Folate overproduction in<i>Lactobacillus plantarum</i>WCFS1 causes methotrexate resistance

Wegkamp, Arno; Vos, Willem M. de; Smid, Eddy J.

doi:10.1111/j.1574-6968.2009.01690.x

Cited by 13 publications

(12 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Secondly, specific analyses were performed to determine mechanisms that cause the observed growth rate reduction upon folate overproduction. Previously it was shown that homologous overexpression of the folate gene cluster of L. plantarum results in high folate pools [18]. It was shown that there is 55-fold more folate produced in L. plantarum cultures harboring plasmid pNZ7026 (which carries all genes in the folate biosynthesis pathway) when compared to the control strain carrying plasmid pNZ7021 (empty expression vector) [18].…”

Section: Resultsmentioning

confidence: 99%

“…Previously it was shown that homologous overexpression of the folate gene cluster of L. plantarum results in high folate pools [18]. It was shown that there is 55-fold more folate produced in L. plantarum cultures harboring plasmid pNZ7026 (which carries all genes in the folate biosynthesis pathway) when compared to the control strain carrying plasmid pNZ7021 (empty expression vector) [18]. Using differential metabolomics it was determined whether specific metabolites were more or less abundant in L. plantarum harboring pNZ7026 in comparison to L. plantarum carrying the control plasmid pNZ7021.…”

Section: Resultsmentioning

confidence: 99%

“…Previously, a high folate-producing L. plantarum WCFS1 strain was constructed that produced more than 100-fold increased folate pools, when compared to the control strain. Remarkably, this strain exhibited a 20-25% reduction in growth rate [18]. …”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Physiological responses to folate overproduction in Lactobacillus plantarum WCFS1

et al. 2010

Self Cite

View full text Add to dashboard Cite

BackgroundUsing a functional genomics approach we addressed the impact of folate overproduction on metabolite formation and gene expression in Lactobacillus plantarum WCFS1. We focused specifically on the mechanism that reduces growth rates in folate-overproducing cells.ResultsMetabolite formation and gene expression were determined in a folate-overproducing- and wild-type strain. Differential metabolomics analysis of intracellular metabolite pools indicated that the pool sizes of 18 metabolites differed significantly between these strains. The gene expression profile was determined for both strains in pH-regulated chemostat culture and batch culture. Apart from the expected overexpression of the 6 genes of the folate gene cluster, no other genes were found to be differentially expressed both in continuous and batch cultures. The discrepancy between the low transcriptome and metabolome response and the 25% growth rate reduction of the folate overproducing strain was further investigated. Folate production per se could be ruled out as a contributing factor, since in the absence of folate production the growth rate of the overproducer was also reduced by 25%. The higher metabolic costs for DNA and RNA biosynthesis in the folate overproducing strain were also ruled out. However, it was demonstrated that folate-specific mRNAs and proteins constitute 8% and 4% of the total mRNA and protein pool, respectively.ConclusionFolate overproduction leads to very little change in metabolite levels or overall transcript profile, while at the same time the growth rate is reduced drastically. This shows that Lactobacillus plantarum WCFS1 is unable to respond to this growth rate reduction, most likely because the growth-related transcripts and proteins are diluted by the enormous amount of gratuitous folate-related transcripts and proteins.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Physiological responses to folate overproduction in Lactobacillus plantarum WCFS1

et al. 2010

Self Cite

View full text Add to dashboard Cite

show abstract

“…In this sense, it has been shown that L. plantarum WCSF1, a commensal Lactobacillus strain isolated from human saliva, was shown to possess the folate biosynthesis genes in its genome [45] and that this strain can produce folate in culture media [46]. However, it was shown that only one genetic modification allowed this strain to produce enough of the vitamin to generate a methotrexate (a folate antagonist) resistant phenotype that is observed in high-folate producing strains [47]. Also, it was shown that the riboflavin biosynthesis operon has been shown to be interrupted in L. plantarum WCFS1 genome resulting in its inability to produce riboflavin or other vitamins [45].…”

Section: The Key Role Of Vitamins Produced By Commensal and Probioticmentioning

confidence: 99%

Beneficial effects on host energy metabolism of short-chain fatty acids and vitamins produced by commensal and probiotic bacteria

et al. 2017

View full text Add to dashboard Cite

The aim of this review is to summarize the effect in host energy metabolism of the production of B group vitamins and short chain fatty acids (SCFA) by commensal, food-grade and probiotic bacteria, which are also actors of the mammalian nutrition. The mechanisms of how these microbial end products, produced by these bacterial strains, act on energy metabolism will be discussed. We will show that these vitamins and SCFA producing bacteria could be used as tools to recover energy intakes by either optimizing ATP production from foods or by the fermentation of certain fibers in the gastrointestinal tract (GIT). Original data are also presented in this work where SCFA (acetate, butyrate and propionate) and B group vitamins (riboflavin, folate and thiamine) production was determined for selected probiotic bacteria.

show abstract

“…There is a long history of the application of LAB in a variety of fermentation processes, but especially in the last three decades interest in LAB has significantly increased since many strains, mostly members of the genus Lactobacillus , have health-promoting properties (2–5). In addition, many LAB strains are robust organisms that are able to withstand harsh conditions such as ethanol and stomach acid, which makes them attractive vehicles for the development of industrial producer strains and biotherapeutics, respectively (6–14). Taken together, LAB are therefore the most abundant group of Gram-positive bacteria used in industry and medicine representing a major economic factor.…”

Section: Introductionmentioning

confidence: 99%

CRISPR–Cas9-assisted recombineering in Lactobacillus reuteri

Pijkeren

2014

Nucleic Acids Research

340

245

View full text Add to dashboard Cite

Clustered regularly interspaced palindromic repeats (CRISPRs) and the CRISPR-associated (Cas) nuclease protect bacteria and archeae from foreign DNA by site-specific cleavage of incoming DNA. Type-II CRISPR–Cas systems, such as the Streptococcus pyogenes CRISPR–Cas9 system, can be adapted such that Cas9 can be guided to a user-defined site in the chromosome to introduce double-stranded breaks. Here we have developed and optimized CRISPR–Cas9 function in the lactic acid bacterium Lactobacillus reuteri ATCC PTA 6475. We established proof-of-concept showing that CRISPR–Cas9 selection combined with single-stranded DNA (ssDNA) recombineering is a realistic approach to identify at high efficiencies edited cells in a lactic acid bacterium. We show for three independent targets that subtle changes in the bacterial genome can be recovered at efficiencies ranging from 90 to 100%. By combining CRISPR–Cas9 and recombineering, we successfully applied codon saturation mutagenesis in the L. reuteri chromosome. Also, CRISPR–Cas9 selection is critical to identify low-efficiency events such as oligonucleotide-mediated chromosome deletions. This also means that CRISPR–Cas9 selection will allow identification of recombinant cells in bacteria with low recombineering efficiencies, eliminating the need for ssDNA recombineering optimization procedures. We envision that CRISPR–Cas genome editing has the potential to change the landscape of genome editing in lactic acid bacteria, and other Gram-positive bacteria.

show abstract

Folate overproduction inLactobacillus plantarumWCFS1 causes methotrexate resistance

Cited by 13 publications

References 17 publications

Physiological responses to folate overproduction in Lactobacillus plantarum WCFS1

Physiological responses to folate overproduction in Lactobacillus plantarum WCFS1

Beneficial effects on host energy metabolism of short-chain fatty acids and vitamins produced by commensal and probiotic bacteria

CRISPR–Cas9-assisted recombineering in Lactobacillus reuteri

Contact Info

Product

Resources

About