Fusion gene consisting of dextran-binding domain from Leuconostoc mesenteroides subsp. Mesenteroides (DBD) and human recombinant interferon-beta (IFN-beta) incorporated between the nucleotide sequence encoding for the recognition site of human enteropeptidase (DDDDK) was installed and constructed in Escherichia coli. The overproducing strain of the chimeric protein DBD-IFN-beta consisting of the IFN-beta, spacer including 10 GS-repeats, human enteropeptidase recognition site, and dextran-binding domain from Leuconostoc mesenteroides was constructed. Free human recombinant interferon-beta was obtained as a result of treatment of the chimeric protein DBD-IFN-beta immobilized on sephadex G-25 with human enteropeptidase. The ability of free and immobilized protein to protect human cells from viral infection was demonstrated. The developed approach can be used for purification of the recombinant proteins with different biological activity and possible construction of new immunostimulating and antiviral drugs, growth factors, anti-cancer drugs, etc.

Download full-text PDF

Source

Publication Analysis

Top Keywords

human enteropeptidase
12
dextran-binding domain
8
domain leuconostoc
8
leuconostoc mesenteroides
8
human recombinant
8
recombinant interferon-beta
8
recognition site
8
chimeric protein
8
protein dbd-ifn-beta
8
human
6

Similar Publications

Coagulation factor XIa (FXIa) is associated with a low risk of bleeding and has been identified as an effective and safe target for the development of novel anticoagulant drugs. In this study, we established an ultrasensitive competitive dual-enzyme cascade signal amplification method for the quantitative analysis and screening of FXIa inhibitors. Due to the specific recognition of FXIa's active site by the aptamer AptE40, the AptE40-QDs-EK recognition probe modified with enterokinase (EK) and the aptamer AptE40, was attached to the MNPs-FXIa capture probe.

View Article and Find Full Text PDF

Metformin inhibits digestive proteases and impairs protein digestion in mice.

J Biol Chem

December 2023

Microbial Sciences Institute, Yale University, West Haven, Connecticut, USA; Department of Microbial Pathogenesis, Yale University School of Medicine, New Haven, Connecticut, USA. Electronic address:

Metformin is among the most prescribed medications worldwide and the first-line therapy for type 2 diabetes. However, gastrointestinal side effects are common and can be dose limiting. The total daily metformin dose frequently reaches several grams, and poor absorption results in high intestinal drug concentrations.

View Article and Find Full Text PDF
Article Synopsis
  • Chemokines are crucial proteins in the immune system that help regulate inflammation by attracting leukocytes; targeting and inhibiting them is a key anti-inflammatory strategy.
  • The study describes a method for producing fluorescently labeled chemokines using recombinant DNA techniques and custom enzymes, which is cost-effective compared to commercial reagents.
  • The resulting fluorescent chemokine (vMIP-fluor) was successfully used in binding studies, showcasing its potential in anti-inflammatory therapies and demonstrating its effectiveness in competition assays with other chemokines.
View Article and Find Full Text PDF

High yield expression in Pichia pastoris of human neutrophil elastase fused to cytochrome B5.

Protein Expr Purif

June 2023

Queensland University of Technology, Molecular Simulation Group, Institute of Health and Biomedical Innovation, Corner Blamey Street & Musk Avenue, Kelvin Grove Urban Village, Queensland, 4059, Australia.

Recombinant human neutrophil elastase (rHNE), a serine protease, was expressed in Pichia pastoris. Glycosylation sites were removed via bioengineering to prevent hyper-glycosylation (a common problem with this system) and the cDNA was codon optimized for translation in Pichia pastoris. The zymogen form of rHNE was secreted as a fusion protein with an N-terminal six histidine tag followed by the heme binding domain of Cytochrome B5 (CytB5) linked to the N-terminus of the rHNE sequence via an enteropeptidase cleavage site.

View Article and Find Full Text PDF
Article Synopsis
  • - Enterokinase deficiency (EKD) is a rare inherited disorder caused by mutations in the TMPRSS15 gene, with only 12 known cases reported so far.
  • - A new case involved a female infant with a specific mutation (c.1216C>T, p.R406*) exhibiting skin lesions similar to acrodermatitis enteropathica (AE).
  • - After starting a protein-rich hydrolyzed formula, the infant's skin condition improved significantly in just 11 days, indicating that AE-like rashes can be linked to EKD and broadening the understanding of TMPRSS15 mutations.
View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!