Freeze-drying of proteins with glass-forming oligosaccharide-derived sugar alcohols

Kadoya, Saori; Fujii, K.; Izutsu, Ken‐ichi; Yonemochi, Etsuo; Terada, Katsuhide; Yomota, Chikako; Kawanishi, Toru

doi:10.1016/j.ijpharm.2010.01.027

Cited by 62 publications

(33 citation statements)

References 44 publications

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“…The impossible access of polysaccharide to the CPO active site through the substrate channel due to steric hindrance made it impossible for polysaccharide to hold down the oxidation of the porphyrin ring. Moreover, if the second and third ways were true, the oxidation damage of the surrounding protein molecule should be inhibited by the additives because of the protection of polysaccharides forming a glass state around the enzyme molecule surfaces [26]; however, this result was not presented in our results. The conclusion that the main molecular event during the peroxide-mediated inactivation of chloroperoxidase was heme destruction has recently been testified [4].…”

Section: Effect Of Polysaccharides On Cpo Tolerance To Hydrogen Peroxidecontrasting

confidence: 58%

“…It is possible that the lowering of aggregation in the presence of sugars is, at least partly, due to an increase in the cohesive force of the medium and the lowering of protein diffusion. (3) Polysaccharides would further form a glass state around the enzyme molecules [26] (Scheme 1) to prevent the enzyme from losing activity caused by exposure to high temperatures.…”

Section: Stabilized Effect Of Polysaccharides On Cpo Activity At Elevmentioning

confidence: 99%

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Activity and Stability of Chloroperoxidase in the Presence of Small Quantities of Polysaccharides: A Catalytically Favorable Conformation Was Induced

Wang

Jiang

et al. 2011

Appl Biochem Biotechnol

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Chloroperoxidase (CPO) is thought to be the most versatile heme-containing enzyme with enormous applications in organic synthesis, biotransformation, pharmaceutical production, and detoxification of environmental pollutants. Any improvement in the stability of this enzyme will greatly enhance its application in the mentioned areas. In the present study, the effects of three polysaccharides (soluble starch, β-cyclodextrin, and dextrin) on the stability of CPO at elevated temperatures (20, 30, 35, 40, and 50 °C) or in aqueous-organic solvents media (methanol, dioxane, DMSO, and DMF) were investigated. An improved catalytic performance of CPO was observed in the presence of a small amount of the three polysaccharides, where dextrin provided the most effective promotion. The changes of enzyme structure and microenvironment around heme in the presence of additives were studied by fluorescence, circular dichroism, and UV-vis spectra analyses, as well as kinetic parameters measurement. A catalytically favorable structure of CPO was induced, including the strengthening of the α-helix structure and more exposure of heme for easy access of the substrate, resulting in an increase of catalytic turnover frequency (k (cat)) and the improvement of affinity and selectivity of CPO to substrate. The results revealed that the introduction of trace soluble starch, β-cyclodextrin, and dextrin (<10 μmol/L) in reaction media was an effective strategy for the enhancement of the thermodynamic and the operational stability of the enzyme, which are promising in view of the industrial applications of this versatile biological catalyst.

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Section: Effect Of Polysaccharides On Cpo Tolerance To Hydrogen Peroxidecontrasting

confidence: 58%

Section: Stabilized Effect Of Polysaccharides On Cpo Activity At Elevmentioning

confidence: 99%

Activity and Stability of Chloroperoxidase in the Presence of Small Quantities of Polysaccharides: A Catalytically Favorable Conformation Was Induced

Wang

Jiang

et al. 2011

Appl Biochem Biotechnol

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“…However, in order to ensure long term stability of protein pharmaceutical preparations, the moisture level shouldn't exceed 9% which is enough to J U S T A C C E P T E D hydrate the active site cleft of the proteins (16). Some excipients has cryoprotective and lyoprotective properties accordingly used to stabilise proteins in freeze drying process (see for examples; hydroxyethyl cellulose used with lactate dehydrogenase (17), polysorbate 20, trehalose, β-cyclodextrin and hydroxylethyl starch with glucagon (18), trehalose with insulin (19), pluronic F68 with calcitonin (20) and maltotriitol, trehalose, maltitol, and lactitol with L-lactate dehydrogenase and bovine serum albumin (21).…”

Section: Introductionmentioning

confidence: 99%

Influences of copolymers (Copovidone, Eudragit RL PO and Kollicoat MAE 30 DP) on stability and bioactivity of spray-dried and freeze-dried lysozyme

Haj-Ahmad

Mamayusupov

Elkordy

et al. 2016

Drug Development and Industrial Pharmacy

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Protein stability is the most crucial factor in protein pharmaceutical preparations. Various techniques were applied for producing stable protein formulations such as spray-drying and freeze-drying. However, heating and freezing stresses are disadvantages for proteins using these methods, respectively. Accordingly, excipients have been used to preserve therapeutic effects of proteins during processing and for long period of time. Therefore, influences of Copovidone, Eudragit® RL-PO and Kollicoat® MAE-30 DP (as excipients) on stability and integrity of lysozyme (as a model protein) in spray-dried and freeze-dried forms were investigated. Protein formulations in both dried forms were prepared without and with the addition of mentioned excipients at different concentrations. Protein formulations were characterised for yield determination, morphology using scanning electron microscopic (SEM), thermal analysis by Differential Scanning Calorimetry (DSC), secondary structure stability using Fourier transform infrared (FT-IR) spectroscopy and biological activity. All protein formulations were subjected to a stability study as solid protein formulations for 3 weeks at 24 °C/76% relative humidity and aqueous protein samples were stored at 50°C for 30 minutes in a water bath. Results showed that Copovidone successfully preserved integrity and biological activity of lysozyme before and after storage in both spray-dried and freezedried forms with more advantage for using higher concentration of the same excipient.Smooth spheres of spray-dried lysozyme formulations with Copovidone were smaller than spray-dried lysozyme without and with Kollicoat® MAE-30 DP, which affected %yield produced. Copovidone has demonstrated valuable protection ability for lysozyme.

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“…However, the most widely used techniques to dehydrate proteins are vacuum freeze drying (FD). The FD dehydration can minimize the losses of active components of foods owing to low temperature and low absolute pressure [2]. In the other hand, FD is also recognized as an expensive technology, because it requires much energy.…”

Section: Introductionmentioning

confidence: 99%

Dehydration of crude protein from Ginkgo biloba L. by microwave freeze drying

Fan

Ding

Liu

et al. 2012

International Journal of Biological Macromolecules

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Freeze-drying of proteins with glass-forming oligosaccharide-derived sugar alcohols

Cited by 62 publications

References 44 publications

Activity and Stability of Chloroperoxidase in the Presence of Small Quantities of Polysaccharides: A Catalytically Favorable Conformation Was Induced

Activity and Stability of Chloroperoxidase in the Presence of Small Quantities of Polysaccharides: A Catalytically Favorable Conformation Was Induced

Influences of copolymers (Copovidone, Eudragit RL PO and Kollicoat MAE 30 DP) on stability and bioactivity of spray-dried and freeze-dried lysozyme

Dehydration of crude protein from Ginkgo biloba L. by microwave freeze drying

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