Solubilization and Refolding of Inclusion Body Proteins.
Combinatorial mild solubilization method
Inclusion bodies
Protein aggregates
Protein refolding
Solubilization
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
entrez:
28
1
2022
pubmed:
29
1
2022
medline:
31
3
2022
Statut:
ppublish
Résumé
Expression of heterologous proteins in E. coli often leads to the formation of protein aggregates known as inclusion bodies (IBs). Inclusion body aggregates pose a major hurdle in the recovery of bioactive proteins from E. coli. Usage of strong denaturing buffers for solubilization of bacterial IBs results in poor recovery of bioactive protein. Structure-function understanding of IBs in the last two decades have led to the development of several mild solubilization buffers, which improve the recovery of bioactive from IBs. Recently, combinatorial mild solubilization methods have paved the way for solubilization of wide range of inclusion bodies with appreciable refolding yield. Here, we describe a simple protocol for solubilization and refolding of an inclusion body protein with appreciable recovery.
Identifiants
pubmed: 35089569
doi: 10.1007/978-1-0716-1859-2_22
doi:
Substances chimiques
Proteins
0
Recombinant Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
371-387Informations de copyright
© 2022. Springer Science+Business Media, LLC, part of Springer Nature.
Références
Slouka C, Kopp J, Hutwimmer S et al (2018) Custom made inclusion bodies: impact of classical process parameters and physiological parameters on inclusion body quality attributes. Microb Cell Factories 17:148
Jürgen B, Breitenstein A, Urlacher V et al (2010) Quality control of inclusion bodies in Escherichia coli. Microb Cell Factories 9:41
Singh A, Upadhyay V, Upadhyay AK et al (2015) Protein recovery from inclusion bodies of Escherichia coli using mild solubilization process. Microb Cell Factories 14:41
Singh SM, Sharma A, Upadhyay AK et al (2012) Solubilization of inclusion body proteins using n-propanol and its refolding into bioactive form. Protein Expr Purif 81:75–82
pubmed: 21964443
Dill KA, Shortle D (1991) Denatured states of proteins. Annu Rev Biochem 60:795–825
pubmed: 1883209
Rudolph R, Lilie H (1996) In vitro folding of inclusion body proteins. FASEB J 10:49–56
pubmed: 8566547
Anfinsen CB (1973) Principles that govern the folding of protein chains. Science 181:223–230
pubmed: 4124164
Singhvi P, Saneja A, Srichandan S et al (2020) Bacterial inclusion bodies: a treasure trove of bioactive proteins. Trends Biotechnol 38:474–486
pubmed: 31954528
Chung WJ, Huang CL, Gong HY et al (2015) Recombinant production of biologically active giant grouper (Epinephelus lanceolatus) growth hormone from inclusion bodies of Escherichia coli by fed-batch culture. Protein Expr Purif 110:79–88
pubmed: 25703054
Malavasi NV, Foguel D, Bonafe CFS et al (2011) Protein refolding at high pressure: optimization using eGFP as a model. Process Biochem 46:512–518
Berini F, Presti I, Beltrametti F et al (2017) Production and characterization of a novel antifungal chitinase identified by functional screening of a suppressive-soil metagenome. Microb Cell Factories 16:16
Mohammadian A, Kaghazian H, Kavianpour A et al (2018) Solubilization of inclusion body proteins using low and very low concentrations of chemicals: implications of novel combined chemical treatment designs in enhancement of post-solubilization target protein purity and biological activity. J Chem Technol Biotechnol 93:1579–1587
Panda AK, Seikh GB Eshwari ANS et al (2007) A process for solubilization of recombinant protein expressed as inclusion body. US patent number US7189811B2
Upadhyay V, Singh A, Jha D et al (2016) Recovery of bioactive protein from bacterial inclusion bodies using trifluoroethanol as solubilization agent. Microb Cell Factories 15:100
Park AR, Jang SW, Kim JS et al (2018) Efficient recovery of recombinant CRM197 expressed as inclusion bodies in E. coli. PLoS One 13:e0201060
pubmed: 30021008
pmcid: 6051658
Qi X, Sun Y, Xiong S (2015) A single freeze-thawing cycle for highly efficient solubilization of inclusion body proteins and its refolding into bioactive form. Microb Cell Factories 14:24
Chura-Chambi RM, Rosa Da Silva CM, Pereira LR et al (2019) Protein refolding based on high hydrostatic pressure and alkaline pH: application on a recombinant dengue virus NS1 protein. PLoS One 14:e0211162
pubmed: 30682103
pmcid: 6347194
Padhiar AA, Chanda W, Joseph TP et al (2018) Comparative study to develop a single method for retrieving wide class of recombinant proteins from classical inclusion bodies. Appl Microbiol Biotechnol 102:2363–2377
pubmed: 29387954
Wang Q, Liu Y, Zhang C et al (2017) High hydrostatic pressure enables almost 100% refolding of recombinant human ciliary neurotrophic factor from inclusion bodies at high concentration. Protein Expr Purif 133:152–159
pubmed: 28323167
Mirzadeh A, Saadatnia G, Golkar M et al (2017) Production of refolded Toxoplasma gondii recombinant SAG1-related sequence 3 (SRS3) and its use for serodiagnosis of human toxoplasmosis. Protein Expr Purif 133:66–74
pubmed: 28263855
Pan S, Odabas N, Sissolak B et al (2015) Engineering batch and pulse refolding with transition of aggregation kinetics: an investigation using green fluorescent protein (GFP). Chem Eng Sci 131:91–100
García-Fruitós E, Vázquez E, Díez-Gil C et al (2012) Bacterial inclusion bodies: making gold from waste. Trends Biotechnol 30:65–70
pubmed: 22037492
Ramón A, Señorale-Pose M, Marín M (2014) Inclusion bodies: not that bad. Front Microbiol 5:56
pubmed: 24592259
pmcid: 3924032
Jevševar S, Gaberc-Porekar V, Fonda I et al (2005) Production of nonclassical inclusion bodies from which correctly folded protein can be extracted. Biotechnol Prog 21:632–639
pubmed: 15801811
Pesarrodona M, Fernández Y, Foradada L et al (2016) Conformational and functional variants of CD44-targeted protein nanoparticles bio-produced in bacteria. Biofabrication 8:025001
pubmed: 27078873
Kumar L, Colomb W, Czerski J et al (2018) Efficient protease based purification of recombinant matrix metalloprotease-1 in E. coli. Protein Expr Purif 148:59–67
pubmed: 29626520
Stark GR, Stein WH, Moore S (1960) Reactions of the cyanate present in aqueous urea with amino acids and proteins. J Biol Chem 235:3177–3181
Yang Z, Zhang L, Zhang Y et al (2011) Highly efficient production of soluble proteins from insoluble inclusion bodies by a two-step-denaturing and refolding method. PLoS One 6:e22981
pubmed: 21829569
pmcid: 3146519
Feng Y, Liu L, Wang J et al (2012) Integrated refolding techniques for Schistosoma japonicum MTH1 overexpressed as inclusion bodies in Escherichia coli. Protein Expr Purif 84:181–187
pubmed: 22641057
Gifre-Renom L, Cano-Garrido O, Fàbregas F et al (2018) A new approach to obtain pure and active proteins from Lactococcus lactis protein aggregates. Sci Rep 8:13917
pubmed: 30224788
pmcid: 6141594
Hagel P, Gerding JJT, Fieggen W et al (1971) Cyanate formation in solutions of urea. I. Calculation of cyanate concentrations at different temperature and pH. Biochim Biophys Acta 243:366–373
pubmed: 5129584
Noji M, So M, Yamaguchi K et al (2018) Heat-induced aggregation of hen ovalbumin suggests a key factor responsible for serpin polymerization. Biochemistry 57:5415–5426
pubmed: 30148614
Alam MT, Rizvi A, Rauf MA et al (2018) Thermal unfolding of human lysozyme induces aggregation: recognition of the aggregates by antisera against the native protein. Int J Biol Macromol 113:976–982
pubmed: 29462680
Datta I, Gautam S, Gupta MN (2013) Microwave assisted solubilization of inclusion bodies. Sustain Chem Process 1:2
Singh A, Upadhyay V, Panda AK (2013) N-propanol based solubilization buffer enhances refolding yield of inclusion body protein by populating intermediates to the folding pathway. Biophys J 104:396a
Yu Y, Wang J, Shao Q et al (2016) The effects of organic solvents on the folding pathway and associated thermodynamics of proteins: a microscopic view. Sci Rep 6:19500
pubmed: 26775871
pmcid: 4726029
Perham M, Liao J, Wittung-Stafshede P (2006) Differential effects of alcohols on conformational switchovers in α-helical and β-sheet protein models. Biochemistry 45:7740–7749
pubmed: 16784225
Li A, Sowder RC, Henderson LE et al (2001) Chemical cleavage at aspartyl residues for protein identification. Anal Chem 73:5395–5402
pubmed: 11816565
Hauptmann A, Podgoršek K, Kuzman D et al (2018) Impact of buffer, protein concentration and sucrose addition on the aggregation and particle formation during freezing and thawing. Pharm Res 35:101
pubmed: 29556730
pmcid: 5859698
Quaas B, Burmeister L, Li Z et al (2018) Properties of dimeric, disulfide-linked rhBMP-2 recovered from E. coli derived inclusion bodies by mild extraction or chaotropic solubilization and subsequent refolding. Process Biochem 67:80–87
Upadhyay AK, Singh A, Mukherjee KJ et al (2014) Refolding and purification of recombinant L-asparaginase from inclusion bodies of E. coli into active tetrameric protein. Front Microbiol 5:486
pubmed: 25309524
pmcid: 4164012
Singh SM, Panda AK (2005) Solubilization and refolding of bacterial inclusion body proteins. J Biosci Bioeng 99:303–310
pubmed: 16233795
Upadhyay AK, Murmu A, Singh A et al (2012) Kinetics of inclusion body formation and its correlation with the characteristics of protein aggregates in Escherichia coli. PLoS One 7:e33951
pubmed: 22479486
pmcid: 3315509
Alibolandi M, Mirzahoseini H (2011) Chemical assistance in refolding of bacterial inclusion bodies. Biochem Res Int 2011:631607
pubmed: 21822494
pmcid: 3148444
Burgess RR (2009) Chapter 17 refolding solubilized inclusion body proteins. In: Methods of enzymology. Academic, pp 259–282