Localization and activity of recombinant streptavidin in cell fractions of Escherichia coli
- Authors: Zotova DV1, Grudinina NA1, Antimonova OI1, Shavlovsky MM2, Polyakov DS1
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Affiliations:
- Institute of Experimental Medicine
- North-Western State Medical University named after I.I. Mechnikov
- Issue: Vol 18, No 3 (2018)
- Pages: 69-76
- Section: Articles
- Published: 15.09.2018
- URL: https://journals.eco-vector.com/MAJ/article/view/10935
- DOI: https://doi.org/10.17816/MAJ18369-76
- ID: 10935
Cite item
Abstract
The authors provide a genetic construct for obtaining recombinant biologically active wild type streptavidin with high yield. Addition of leader peptide and oligohistidine sequence to the streptavidin sequence makes it possible to isolate soluble streptavidin from the culture medium and from the soluble fraction. Temperature conditions for inducing of protein synthesis were found to have a significant effect on the distribution profile of streptavidin between fractions. The obtained protein product is not contaminated with endogenous biotin. The provided method excludes denaturation and renaturation steps which are necessary for isolation of the desired product from inclusion bodies but substantially reduce the yield. Thus, the provided approach allows to increase the yield of biologically highly active strepatividin which is capable of binding biotin and biotin-containing compounds and can be used for various practical purposes.
Full Text
About the authors
D V Zotova
Institute of Experimental Medicine
N A Grudinina
Institute of Experimental Medicine
O I Antimonova
Institute of Experimental Medicine
M M Shavlovsky
North-Western State Medical University named after I.I. Mechnikov
D S Polyakov
Institute of Experimental Medicine
References
- Zhang M, Shao J, Xiao J, et al. A novel approach to make homogeneous protease-stable monovalent streptavidin. Biochem Biophys Res Commun. 2015;463(4):1059-1063. doi: 10.1016/j.bbrc.2015.06.058.
- Green NM. Avidin and streptavidin. Methods Enzymol. 1990;184:51-67. doi: 10.1016/0076-6879(90)84259-j.
- Syrkina MS, Shirokov DA, Rubtsov MA, et al. Preparation and functional evaluation of RGD-modified streptavidin targeting to integrin-expressing melanoma cells. Protein Eng Des Sel. 2013;26(2):143-150. doi: 10.1093/protein/gzs076.
- Hofmann K, Wood SW, Brinton CC, et al. Iminobiotin affinity columns and their application to retrieval of streptavidin. Proc Natl Acad Sci U S A. 1980;77(8):4666-4668. doi: 10.1073/pnas.77.8.4666.
- Поляков Д.С., Грудинина Н.А., Соловьев К.В., и др. Бета2-микроглобулиновый амилоидоз: фибриллогенез природного и рекомбинантных бета2-микроглобулинов человека // Медицинский академический журнал. - 2010. - Т. 10. - № 2. - С. 40-49. [Polyakov DS, Grudinina NA, Solov’ev KV, et al. Bela2-microglobuline amyloidosis: Fibrillogenesis of natural and recombinant human beta2-microglobulines. Medical academic journal. 2010;10(2);40-49 (In Russ.)]
- Ausubel FM, Brent R, Kingston RE, et al. Current Protocols in Molecular Biology. New York: John Wiley & Sons; 1989.
- Поляков Д.С., Грудинина Н.А., Соловьев К.В., и др. Получение рекомбинантного β2-микроглобулина человека и его фибриллогенез при низких и нейтральных значениях рН // Молекулярная медицина. - 2011. - № 2. - C. 36-39. [Polyakov DS, Grudinina NA, Solov’ev KV, et al. Production of recombinant human β2-microglobulin and its amyloid fibril formation at acidic and neutral pH. Molekuliarnaia meditsina. 2011;(2);36-39. (In Russ.)]
- Solovyov KV, Polyakov DS, Grudinina NA, et al. Expression in E. coli and purification of the fibrillogenic fusion proteins TTR-sfGFP and beta2M-sfGFP. Prep Biochem Biotechnol. 2011;41(4):337-349. doi: 10.1080/10826068.2010.548433.
- Поляков Д.С., Антимонова О.И., Сахабеев Р.Г., и др. Влияние наночастиц из полимолочной кислоты на иммуногенность связанного с ним белка // Инфекция и иммунитет. - 2017. - Т. 7. - № 2. - С. 123-129. [Polyakov DS, Antimonova OI, Sakhabeev RG, et al. Polylactic acid nanoparticles influence on immunogenicity of the protein bound with them. Russian Journal of Infection and Immunity. 2017;7(2):123-129. (In Russ.)] doi: 10.15789/2220-7619-2017-2-123-129.
- Solovyov KV, Kern АM, Grudinina NA, et al. Genetic structures and conditions of their expression, which allow receiving native recombinant proteins with high output. International Journal of biomedicine. 2012;2(1):45-49.
- Антимонова О.И., Грудинина Н.А., Егоров В.В., и др. Взаимодействие красителя конго красный с фибриллами лизоцима, бета2-микроглобулина и транстиретина // Цитология. - 2016. - Т. 58. - № 2. - С. 156-163. [Antimonova OI, Grudinina NA, Egorov VV, et al. Interaction of the dye congo red with fibrils of lysozyme, beta2-microglobultn and transthyretin. Cell and tissue biology. 2016;58(2):156-163. (In Russ.)]
- Поляков Д.С., Сахабеев Р.Г., Шавловский М.М. Частичная денатурация рекомбинантного белка для его аффинного выделения // Прикладная биохимия и микробиология. - 2016. - Т. 52. - № 1. - С. 122-127. [Polyakov DS, Sakhabeev RG, Shavlovskiy MM. Polylactic acid nanoparticles influence on immunogenicity of the protein bound with them. Applied biochemistry and microbiology. 2016;52(1):122-127. (In Russ.)]. doi: 10.7868/S0555109916010104.