Bile-induced promoters for gene expression in Lactobacillus strains

Martínez-Fernández, José Alberto; Bravo, Daniel; Peirotén, Ángela; Arqués, Juan Luís; Landete, José María

doi:10.1007/s00253-019-09743-w

Cited by 7 publications

(7 citation statements)

References 24 publications

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“…Here, the behavior of some inducible promoters of genes involved in the transport and the catabolism of carbohydrates and the influence of the presence or absence of cre sites in these promoters were studied by means of a plasmid encoding the evoglow-Pp1 protein as reporter. The use of a flavin mononucleotide-based fluorescent protein presents interesting advantages in some environments and has been successfully used as a reporter (Martínez-Fernández et al 2019;Montenegro-Rodríguez et al 2015). When L. casei BL23 strains transformed with pNZ:gutF.aFP and pNZ:gutF Δ cre .aFP were grown in the presence of sorbitol and ribose, we observed fluorescence induction, being higher in L. casei pNZ:gutF Δcre.…”

Section: Discussionmentioning

confidence: 82%

“…A subsequent study was carried out in order to know what would happen in the event that a cre site was introduced in an induced regulatory system. With this on mind, a bile-induced system previously characterized was used (Martínez-Fernández et al 2019), instead of a carbon source-induced one. In agreement with previous results, L. casei BL23 pNZ:16090.aFP showed induction by 0.5% bile regardless of the presence of glucose.…”

Section: Discussionmentioning

confidence: 99%

“…L. casei BL23 pNZ:malL.aFP and L. casei BL23 pNZ:malL Δcre .aFP were induced by maltose. In P16090 assays, L. casei BL23 pNZ:16090.aFP (Martínez-Fernández et al 2019) and L. casei BL23 pNZ.16090.cre.aFP were supplemented with glucose (0.5%), ribose (0.5%), glucose plus bile salts (both at 0,5%), or ribose plus bile salts (both at 0.5%) or grown in MRS media plus 0.5% bile salts. The same experiments were repeated with different bile salts concentrations (0.25 and 0.05%).…”

Section: Fluorescence Induction Assaysmentioning

confidence: 99%

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Catabolite responsive elements as a strategy for the control of heterologous gene expression in lactobacilli

Langa

Peirotén

Arqués

et al. 2020

Appl Microbiol Biotechnol

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Genes involved in the transport and catabolism of carbohydrates are usually controlled through the binding of the catabolite control protein A (CcpA) to the catabolite-responsive elements (cre) of target genes in Gram-positive bacteria. In this work, we show how the elimination of the cre sites in Lactobacillus casei BL23 promoters induced by sorbitol (PgutF), maltose (PmalL), and myoinositol (PiolT) allowed the induction of gene expression in media supplemented with sorbitol, maltose, and myo-inositol, respectively, even in the presence of glucose. This was studied using plasmids encoding the anaerobic fluorescent protein evoglow-Pp1 as a reporter. In addition, gutF cre site was introduced into a bile inducible promoter (P16090) and into the constitutive promoter of the elongation factor P (PEf-P) of L. casei BL23. The existence of the cre site blocked gene expression in the P16090 inducible promoter in the presence of glucose, while it had no influence on the expression of the PEf-P constitutive one. These results demonstrated that the introduction or elimination of cre sites in inducible promoters allows the control and modification of their heterologous genetic expression, showing how the cre site, the transcriptional regulator, and CcpA interact to control gene expression in inducible genes. Key points• Cre sequences regulate gene expression in inducible promoters in L. casei BL23.• Cre sites do not affect gene expression in constitutive promoters in L. casei BL23.• Cre sequences could control heterologous genic expression in lactobacilli.Keywords catabolite response element . inducible promoter . constitutive promoter . genic expression . Lactobacillus

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Section: Discussionmentioning

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Section: Fluorescence Induction Assaysmentioning

confidence: 99%

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Catabolite responsive elements as a strategy for the control of heterologous gene expression in lactobacilli

Langa

Peirotén

Arqués

et al. 2020

Appl Microbiol Biotechnol

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“…However, the overall protein yield was 60 times lower than the pSIP‐based inducible system, suggesting low expression strength of promoter P MntH2 . Promoter P 16090 isolated from Lactobacillus casei BL23 was tested for its bile stress induction capacity in other bacterial species and was observed to give the highest induction in L. plantarum WCFS1 (Martínez‐Fernández et al, 2019 ). The strain was able to survive at high bile salt concentrations (>0.2% w/v) and showed no leaky expression of the fluorescent reporter protein, evoglow‐Pp1.…”

Section: Inducible Gene Expression Systemsmentioning

confidence: 99%

Expanding the genetic programmability of Lactiplantibacillus plantarum

Blanch‐Asensio,

Dey,

Tadimarri

et al. 2023

Microbial Biotechnology

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Lactobacilli are ubiquitous in nature and symbiotically provide health benefits for countless organisms including humans, animals and plants. They are vital for the fermented food industry and are being extensively explored for healthcare applications. For all these reasons, there is considerable interest in enhancing and controlling their capabilities through the engineering of genetic modules and circuits. One of the most robust and reliable microbial chassis for these synthetic biology applications is the widely used Lactiplantibacillus plantarum species. However, the genetic toolkit needed to advance its applicability remains poorly equipped. This mini‐review highlights the genetic parts that have been discovered to achieve food‐grade recombinant protein production and speculates on lessons learned from these studies for L. plantarum engineering. Furthermore, strategies to identify, create and optimize genetic parts for real‐time regulation of gene expression and enhancement of biosafety are also suggested.

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“…These engineering techniques have been used to activate desirable traits in probiotics such as Lactobacillus casei BL23 , Lb. plantarum WCFS1, and Escherichia coli Nissle 1917; particularly to confer tolerance to specific bile salts, and to produce and secrete antimicrobial peptides to specifically target and kill Enterococcus spp., respectively ( Geldart et al, 2015 , Martínez-Fernández et al, 2019 ).…”

Section: Engineered Probioticsmentioning

confidence: 99%