ArticleMicrobial cell factories2007
Efficient production and secretion of bovine beta-lactoglobulin by Lactobacillus casei.
Article in Microbial cell factories, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 55 citations in OpenAlex.
- Exploration of endogenous signal peptides and tandem linkage strategies for enhanced heterologous protein secretion inSynthetic and systems biotechnology · 2026Article
- 2-O-α-D-glucosyl glycerol production by whole-cell biocatalyst of lactobacilli encapsulating sucrose phosphorylase with improved glycerol affinity and conversion rate.Microbial cell factories · 2024Article
- Genome-wide high-throughput signal peptide screening via plasmid pUC256E improves protease secretion in Lactiplantibacillus plantarum and Pediococcus acidilactici.BMC genomics · 2022Article
- K-Ras Peptide Mimotope Induces Antigen Specific Th1 and B-Cell Immune Responses against G12A-Mutated K-Ras Antigen in Balb/c Mice.Vaccines · 2021Article
- Targeted and Repetitive Chromosomal Integration Enables High-Level Heterologous Gene Expression inApplied and environmental microbiology · 2019Article
- Coupling the recombineering to Cre-lox system enables simplified large-scale genome deletion in Lactobacillus casei.Microbial cell factories · 2018Article
- Gut remediation: a potential approach to reducing chromium accumulation using Lactobacillus plantarum TW1-1.Scientific reports · 2017Article
- Development of a counterselectable seamless mutagenesis system in lactic acid bacteria.Microbial cell factories · 2017Article
- Engineered β-Lactoglobulin Produced in E. coli: Purification, Biophysical and Structural Characterisation.Molecular biotechnology · 2016Article
- Secretion of M2e:HBc fusion protein by Lactobacillus casei using Cwh signal peptide.FEMS microbiology letters · 2016Article
- Reduction of Phytate in Soy Drink by Fermentation with Lactobacillus casei Expressing Phytases From Bifidobacteria.Plant foods for human nutrition (Dordrecht, Netherlands) · 2015Article
- Engineered commensal bacteria reprogram intestinal cells into glucose-responsive insulin-secreting cells for the treatment of diabetes.Diabetes · 2015Article
- Construction and application of a food-grade expression system for Lactococcus lactis.Molecular biotechnology · 2013Article
- Heterologous protein display on the cell surface of lactic acid bacteria mediated by the s-layer protein.Microbial cell factories · 2011Article
- Characterization of a cryptic plasmid pD403 from Lactobacillus plantarum and construction of shuttle vectors based on its replicon.Molecular biotechnology · 2010Article
- Characterization of a novel LysM domain from Lactobacillus fermentum bacteriophage endolysin and its use as an anchor to display heterologous proteins on the surfaces of lactic acid bacteria.Applied and environmental microbiology · 2010Article
- High yield recombinant thermostable alpha-amylase production using an improved Bacillus licheniformis system.Microbial cell factories · 2009Article
- Secretion of insulinotropic proteins by commensal bacteria: rewiring the gut to treat diabetes.Applied and environmental microbiology · 2008Article
- In vivo transfer of plasmid from food-grade transiting lactococci to murine epithelial cells.Gene therapy · 2008Article
- Production of Recombinant Peanut Allergen Ara h 2 using Lactococcus lactis.Microbial cell factories · 2007Article
Corrections and comments
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Authors and funding
6 authors at 5 institutions in 3 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundLactic acid bacteria (LAB) are attractive tools to deliver therapeutic molecules at the mucosal level. The model LAB Lactococcus lactis has been intensively used to produce and deliver such heterologous proteins. However, compared to recombinant lactococci, lactobacilli offer some advantages such as better survival in the digestive tract and immunomodulatory properties. Here, we compared different strategies to optimize the production of bovine beta-lactoglobulin (BLG), a major cow's milk allergen, in the probiotic strain Lactobacillus casei BL23.
resultsUsing a nisin-inducible plasmid system, we first showed that L. casei BL23 strain could efficiently secrete a reporter protein, the staphylococcal nuclease (Nuc), with the lactococcal signal peptide SPUsp45 fused to its N-terminus. The fusion of SPUsp45 failed to drive BLG secretion but led to a 10-fold increase of intracellular BLG production. Secretion was significantly improved when the synthetic propeptide LEISSTCDA (hereafter called LEISS) was added to the N-terminus of the mature moiety of BLG. Secretion rate of LEISS-BLG was 6-fold higher than that of BLG alone while intracellular production reached then about 1 mg/L of culture. The highest yield of secretion was obtained by using Nuc as carrier protein. Insertion of Nuc between LEISS and BLG resulted in a 20-fold increase in BLG secretion, up to 27 microg/L of culture. Furthermore, the lactococcal nisRK regulatory genes were integrated into the BL23 chromosome. The nisRK insertion allowed a decrease of BLG synthesis in uninduced cultures while BLG production increased by 50% after nisin induction. Moreover, modification of the induction protocol led to increase the proportion of soluble BLG to around 74% of the total BLG production.
conclusionBLG production and secretion in L. casei were significantly improved by fusions to a propeptide enhancer and a carrier protein. The resulting recombinant strains will be further tested for their ability to modulate the immune response against BLG via mucosal delivery in a cow's milk allergy model in mice.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.