NDLI: A novel strategy for protein production using non-classical secretion pathway in Bacillus subtilis

Content Provider	Springer Nature : BioMed Central
Author	Chen, Jingqi Zhao, Liuqun Fu, Gang Zhou, Wenjuan Sun, Yuanxia Zheng, Ping Sun, Jibin Zhang, Dawei
Abstract	Background The Gram-positive bacterium Bacillus subtilis has been widely used as a cell factory for the production of proteins due to its generally regarded as safe (GRAS) nature and secretion capability. Of the known secretory pathways in B. subtilis, the majority of proteins are exported from the cytoplasm by Sec pathway, Tat pathway and ABC transporters, etc. However, the production of heterologous proteins by B. subtilis is unfortunately not that straight forward because of the bottlenecks in classical secretion pathways. The aim of this work is to explore a new method for protein production based on non-classical secretion pathway. Results One d-psicose 3-epimerase (RDPE) which converts d-fructose into d-psicose from Ruminococcus sp. 5_1_39BFAA was successfully and substantially secreted into the extracellular milieu without the direction of signal peptide. Subsequently, we demonstrated that RDPE contained no native signal peptide, and the secretion of RDPE was not dependent on Sec or Tat pathway or due to cell lysis, which indicated that RDPE is a non-classically secreted protein. Then, we attempted to evaluate the possibility of using RDPE as a signal to export eighteen reporter proteins into the culture medium. Five of eleven homologous proteins, two of five heterologous proteins from other bacterium and two heterologous proteins of eukaryotic source were successfully secreted into the extracellular milieu at different secretion levels when they were fused to RDPE mediated by a flexible 21-bp linker to keep a distance between two single proteins. Furthermore, the secretion rates of two fusion proteins (RDPE-DnaK and RDPE-RFP) reached more than 50 %. In addition, most of the fusion proteins retained enzyme or biological activity of their corresponding target proteins, and all of the fusions still had the activity of RDPE. Conclusions We found and identified a heterologous non-classically secreted protein RDPE, and showed that RDPE could direct proteins of various types into the culture medium, and thus non-classical protein secretion pathway can be used as a novel secretion pathway for recombinant proteins. This novel strategy for recombinant protein production is helpful to make B. subtilis as a more ideal cell factory for protein production.
Related Links	https://microbialcellfactories.biomedcentral.com/counter/pdf/10.1186/s12934-016-0469-8.pdf
Ending Page	16
Page Count	16
Starting Page	1
File Format	HTM / HTML
ISSN	14752859
DOI	10.1186/s12934-016-0469-8
Journal	Microbial Cell Factories
Issue Number	1
Volume Number	15
Language	English
Publisher	BioMed Central
Publisher Date	2016-04-28
Access Restriction	Open
Subject Keyword	Applied Microbiology Biotechnology Microbiology Microbial Genetics and Genomics Enzymology Genetic Engineering Bacillus subtilis d-Psicose 3-epimerase Fusions Non-classical secretion pathway Localization
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

Highly efficient production of Clostridium cellulolyticum H10 d-psicose 3-epimerase in Bacillus subtilis and use of these cells to produce d-psicose

A new potential secretion pathway for recombinant proteins in Bacillus subtilis

Expression of d-psicose-3-epimerase from Clostridium bolteae and Dorea sp. and whole-cell production of d-psicose in Bacillus subtilis

Production, purification, characterization, and safety evaluation of constructed recombinant D-psicose 3-epimerase

High-level intra- and extra-cellular production of d-psicose 3-epimerase via a modified xylose-inducible expression system in Bacillus subtilis

Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis

Contribution of single amino acid and codon substitutions to the production and secretion of a lipase by Bacillus subtilis

Efficient D-allulose synthesis under acidic conditions by auto-inducing expression of the tandem D-allulose 3-epimerase genes in Bacillus subtilis

A novel strategy for protein production using non-classical secretion pathway in Bacillus subtilis

A novel strategy for protein production using non-classical secretion pathway in Bacillus subtilis

Similar Documents

Highly efficient production of Clostridium cellulolyticum H10 d-psicose 3-epimerase in Bacillus subtilis and use of these cells to produce d-psicose

A new potential secretion pathway for recombinant proteins in Bacillus subtilis

Expression of d-psicose-3-epimerase from Clostridium bolteae and Dorea sp. and whole-cell production of d-psicose in Bacillus subtilis

Production, purification, characterization, and safety evaluation of constructed recombinant D-psicose 3-epimerase

High-level intra- and extra-cellular production of d-psicose 3-epimerase via a modified xylose-inducible expression system in Bacillus subtilis

Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis

Contribution of single amino acid and codon substitutions to the production and secretion of a lipase by Bacillus subtilis

Efficient D-allulose synthesis under acidic conditions by auto-inducing expression of the tandem D-allulose 3-epimerase genes in Bacillus subtilis

A novel strategy for protein production using non-classical secretion pathway in Bacillus subtilis

A novel strategy for protein production using non-classical secretion pathway in Bacillus subtilis