Comparison of E. coli based self-inducible expression systems containing different human heat shock proteins

Abstract IPTG-inducible promoter is popularly used for the expression of recombinant proteins. However, it is not suitable at the industrial scale due to the high cost and toxicity on the producing cells. Recently, a Self-Inducible Expression (SILEX) system has developed to bypass such problems usin...

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Autores principales:	Fatemeh Sadat Shariati, Malihe Keramati, Vahideh Valizadeh, Reza Ahangari Cohan, Dariush Norouzian
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e299f1fcf6914aabbb490ac40c59350e
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spelling	oai:doaj.org-article:e299f1fcf6914aabbb490ac40c59350e2021-12-02T11:37:18ZComparison of E. coli based self-inducible expression systems containing different human heat shock proteins10.1038/s41598-021-84188-82045-2322https://doaj.org/article/e299f1fcf6914aabbb490ac40c59350e2021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-84188-8https://doaj.org/toc/2045-2322Abstract IPTG-inducible promoter is popularly used for the expression of recombinant proteins. However, it is not suitable at the industrial scale due to the high cost and toxicity on the producing cells. Recently, a Self-Inducible Expression (SILEX) system has developed to bypass such problems using Hsp70 as an autoinducer. Herein, the effect of other heat shock proteins on the autoinduction of green fluorescent protein (EGFP), romiplostim, and interleukin-2 was investigated. For quantitative measurements, EGFP expression was monitored after double-transformation of pET28a-EGFP and pET21a-(Hsp27/Hsp40/Hsp70) plasmids into E. coli using fluorimetry. Moreover, the expression level, bacterial growth curve, and plasmid and expression stability were compared to an IPTG- inducible system using EGFP. Statistical analysis revealed a significant difference in EGFP expression between autoinducible and IPTG-inducible systems. The expression level was higher in Hsp27 system than Hsp70/Hsp40 systems. However, the highest amount of expression was observed for the inducible system. IPTG-inducible and Hsp70 systems showed more lag-time in the bacterial growth curve than Hsp27/Hsp40 systems. A relatively stable EGFP expression was observed in SILEX systems after several freeze–thaw cycles within 90 days, while, IPTG-inducible system showed a decreasing trend compared to the newly transformed bacteria. Moreover, the inducible system showed more variation in the EGFP expression among different clones than clones obtained by SILEX systems. All designed SILEX systems successfully self-induced the expression of protein models. In conclusion, Hsp27 system could be considered as a suitable autoinducible system for protein expression due to less metabolic burden, lower variation in the expression level, suitable plasmid and expression stability, and a higher expression level.Fatemeh Sadat ShariatiMalihe KeramatiVahideh ValizadehReza Ahangari CohanDariush NorouzianNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Fatemeh Sadat Shariati Malihe Keramati Vahideh Valizadeh Reza Ahangari Cohan Dariush Norouzian Comparison of E. coli based self-inducible expression systems containing different human heat shock proteins
description	Abstract IPTG-inducible promoter is popularly used for the expression of recombinant proteins. However, it is not suitable at the industrial scale due to the high cost and toxicity on the producing cells. Recently, a Self-Inducible Expression (SILEX) system has developed to bypass such problems using Hsp70 as an autoinducer. Herein, the effect of other heat shock proteins on the autoinduction of green fluorescent protein (EGFP), romiplostim, and interleukin-2 was investigated. For quantitative measurements, EGFP expression was monitored after double-transformation of pET28a-EGFP and pET21a-(Hsp27/Hsp40/Hsp70) plasmids into E. coli using fluorimetry. Moreover, the expression level, bacterial growth curve, and plasmid and expression stability were compared to an IPTG- inducible system using EGFP. Statistical analysis revealed a significant difference in EGFP expression between autoinducible and IPTG-inducible systems. The expression level was higher in Hsp27 system than Hsp70/Hsp40 systems. However, the highest amount of expression was observed for the inducible system. IPTG-inducible and Hsp70 systems showed more lag-time in the bacterial growth curve than Hsp27/Hsp40 systems. A relatively stable EGFP expression was observed in SILEX systems after several freeze–thaw cycles within 90 days, while, IPTG-inducible system showed a decreasing trend compared to the newly transformed bacteria. Moreover, the inducible system showed more variation in the EGFP expression among different clones than clones obtained by SILEX systems. All designed SILEX systems successfully self-induced the expression of protein models. In conclusion, Hsp27 system could be considered as a suitable autoinducible system for protein expression due to less metabolic burden, lower variation in the expression level, suitable plasmid and expression stability, and a higher expression level.
format	article
author	Fatemeh Sadat Shariati Malihe Keramati Vahideh Valizadeh Reza Ahangari Cohan Dariush Norouzian
author_facet	Fatemeh Sadat Shariati Malihe Keramati Vahideh Valizadeh Reza Ahangari Cohan Dariush Norouzian
author_sort	Fatemeh Sadat Shariati
title	Comparison of E. coli based self-inducible expression systems containing different human heat shock proteins
title_short	Comparison of E. coli based self-inducible expression systems containing different human heat shock proteins
title_full	Comparison of E. coli based self-inducible expression systems containing different human heat shock proteins
title_fullStr	Comparison of E. coli based self-inducible expression systems containing different human heat shock proteins
title_full_unstemmed	Comparison of E. coli based self-inducible expression systems containing different human heat shock proteins
title_sort	comparison of e. coli based self-inducible expression systems containing different human heat shock proteins
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/e299f1fcf6914aabbb490ac40c59350e
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Comparison of E. coli based self-inducible expression systems containing different human heat shock proteins

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