Effects of trehalose and sorbitol on the activity and structure of Pseudomonas cepacia lipase: Spectroscopic insight

Azizi, Azadeh; Ranjbar, Bijan; Khajeh, Khosro; Ghodselahi, T.; Hoornam, Soraya; Mobasheri, Hamid; Ganjalikhany, Mohamad Reza

doi:10.1016/j.ijbiomac.2011.06.025

Cited by 28 publications

(27 citation statements)

References 36 publications

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“…This was proposed for the effect of increased colloidal stability on lysozyme in the presence of DMSO and glycerol 48 but was not found to be the case for Pseudomonas cepacia lipase in the presence of trehalose. 49 At pH 4.6 with 5% (w/v) NaCl, the electrostatic charge is screened and the net interaction potential is attractive. The addition of sugar had little effect on the colloidal stability of lysozyme.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Thermal and Solution Stability of Lysozyme in the Presence of Sucrose, Glucose, and Trehalose

James

McManus

2012

J. Phys. Chem. B

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The effect of the sugars sucrose, glucose, and trehalose on the structural and colloidal stability of lysozyme has been investigated using differential scanning calorimetry and quasi-elastic light scattering, respectively. While sugars are known to increase the temperature at which thermal denaturation of protein occurs, it is not clear if, under the same solution conditions, greater colloidal stability is achieved. The measurements were carried out on lysozyme in three different buffer solutions, 0.05 M sodium acetate (pH 4.6), 0.05 M sodium acetate with 5% (w/v) NaCl, and 10 mM sodium phosphate (pH 7.0). The results show that enhancement of structural stability in the presence of sugars is pH, salt concentration, and sugar dependent. Enhancement of colloidal stability in the presence of sugars, while also pH and salt concentration dependent, as expected, only correlates with increases in the structural stability when the solution behavior is not dominated by highly stabilizing electrostatic repulsive interactions. ■ INTRODUCTIONSugars are widely used excipients in the formulation of biotherapeutic products.1−7 Therapeutic proteins are produced and used for the treatment of various human diseases 8 such as insulin for the treatment of diabetes, 9 monoclonal antibodies for rheumatoid arthritis, 10 interferon-α for leukemia, 11 and interferon-β for multiple sclerosis.12 Sugars are generally used as protein structure stabilizers, 2,13 since additional stability is conferred to a protein during lyophilization by the addition of the sugar.14 Two hypotheses have been proposed to describe the stabilizing effect of sugars on proteins during lyophilization; one such hypothesis states that sugar acts as a water substituent and stabilizes proteins by forming hydrogen bonds at specific sites on the surface of the protein.14−16 Another hypothesis, referred to as the "vitrification hypothesis", states that disaccharides form sugar glasses, thereby immobilizing the protein molecules and providing protection against destabilizing reactions. 15,16 Lysozyme is a globular protein with a molecular weight of approximately 14.7 kDa. Lysozyme has been used as a model protein for this study, since its biophysical properties are well understood and the specific contribution of the sugar to its stability over a range of solution conditions can easily be compared with that of the native protein. Lysozyme is a widely studied protein due to its rich phase behavior 17,18 including crystallization, 19 liquid−liquid phase separation, 20−22 and the formation of equilibrium clusters 23−25 and gels. 24 The solution behavior of lysozyme is also well understood in terms of its interaction with salt ions. 26−28Protein stability by itself is a generic term, and it can be thought of in several ways. The stabilization of protein structure against thermal denaturation is referred to as its thermal or structural stability. 37 and presence of excipients. 6 At pH values away from the isoelectric point of the protein, there is an increase in cha...

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Section: ■ Results and Discussionmentioning

confidence: 99%

Thermal and Solution Stability of Lysozyme in the Presence of Sucrose, Glucose, and Trehalose

James

McManus

2012

J. Phys. Chem. B

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“…Circular dichroism spectropolarimeter is one of the best techniques to detect any conformational changes that might be induced in the structure of macromolecules [19,20]. To investigate the alterations of the secondary structure of antibody after conjugation with gold nanoparticles, CD spectra were taken by a J-810 spectropolarimeter (Jasco, Tokyo, Japan).…”

Section: Far-uv Circular Dichroism Studiesmentioning

confidence: 99%

“…Upon the immediate addition of toxoid, these molecules interact with the antibody at the surface of GNPs and the SPR shifts would be observed due to changes in the refractive index at the surface of the GNPs. In fact, the resonance wavelength of the surface plasmon of GNPs is highly dependent on the environment and the mass at the surface of the particles [20,27].…”

Section: Characterization Of Gold Nanoparticlesmentioning

confidence: 99%

Localized Surface Plasmon Resonance (LSPR) Detection of Diphtheria Toxoid Using Gold Nanoparticle-Monoclonal Antibody Conjugates

et al. 2017

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Detection of diphtheria toxin (DT) which is produced by Corynebacterium diphtheria, a zoonotic pathogen and a leading cause of diphtheria, is the critical step in the clinical laboratory. Traditional methods for DT detection are time consuming with low sensitivity. Herein, a localized surface plasmon resonance (LSPR) nanobioprobe has been developed based on specific immunological interactions between gold nanoparticles (GNPs) conjugated with monoclonal antibody and diphtheria toxoid in order for the rapid detection of DT. For this, plasmonic GNPs were conjugated to monoclonal antibodies covalently. The covalent conjugation has been confirmed by dynamic light scattering (DLS) and electrochemical techniques. Then, structural alterations of the conjugated antibody were monitored by circular dichroism (CD) and fluorescence spectroscopy methods. After that, the sensitivity of the nanobioprobe has been investigated via measuring the LSPR band λmax shifts of GNPs and LSPR sensitivity of nanobioprobe was compared with the ELISA method. Results suggested that this assay is highly selective and sensitive with a lower detection limit of about 10 ng/mL. The LSPR biosensor reduced the DT detection time from 2 or 3 days to less than 1 h compared with traditional methods. In conclusion, the investigation presents a rapid, sensitive, and selective method for the diagnosis of DT in clinical specimens.

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“…The strategy of feeding mixed substrates has been applied in other fermentations. This was an effective method to decrease cell mortality, maintain the enzyme activity, and promote targeted metabolite production [9][10][11]. Similar studies with regard to glutamate fermentation are also important, but few have been reported.…”

mentioning

confidence: 96%

“…These auxiliary substrates are usually non-repressive carbon sources. They can protect cells against stress in their living environment, enhance cell viability, and reduce the metabolic burden [9][10][11][12]. It was reported that the production of alkaline polygalacturonate lyase and lipase increased by 1.85-fold and 8.7-fold, respectively, when the strategy of methanol/sorbitol co-feeding was adopted [10,13].…”

mentioning

confidence: 99%

Improving the stability of glutamate fermentation by Corynebacterium glutamicum via supplementing sorbitol or glycerol

Cao

Shi

et al. 2015

Bioresour. Bioprocess.

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Background: Corynebacterium glutamicum is widely used in glutamate fermentation. The fermentation feature of the strain varies sometimes. These variations may lead to the reduction in the ability of the strain to resist environmental changes and to synthesize glutamate, resulting in abnormal glutamate fermentations. Results: In the abnormal glutamate fermentations, glutamate accumulation stopped after glucose feeding and the final glutamate concentration was at a lower level (50 to 60 g/L). The r NAD

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Effects of trehalose and sorbitol on the activity and structure of Pseudomonas cepacia lipase: Spectroscopic insight

Cited by 28 publications

References 36 publications

Thermal and Solution Stability of Lysozyme in the Presence of Sucrose, Glucose, and Trehalose

Thermal and Solution Stability of Lysozyme in the Presence of Sucrose, Glucose, and Trehalose

Localized Surface Plasmon Resonance (LSPR) Detection of Diphtheria Toxoid Using Gold Nanoparticle-Monoclonal Antibody Conjugates

Improving the stability of glutamate fermentation by Corynebacterium glutamicum via supplementing sorbitol or glycerol

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