ArticleApplied and environmental microbiology2009
Production of functionalized biopolyester granules by recombinant Lactococcus lactis.
Article in Applied and environmental microbiology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 48 citations in OpenAlex.
- Transcriptional control of carbohydrate catabolism by the CcpA protein in the ruminal bacteriumApplied and environmental microbiology · 2023Article
- Electrochemical Detection of Global DNA Methylation Using Biologically Assembled Polymer Beads.Cancers · 2021Article
- Engineering Bacillus megaterium for production of functional intracellular materials.Microbial cell factories · 2017Article
- Engineering Mycobacteria for the Production of Self-Assembling Biopolyesters Displaying Mycobacterial Antigens for Use as a Tuberculosis Vaccine.Applied and environmental microbiology · 2017Article
- Bioengineering a bacterial pathogen to assemble its own particulate vaccine capable of inducing cellular immunity.Scientific reports · 2017Article
- Smart polyhydroxyalkanoate nanobeads by protein based functionalization.Nanomedicine : nanotechnology, biology, and medicine · 2015Review
- Polyhydroyxalkanoate synthase fusions as a strategy for oriented enzyme immobilisation.Molecules (Basel, Switzerland) · 2014Review
- Relevant uses of surface proteins--display on self-organized biological structures.Microbial biotechnology · 2012Review
- Vaccines displaying mycobacterial proteins on biopolyester beads stimulate cellular immunity and induce protection against tuberculosis.Clinical and vaccine immunology : CVI · 2012Article
- Production of a particulate hepatitis C vaccine candidate by an engineered Lactococcus lactis strain.Applied and environmental microbiology · 2011Article
- In vivo enzyme immobilization by inclusion body display.Applied and environmental microbiology · 2010Article
- Immune responses elicited in mice with recombinant Lactococcus lactis expressing F4 fimbrial adhesin FaeG by oral immunization.Veterinary research communications · 2010Article
- Cell surface of Lactococcus lactis is covered by a protective polysaccharide pellicle.The Journal of biological chemistry · 2010Article
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Authors and funding
3 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Many bacteria are naturally capable of accumulating biopolyesters composed of 3-hydroxy fatty acids as intracellular inclusions, which serve as storage granules. Recently, these inclusions have been considered as nano-/microbeads with surface-attached proteins, which can be engineered to display various protein-based functions that are suitable for biotechnological and biomedical applications. In this study, the food-grade, generally-regarded-as-safe gram-positive organism Lactococcus lactis was engineered to recombinantly produce the biopolyester poly(3-hydroxybutyrate) and the respective intracellular inclusions. The codon-optimized polyhydroxybutyrate biosynthesis operon phaCAB from Cupriavidus necator was expressed using the nisin-controlled gene expression system. Recombinant L. lactis accumulated up to 6% (wt/wt) poly(3-hydroxybutyrate) of cellular dry weight. Poly(3-hydroxybutyrate) granules were isolated and analyzed with respect to bound proteins using biochemical methods and with respect to shape/size using transmission electron microscopy. The immunoglobulin G (IgG) binding ZZ domain of Staphylococcus aureus protein A was chosen as an exemplary functionality to be displayed at the granule surface by fusing it to the N terminus of the granule-associated poly(3-hydroxybutyrate) synthase. The presence of the fusion protein at the surface of isolated granules was confirmed by peptide fingerprinting using matrix-assisted laser desorption ionization-time of flight (mass spectrometry). The functionality of the ZZ domain-displaying granules was demonstrated by enzyme-linked immunosorbent assay and IgG affinity purification. In both assays, the ZZ beads from recombinant L. lactis performed at least equally to ZZ beads from Escherichia coli. Overall, in this study it was shown that recombinant L. lactis can be used to manufacture endotoxin-free poly(3-hydroxybutyrate) beads with surface functionalities that are suitable for biomedical applications.
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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.