NDLI: Multimodal approaches for the improvement of the cellular folding of a recombinant iron regulatory protein in E. coli

Content Provider	Springer Nature : BioMed Central
Author	Ravitchandirane, Gayathri Bandhu, Sheetal Chaudhuri, Tapan K.
Abstract	Background During the recombinant protein expression, most heterologous proteins expressed in E. coli cell factories are generated as insoluble and inactive aggregates, which prohibit E. coli from being employed as an expression host despite its numerous advantages and ease of use. The yeast mitochondrial aconitase protein, which has a tendency to aggregate when expressed in E. coli cells in the absence of heterologous chaperones GroEL/ES was utilised as a model to investigate how the modulation of physiological stimuli in the host cell can increase protein solubility. The presence of folding modulators such as exogenous molecular chaperones or osmolytes, as well as process variables such as incubation temperature, inducer concentrations, growth media are all important for cellular folding and are investigated in this study. This study also investigated how the cell's stress response system activates and protects the proteins from aggregation. Results The cells exposed to osmolytes plus a pre-induction heat shock showed a substantial increase in recombinant aconitase activity when combined with modulation of process conditions. The concomitant GroEL/ES expression further assists the folding of these soluble aggregates and increases the functional protein molecules in the cytoplasm of the recombinant E. coli cells. Conclusions The recombinant E. coli cells enduring physiological stress provide a cytosolic environment for the enhancement in the solubility and activity of the recombinant proteins. GroEL/ES-expressing cells not only aided in the folding of recombinant proteins, but also had an effect on the physiology of the expression host. The improvement in the specific growth rate and aconitase production during chaperone GroEL/ES co-expression is attributed to the reduction in overall cellular stress caused by the expression host's aggregation-prone recombinant protein expression. Graphical Abstract
Related Links	https://microbialcellfactories.biomedcentral.com/counter/pdf/10.1186/s12934-022-01749-w.pdf
Ending Page	19
Page Count	19
Starting Page	1
File Format	HTM / HTML
ISSN	14752859
DOI	10.1186/s12934-022-01749-w
Journal	Microbial Cell Factories
Issue Number	1
Volume Number	21
Language	English
Publisher	BioMed Central
Publisher Date	2022-02-05
Access Restriction	Open
Subject Keyword	Applied Microbiology Biotechnology Microbiology Microbial Genetics and Genomics Enzymology Genetic Engineering Cellular protein folding Protein aggregation Yeast mitochondrial aconitase Molecular chaperone Cellular stress GroEL/ES Inclusion bodies
Content Type	Text
Resource Type	Article
Subject	Applied Microbiology and Biotechnology Bioengineering Biotechnology
Journal Impact Factor	4.3/2023
5-Year Journal Impact Factor	5.5/2023

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Nucleic acids in inclusion bodies obtained from E. coli cells expressing human interferon-gamma

Yeast prions form infectious amyloid inclusion bodies in bacteria

Using bacterial inclusion bodies to screen for amyloid aggregation inhibitors

Inclusion bodies as potential vehicles for recombinant protein delivery into epithelial cells

Application of an E. coli signal sequence as a versatile inclusion body tag

A variant of green fluorescent protein exclusively deposited to active intracellular inclusion bodies

Mammalian prion protein (PrP) forms conformationally different amyloid intracellular aggregates in bacteria

Influence of pH control in the formation of inclusion bodies during production of recombinant sphingomyelinase-D in Escherichia coli

Kinetics of aggregation and structural properties of proteins in inclusion bodies studied by Fourier transform infrared spectroscopy

Multimodal approaches for the improvement of the cellular folding of a recombinant iron regulatory protein in E. coli

Similar Documents

Nucleic acids in inclusion bodies obtained from E. coli cells expressing human interferon-gamma

Yeast prions form infectious amyloid inclusion bodies in bacteria

Using bacterial inclusion bodies to screen for amyloid aggregation inhibitors

Inclusion bodies as potential vehicles for recombinant protein delivery into epithelial cells

Application of an E. coli signal sequence as a versatile inclusion body tag

A variant of green fluorescent protein exclusively deposited to active intracellular inclusion bodies

Mammalian prion protein (PrP) forms conformationally different amyloid intracellular aggregates in bacteria

Influence of pH control in the formation of inclusion bodies during production of recombinant sphingomyelinase-D in Escherichia coli

Kinetics of aggregation and structural properties of proteins in inclusion bodies studied by Fourier transform infrared spectroscopy

Multimodal approaches for the improvement of the cellular folding of a recombinant iron regulatory protein in E. coli