◽  
The Impact ◽  
Kinetics Of

Icing/snowing/frosting is ubiquitous in nature and industrial processes, and the accretion of ice mostly leads to catastrophic consequences. The existing understanding of icing is still limited, particularly for aircraft icing, where direct observation of the freezing dynamics is inaccessible. In this work, we investigate experimentally the impact and freezing of a water drop onto the supercooled substrate at extremely low vapor pressure, to mimic an aircraft passing through clouds at a relatively high altitude, engendering icing upon collisions with pendant drops. Special attention is focused on the ice coverage induced by an impinging drop, from the perimeter pointing outward along the radial direction. We observed two freezing regimes: (I) spread-recoil-freeze at the substrate temperature of Ts = −15.4 ± 0.2 °C and (II) spread (incomplete)-freeze at the substrate temperature of Ts = −22.1 ± 0.2 °C. The ice coverage is approximately one order of magnitude larger than the frozen drop itself, and counterintuitively, larger supercooling yields smaller ice coverage in the range of interest. We attribute the variation of ice coverage to the kinetics of vapor diffusion in the two regimes. This fundamental understanding benefits the design of new anti-icing technologies for aircraft.

scholarly journals Site-specific N-glycosylation analysis of animal cell culture-derived Zika virus proteins

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alexander Pralow ◽  
Alexander Nikolay ◽  
Arnaud Leon ◽  
Yvonne Genzel ◽  
Erdmann Rapp ◽  
...  
Keyword(s):  
Zika Virus ◽  
Animal Cell ◽  
Gradient Centrifugation ◽  
Viral Envelope ◽  
Protein Glycosylation ◽  
High Yield ◽  
Structural Protein ◽  
Site Specific ◽  
E Protein ◽  
The Impact

AbstractHere, we present for the first time, a site-specific N-glycosylation analysis of proteins from a Brazilian Zika virus (ZIKV) strain. The virus was propagated with high yield in an embryo-derived stem cell line (EB66, Valneva SE), and concentrated by g-force step-gradient centrifugation. Subsequently, the sample was proteolytically digested with different enzymes, measured via a LC–MS/MS-based workflow, and analyzed in a semi-automated way using the in-house developed glyXtoolMS software. The viral non-structural protein 1 (NS1) was glycosylated exclusively with high-mannose structures on both potential N-glycosylation sites. In case of the viral envelope (E) protein, no specific N-glycans could be identified with this method. Nevertheless, N-glycosylation could be proved by enzymatic de-N-glycosylation with PNGase F, resulting in a strong MS-signal of the former glycopeptide with deamidated asparagine at the potential N-glycosylation site N444. This confirmed that this site of the ZIKV E protein is highly N-glycosylated but with very high micro-heterogeneity. Our study clearly demonstrates the progress made towards site-specific N-glycosylation analysis of viral proteins, i.e. for Brazilian ZIKV. It allows to better characterize viral isolates, and to monitor glycosylation of major antigens. The method established can be applied for detailed studies regarding the impact of protein glycosylation on antigenicity and human pathogenicity of many viruses including influenza virus, HIV and corona virus.

The impact of acid hydrolysis conditions on carbohydrate determination in lignocellulosic materials: a case study with Eucalyptus globulus bark

Holzforschung ◽  
2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Ricardo Jorge Oliveira ◽  
Bruna Santos ◽  
Maria J. Mota ◽  
Susana R. Pereira ◽  
Pedro C. Branco ◽  
...  
Keyword(s):  
Acid Hydrolysis ◽  
Eucalyptus Globulus ◽  
Industrial Processes ◽  
Lignocellulosic Materials ◽  
Optimal Conditions ◽  
Hydrolysis Conditions ◽  
The Impact ◽  
Eucalyptus Bark

Abstract Lignocellulosic biomass represents a suitable feedstock for production of biofuels and bioproducts. Its chemical composition depends on many aspects (e.g. plant source, pre-processing) and it has impact on productivity of industrial bioprocesses. Numerous methodologies can be applied for biomass characterisation, with acid hydrolysis being a particularly relevant step. This study intended to assess the most suitable procedures for acid hydrolysis, taking Eucalyptus globulus bark as a case study. For that purpose, variation of temperature (90–120 °C) was evaluated over time (0–5 h), through monosaccharides and oligosaccharides contents and degradation. For glucose, the optimal conditions were 100 °C for 2.5 h, reaching a content of 48.6 wt.%. For xylose, the highest content (15.2 wt.%) was achieved at 90 °C for 2 h, or 120 °C for 0.5 h. Maximum concentrations of mannose and galactose (1.0 and 1.7 wt.%, respectively) were achieved at 90 and 100 °C (2–3.5 h) or at 120 °C (0.5–1 h). These results revealed that different hydrolysis conditions should be applied for different sugars. Using this approach, total sugar quantification in eucalyptus bark was increased by 4.3%, which would represent a 5% increase in the ethanol volume produced, considering a hypothetical bioethanol production yield. This reflects the importance of feedstock characterization on determination of economic viability of industrial processes.

scholarly journals The influence of riparian woodland on the spatial and temporal variability of stream water temperatures in an upland salmon stream

2004 ◽  
Vol 8 (3) ◽  
pp. 449-459 ◽  
Author(s):  
I. A. Malcolm ◽  
D. M. Hannah ◽  
M. J. Donaghy ◽  
C. Soulsby ◽  
A. F. Youngson
Keyword(s):  
Thermal Regime ◽  
Temporal Variability ◽  
Heat Budget ◽  
Riparian Forest ◽  
Stream Water ◽  
Spatial And Temporal Variability ◽  
Substantial Impact ◽  
Site Specific ◽  
The Impact ◽  
Riparian Woodland

Abstract. The spatio-temporal variability of stream water temperatures was investigated at six locations on the Girnock Burn (30km2 catchment), Cairngorms, Scotland over three hydrological years between 1998 and 2002. The key site-specific factors affecting the hydrology and climatology of the sampling points were investigated as a basis for physical process inference. Particular emphasis was placed on assessing the effects of riparian forest in the lower catchment versus the heather moorland riparian zones that are spatially dominant in the upper catchment. The findings were related to river heat budget studies that provided process detail. Gross changes in stream temperature were affected by the annual cycle of incoming solar radiation and seasonal changes in hydrological and climatological conditions. Inter-annual variation in these controlling variables resulted in inter-annual variability in thermal regime. However, more subtle inter-site differences reflected the impact of site-specific characteristics on various components of the river energy budget. Inter-site variability was most apparent at shorter time scales, during the summer months and for higher stream temperatures. Riparian woodland in the lower catchment had a substantial impact on thermal regime, reducing diel variability (over a period of 24 hours) and temperature extremes. Observed inter-site differences are likely to have a substantial effect on freshwater ecology in general and salmonid fish in particular. Keywords: temperature, thermal regime, forest, salmon, hydrology, Girnock Burn, Cairngorm

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