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.

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.

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.

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

scholarly journals In Planta Mutagenesis Determines the Functional Regions of the Wheat Puroindoline Proteins

Genetics ◽  
2009 ◽  
Vol 183 (3) ◽  
pp. 853-860 ◽  
Author(s):  
Leila Feiz ◽  
Brian S. Beecher ◽  
John M. Martin ◽  
Michael J. Giroux
Keyword(s):  
Functional Analysis ◽  
Allelic Variation ◽  
Screening Method ◽  
Point Mutations ◽  
Missense Mutations ◽  
Grain Hardness ◽  
In Planta ◽  
Rich Region ◽  
Critical Function ◽  
The Impact

In planta analysis of protein function in a crop plant could lead to improvements in understanding protein structure/function relationships as well as selective agronomic or end product quality improvements. The requirements for successful in planta analysis are a high mutation rate, an efficient screening method, and a trait with high heritability. Two ideal targets for functional analysis are the Puroindoline a and Puroindoline b (Pina and Pinb, respectively) genes, which together compose the wheat (Triticum aestivum L.) Ha locus that controls grain texture and many wheat end-use properties. Puroindolines (PINs) together impart soft texture, and mutations in either PIN result in hard seed texture. Studies of the PINs' mode of action are limited by low allelic variation. To create new Pin alleles and identify critical function-determining regions, Pin point mutations were created in planta via EMS treatment of a soft wheat. Grain hardness of 46 unique PIN missense alleles was then measured using segregating F2:F3 populations. The impact of individual missense alleles upon PIN function, as measured by grain hardness, ranged from neutral (74%) to intermediate to function abolishing. The percentage of function-abolishing mutations among mutations occurring in both PINA and PINB was higher for PINB, indicating that PINB is more critical to overall Ha function. This is contrary to expectations in that PINB is not as well conserved as PINA. All function-abolishing mutations resulted from structure-disrupting mutations or from missense mutations occurring near the Tryptophan-rich region. This study demonstrates the feasibility of in planta functional analysis of wheat proteins and that the Tryptophan-rich region is the most important region of both PINA and PINB.

scholarly journals (Re)discovering Toronto's waterfront: infrastructure and connectivity in a post-industrial landscape

2021 ◽  
Author(s):  
Geordie Gordon
Keyword(s):  
Industrial Processes ◽  
Urban Forms ◽  
Industrial Landscape ◽  
Course Of Action ◽  
Waterfront Redevelopment ◽  
Industrial Use ◽  
Post Industrial ◽  
The Impact ◽  
American Cities ◽  
The City

The transition of waterfront land use from industrial to post-industrial is a global phenomenon. There are several forces that are driving this change, including the advancement of shipping technology and the relocation of industrial processes to areas with greater availability of land. In place of industrial uses, many cities have undertaken, or are in the process of undertaking the redevelopment of their waterfront. As a result of past industrial use, there often exists, a significant amount of transportation infrastructure that isolates the city from the waterfront. This paper establishes the context for waterfront redevelopment, before examining the impact of infrastructure urban forms by using the work of Kevin Lynch as a tool for analysis. Several case precedents are used to examine the course of action that other North American cities have pursued to mitigate the impact of infrastructure forms on the waterfront and how they may influence the way Toronto deals with its waterfront infrastructure.

scholarly journals Assembly: Performing the materiality of Muslim prayer spaces

Scene ◽  
2018 ◽  
Vol 6 (2) ◽  
pp. 133-151 ◽  
Author(s):  
Julie Marsh
Keyword(s):  
Social Boundaries ◽  
Research Project ◽  
Site Specific ◽  
Specific Research ◽  
A Site ◽  
The Impact

This article reflects on the significance and impact of Assembly, a site-specific research project made and exhibited in Birmingham Central Mosque, Brick Lane Mosque and Old Kent Road Mosque from 2016 to 2020. Assembly provided an opportunity for Muslims and non-Muslims to experience Jumu’ah prayer first-hand via the site performances, which temporarily dissolved the religious/social boundaries of each mosque. Each performance highlighted the differences and relations between each site, furthering ideas of performativity in Muslim prayer spaces. This article summarizes the impact reported by each mosque community as well as reflecting upon the relationships built within the wider community.

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