The Effect of Ethylene Glycol, Glycine Betaine, and Urea on Lysozyme Thermal Stability

Schwinefus, Jeffrey J.; Leslie, Elizabeth J.; Nordstrom, Anna R.

doi:10.1021/ed100124n

Cited by 5 publications

(6 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Interestingly, it was observed that LYZ when stored in MEs shows high activity compared to the control sample employed in equal concentration. This observation may be attributed to the change in the 2° structure as evidenced by CD measurements, where content of β-sheet increases after heat treatment along with negligible change in microenvironment of Trp residues present in the active site . Another reason for such behavior could be the aggregation of LYZ in EG as also suggested by DLS measurements (Figure S11), when employed at the same concentration.…”

Section: Resultsmentioning

confidence: 75%

“…A decrease in emission intensity ( I em ) of the inherent fluorescence of LYZ with rise in temperature is observed, which is regained on cooling (Figures B and S8, Supporting Information). LYZ is reported to remain stable up to 70 °C in buffer and in EG, , and therefore such a decrease in I em is assigned to the polarity changes around hydrophobically buried Trp residues, which could be accompanied by 2° structural changes in LYZ. ,, Another reason for such decrease in I em may be an obvious decrease in viscosity of the system at higher temperatures.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Thermally Stable Ionic Liquid-Based Microemulsions for High-Temperature Stabilization of Lysozyme at Nanointerfaces

et al. 2019

View full text Add to dashboard Cite

The development of new strategies for thermal stability and storage of enzymes is very important, considering the nonretention of catalytic activity by enzymes under harsh conditions of temperature. Following this, herein, a new approach based on the interfacial adsorption of lysozyme (LYZ) at nanointerfaces of ionic liquid (IL)-based microemulsions, for enhanced thermal stability of LYZ, is reported. Microemulsions (MEs) composed of dialkyl imidazoliumbased surface active ILs (SAILs) as surfactants, ILs as the nonpolar phase, and ethylene glycol (EG) as the polar phase, without any cosurfactants, have been prepared and characterized in detail. Various regions corresponding to polar-in-IL, bicontinuous, and IL-in-polar phases have been characterized using conductivity measurements. Dynamic light scattering (DLS) measurements have provided insights into the size distribution of microdroplets, whereas temperature-dependent DLS measurements established the thermal stability of the MEs. Nanointerfaces formed by SAILs with EG in thermally stable reverse MEs act as fluid scaffolds to adsorb and provide thermal stability, up to 120 °C, to LYZ. Thermally treated LYZ upon extraction into a buffer shows enzyme activity owing to negligible change in the active site of LYZ, as marked by retention of microenvironment of Trp residues present in the active site of LYZ. The present work is expected to establish a new platform for the development of novel nanointerfaces utilizing biobased components for other biomedical applications.

show abstract

Section: Resultsmentioning

confidence: 75%

Section: Resultsmentioning

confidence: 99%

Thermally Stable Ionic Liquid-Based Microemulsions for High-Temperature Stabilization of Lysozyme at Nanointerfaces

et al. 2019

View full text Add to dashboard Cite

show abstract

“…The experiment described avoids the need for cell culture facilities and expensive ratiometric dyes commonly used to determine intracellular biochemical and clinical parameters, but provides students with the tools they will need to understand, apply, and develop ratiometric assays in their professional careers. Recently, in this Journal experiments were described where the thermal denaturation of lysozyme was followed by differential scanning calorimetry and UV-absorption spectroscopy. , …”

mentioning

confidence: 99%

Application of Ratiometric Measurements and Microplate Fluorimetry to Protein Denaturation: An Experiment for Analytical and Biochemistry Students

Marquês

Almeida

2013

J. Chem. Educ.

View full text Add to dashboard Cite

The number of applications of fluorescence spectroscopy in different areas of chemistry has increased dramatically, in part because a variety of instruments are used to measure fluorescence, including high-throughput microplate readers. Therefore, it is important to introduce students to different instruments. With many instruments, several experimental limitations hamper quantitative treatment of data, unless ratiometric measurements, that is, the ratio of intensity at two different excitation or emission wavelengths, are made. However, such methods are not always applicable. The denaturation of proteins often induces a red-shift of the tryptophan residues emission. Such a shift permits the use of ratiometric measures to obtain the fraction of native and denatured protein. To our knowledge, the use of ratiometric analysis with fluorescence measurements obtained from a microplate reader for the study of protein (biomolecular) denaturation has not been applied as a teaching exercise. In this experiment, the denaturation of hen egg-white lysozyme by guanidine hydrochloride is studied. Students perform ratiometric and single-wavelength measurements and obtain thermodynamic parameters for the denaturation process; they also test the reversibility of denaturation. In these studies the advantages of the ratiometric method are highlighted. Students develop analytical skills and, simultaneously, their understanding of the physical-chemical principles behind protein structural changes.

show abstract

“…For example, linking with other AFM labs [2,3,9,21] it can be part of a series on atomic force microscopy in advanced instrumental analysis/physical chemistry/biophysics/nanotechnology courses or be part of a series on protein structure and function in biochemistry/biotechnology/structural biology courses. Lysozyme is a frequently used model protein in the latter teaching labs [18,[22][23][24][25][26][27][28].…”

Section: Student Feedback Laboratory Time Management and Extensionmentioning

confidence: 99%

Laboratory exercise for studying the morphology of heat‐denatured and amyloid aggregates of lysozyme by atomic force microscopy

Gokalp

Horton

Jónsdóttir-Lewis

et al. 2017

Biochem Molecular Bio Educ

View full text Add to dashboard Cite

To facilitate learning advanced instrumental techniques, essential tools for visualizing biomaterials, a simple and versatile laboratory exercise demonstrating the use of Atomic Force Microscopy (AFM) in biomedical applications was developed. In this experiment, the morphology of heat-denatured and amyloid-type aggregates formed from a low-cost and well-characterized model protein, hen egg white lysozyme (HEWL), are compared. Structural differences between the amorphous and ordered particles are quantified using ImageJ for the analysis of AFM images as a postlaboratory assignment. The laboratory exercise allows the direct observation of changes in the protein structure and helps students to understand the operation of AFM, as well as protein folding and misfolding related to many physiological and pathological processes. The described protocol stands alone, but also fits well into a larger module on protein structure and function or microscopic techniques as it can be linked easily to existing laboratory exercises on these topics. It can be easily adapted to the upper level undergraduate laboratory courses with limited lab hours as well as graduate level courses to improve students' research skills. © 2017 by The International Union of Biochemistry and Molecular Biology, 46(2):162-171, 2018.

show abstract

The Effect of Ethylene Glycol, Glycine Betaine, and Urea on Lysozyme Thermal Stability

Cited by 5 publications

References 11 publications

Thermally Stable Ionic Liquid-Based Microemulsions for High-Temperature Stabilization of Lysozyme at Nanointerfaces

Thermally Stable Ionic Liquid-Based Microemulsions for High-Temperature Stabilization of Lysozyme at Nanointerfaces

Application of Ratiometric Measurements and Microplate Fluorimetry to Protein Denaturation: An Experiment for Analytical and Biochemistry Students

Laboratory exercise for studying the morphology of heat‐denatured and amyloid aggregates of lysozyme by atomic force microscopy

Contact Info

Product

Resources

About