How CTAB assists the refolding of native and recombinant lysozyme

Wang, Jun; Lu, Diannan; Lin, Ying; Liu, Zheng

doi:10.1016/j.bej.2005.03.004

Cited by 37 publications

(22 citation statements)

References 31 publications

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“…Refolding of denatured/reduced lysozyme by AMC-IEC Denatured/reduced lysozyme is widely used as a model protein to study a new refolding method, and it was previously refolded by several groups using IEC 24,30,31 and using AMC 32,33 separately. In this work, AMC and IEC were incorporated to form a new refolding method, AMC-IEC, and it was used to the refolding of urea-denatured/DTTreduced lysozyme.…”

Section: Resultsmentioning

confidence: 99%

Refolding of denatured/reduced lysozyme at high concentrations by artificial molecular chaperone‐ion exchange chromatography

Wang

Zhang

Chen

et al. 2010

Biotechnology Progress

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Development of high efficiency and low cost protein refolding methods is a highlighted research focus in biotechnology. Artificial molecular chaperone (AMC) and protein folding liquid chromatography (PFLC) are two attractive refolding methods developed in recent years. In the present work, AMC and one branch of PFLC, ion exchange chromatography (IEC), are integrated to form a new refolding method, artificial molecular chaperone-ion exchange chromatography (AMC-IEC). This new method is applied to the refolding of a widely used model protein, urea-denatured/dithiothreitol-reduced lysozyme. Many factors influencing the refolding of lysozyme, such as urea concentration, beta-cyclodextrin concentration, molar ratio of detergent to protein, mobile phase flow rate, and type of detergent, were investigated, respectively, to optimize the conditions for lysozyme refolding by AMC-IEC. Compared with normal IEC refolding method, the activity recoveries of lysozyme obtained by AMC-IEC were much higher in the investigated range of initial protein concentrations. Moreover, the activity recoveries obtained by using this newly developed refolding method were still quite high for denatured/reduced lysozyme at high initial concentrations. When the initial protein concentration was 200 mg mL(-1), the activity recovery was over 60%. In addition, the lifetime of the chromatographic column during AMC-IEC was much longer than that during protein refolding by normal IEC. Therefore, AMC-IEC is a high efficient and low cost protein refolding method.

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Section: Resultsmentioning

confidence: 99%

Refolding of denatured/reduced lysozyme at high concentrations by artificial molecular chaperone‐ion exchange chromatography

Wang

Zhang

Chen

et al. 2010

Biotechnology Progress

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“…Taking into consideration the chemical structure of ionic liquids, hydrophobic cations with longer alkyl chains are similar to cationic detergents. Cationic detergents such as cetyltrimethylammonium bromide suppress protein aggregation in protein refolding processes due to the hydrophobic interaction between the detergent molecules and the refolding intermediates [33,34]. Therefore, hydrophobic cations might also bind to the hydrophobic surfaces of proteins, thereby leading to the prevention of refolding and thus destabilization of the native conformation.…”

Section: Thermodynamic and Kinetic Stability Of Native Lysozyme In Thmentioning

confidence: 99%

Protein Refolding by N-Alkylpyridinium and N-Alkyl-N-methylpyrrolidinium Ionic Liquids

Yamamoto

Yamaguchi

Nagamune

2011

Appl Biochem Biotechnol

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An important property of ionic liquids consisting of cations and anions is that the chemical structures can be easily tuned. To expand the repertoire of effective ionic liquid-based refolding additives, we focused on this tunable property and investigated the effects of new candidates such as N-alkylpyridinium chlorides and N-alkyl-N-methylpyrrolidinium chlorides on protein refolding. Denatured lysozyme (30 mg/mL) was used as a model protein and refolded by 30-fold dilution with various refolding buffers containing different ionic liquids consisting of a systematic variety of alkyl chains. Compared with the refolding yield without additives (lower than 10%), less hydrophobic ionic liquids such as N-ethyl, N-butyl and N-hexylpyridinium chlorides, and N-butyl-N-methylpyrrolidinium chloride were effective in enhancing the refolding yields (46-69%), because they primarily suppressed aggregation because of their chaotropic properties. N-alkylpyridinium cations were more hydrophobic than N-alkyl-N-methylpyrrolidinium cations according to the calculated log P values and prevented aggregation at lower concentrations because of their hydrophobicity. The results provide a range of new effective ionic liquid-based additives for higher protein refolding yields and the knowledge of the effect of chemical structures of additives on protein refolding.

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“…Such proteins are e.g. papain, lysozyme, human serum albumin as well as bovine serum albumin (BSA), the object of our studies [16][17][18][19][20][21][22][23]. The majority of the works published in this field deal with the nature of the interaction of these proteins with anionic surfactants, whereas the number of publications addressing the effect of cationic and non-ionic surfactants on various proteins is much lower [24].…”

Section: Introductionmentioning

confidence: 97%

Hydrophobization of bovine serum albumin with cationic surfactants with different hydrophobic chain length

Kun

KIS

Dékány

2010

Colloids and Surfaces B: Biointerfaces

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How CTAB assists the refolding of native and recombinant lysozyme

Cited by 37 publications

References 31 publications

Refolding of denatured/reduced lysozyme at high concentrations by artificial molecular chaperone‐ion exchange chromatography

Refolding of denatured/reduced lysozyme at high concentrations by artificial molecular chaperone‐ion exchange chromatography

Protein Refolding by N-Alkylpyridinium and N-Alkyl-N-methylpyrrolidinium Ionic Liquids

Hydrophobization of bovine serum albumin with cationic surfactants with different hydrophobic chain length

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