Engineering Lactobacillus plantarum Services for Live Biotherapeutics Drug Discovery
Background Services Advantages FAQs Published Data Resources
Background
Due to the widespread application of lactic acid bacteria (LAB) in industrial and health fields, a large amount of research is devoted to the development and modification of LAB-induced exogenous protein expression systems to achieve various goals, including the production and secretion of eukaryotic immune proteins. Lactobacillus provides a relatively safe carrier for microbial treatment of various health problems. Lactobacillus plantarum (L. plantarum) and many other LABs are generally considered safe (GRAS) microorganisms with the ability to efficiently secrete recombinant proteins directly in the culture medium. Therefore, in the fields of medicine and industrial biotechnology, they are considered emerging candidates for recombinant protein expression and genetic and metabolic cell engineering. Scientists have described some tools that have created interesting potential for using L. plantarum engineering as a synthetic biology solution for industrial and medical environments. The construction and induction of gene expression systems through promoter screening, adaptation of TCS, and attempts at repressor engineering have been the greatest advances in L. plantarum.
Fig.1 Scheme showing the inducible protein expression systems in L. plantarum.1
L. plantarum Genetic Engineering Services
Services
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Gene knock-out
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Gene knock-in/Gene insertion
Deliverables
Engineered strain in glycerol stock
Quality Control
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Sequencing Validation
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Western Blot Validation (Optional)
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PAGE
or cell-based assay of your interest.
Delivery Time
Starting from 4-6 weeks.
Materials from Clients
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Overall construction scheme.
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Host strain
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Target gene name
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Target gene sequence, if the whole genome sequence is unavailable
How L. plantarum Strain Construction Service Can Assist Your Project
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Construct the target strains according to the needs of clients or refer to the published scheme.
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Optimize the program to improve the success rate.
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Delivery of the constructed target strains (knockout or integrated).
Advantages
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L. plantarum strains have been qualified with a GRAS status by the USFDA.
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They are facultatively anaerobic or microaerophilic and are easy to culture.
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They are effective probiotics to alleviate disease.
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The genomes of a variety of strains have been sequenced, leading to a better understanding of their properties and functions.
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They are genetically susceptible, and several genetic components, including promoters, plasmid replicon, plasmid retention system, secreted signal peptides, and gene-editing tools, have been shown to work reliably and stably.
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Heterologous expression and secretion ability of recombinant proteins from L. plantarum strains.
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The expression characteristics of inducible genes and their protein secretion ability prompted the exploration of heterologous expression of recombinant proteins in L. plantarum strains.
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Engineering Lactobacilli exosomes for enhanced functionality.
Diversity of Gene Cassettes of the L. plantarum Strains
In general, the gene order is very conserved. However, some examples of highly variable regions in the chromosome are given below.
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Highly Variable Cassettes
Prophages, IS elements and transposases.
Plantaricin biosynthesis genes.
CPS/EPS biosynthesis genes.
A large number of gene cassettes in L. plantarum appear to be directly connected, so further understanding of the molecular mechanisms of acquisition and assembly of these gene cassettes encoding functional pathways in L. plantarum is important for better utilization of this natural metabolic engineering.
Frequently Asked Questions
Can L. plantarum be developed as a platform for mucosal vaccines?
The gut microbiota plays a crucial role in antiviral immunity, and among these beneficial microbes with immunomodulatory capabilities, some L. plantarum strains are interesting alternatives to improve local and distal antiviral immune responses during oral administration. Appropriate L. plantarum strains with immune activity can simultaneously enhance the antiviral immunity of both intestinal and respiratory mucosa, making them an interesting biotechnology resource for developing mucosal vaccines.
Published Data
Genetically Engineered L. plantarum WCFS1 Constitutively Secreting Heterologous Oxalate Decarboxylase and Degrading Oxalate Under In Vitro
Abstract
Hyperoxaluria is a major risk factor for urinary stone diseases, and calcium oxalate (CaOx) is the most common type of kidney stone. The recombinant L. plantarum containing pLdhl0373OxdC could constitutively secrete bioactive OxdC and degrade oxalic acid in vitro.3
The food-grade expression system of L. plantarum using β-galactosidase small subunit as selection marker and lactose as screening condition
Abstract
The construction and application of L. plantarum food-grade expression system has gradually shown significant advantages in food and medicine fields. This study successfully established a food-grade expression system of L. plantarum with lacM gene as the selective marker and lactose as the screening condition, which laid a foundation for the application of L. plantarum.
Fig.2 Schematic diagram of the construction of food-grade expression vector pMG36m.2
Resources
The development of genetically modified and biologically contained OxdC-secreting L. plantarum with improved GI survival will enable intestinal oxalate degradation beneficial in humans for mitigation of hyperoxaluria, which provides a potential therapeutic importance for degrading intestinal oxalate. Creative Biolabs has accumulated extensive experience in the development of live biotherapeutics and is familiar with a variety of probiotics. If you are interested in our L. plantarum engineering strains construction services, please do not hesitate to contact us for more details, we look forward to helping you at some stage of the project.
References
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Blanch-Asensio, Marc, et al. "Expanding the genetic programmability of Lactiplantibacillus plantarum." Microbial Biotechnology 17.1 (2024): e14335.
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Wang, Ting, et al. "Food-grade expression system of Lactobacillus plantarum using β-galactosidase small subunit as selection marker and lactose as screening condition." LWT 182 (2023): 114922.
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Sasikumar, Ponnusamy, et al. "Genetically engineered Lactobacillus plantarum WCFS1 constitutively secreting heterologous oxalate decarboxylase and degrading oxalate under in vitro." Current microbiology 69 (2014): 708-715.
For Research Use Only. Not intended for use in food manufacturing or medical procedures (diagnostics or therapeutics). Do Not Use in Humans.
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