Biotin uptake: influx, efflux and countertransport in Escherichia coli K12

Piffeteau, Annie; Gaudry, Michel

doi:10.1016/0005-2736(85)90395-5

Cited by 36 publications

(22 citation statements)

References 10 publications

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“…However, since the bioY disruption has satisfied our purpose, those additional experiments have not yet been carried out. In this study, the bioY disruptant still grew under the biotin excess conditions, but this is probably due to the entry of biotin into the cells by passive diffusion, as was observed in E. coli (44).…”

Section: Discussionmentioning

confidence: 63%

Development of Biotin-Prototrophic and -Hyperauxotrophic Corynebacterium glutamicum Strains

Ikeda

Miyamoto

Mutoh

et al. 2013

Appl Environ Microbiol

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bTo develop the infrastructure for biotin production through naturally biotin-auxotrophic Corynebacterium glutamicum, we attempted to engineer the organism into a biotin prototroph and a biotin hyperauxotroph. To confer biotin prototrophy on the organism, the cotranscribed bioBF genes of Escherichia coli were introduced into the C. glutamicum genome, which originally lacked the bioF gene. The resulting strain still required biotin for growth, but it could be replaced by exogenous pimelic acid, a source of the biotin precursor pimelate thioester linked to either coenzyme A (CoA) or acyl carrier protein (ACP). To bridge the gap between the pimelate thioester and its dedicated precursor acyl-CoA (or -ACP), the bioI gene of Bacillus subtilis, which encoded a P450 protein that cleaves a carbon-carbon bond of an acyl-ACP to generate pimeloyl-ACP, was further expressed in the engineered strain by using a plasmid system. This resulted in a biotin prototroph that is capable of the de novo synthesis of biotin. On the other hand, the bioY gene responsible for biotin uptake was disrupted in wild-type C. glutamicum. Whereas the wildtype strain required approximately 1 g of biotin per liter for normal growth, the bioY disruptant (⌬bioY) required approximately 1 mg of biotin per liter, almost 3 orders of magnitude higher than the wild-type level. The ⌬bioY strain showed a similar high requirement for the precursor dethiobiotin, a substrate for bioB-encoded biotin synthase. To eliminate the dependency on dethiobiotin, the bioB gene was further disrupted in both the wild-type strain and the ⌬bioY strain. By selectively using the resulting two strains (⌬bioB and ⌬bioBY) as indicator strains, we developed a practical biotin bioassay system that can quantify biotin in the seven-digit range, from approximately 0.1 g to 1 g per liter. This bioassay proved that the engineered biotin prototroph of C. glutamicum produced biotin directly from glucose, albeit at a marginally detectable level (approximately 0.3 g per liter).

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Section: Discussionmentioning

confidence: 63%

Development of Biotin-Prototrophic and -Hyperauxotrophic Corynebacterium glutamicum Strains

Ikeda

Miyamoto

Mutoh

et al. 2013

Appl Environ Microbiol

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“…Figure 3 shows that avidin could competitively inhibit the uptake of biotinylated peptides in E. coli but that the use of another similarly sized protein, bovine serum albumin, which is routinely used in in vitro studies, had no effect. It has been shown that biotin uptake is blocked by the protonophore CCCP, which disrupts membrane potential in E. coli (34,35). If the uptake of biotinylated peptides was due to the biotin transport system, then CCCP would be expected to block the uptake of biotinylated peptides.…”

Section: Resultsmentioning

confidence: 99%

“…E. coli readily imports the vitamin when it is available and concomitantly represses biotin synthesis. Biotin uptake is specific and energy dependent and can accumulate against a concentration gradient (34,35,36). Maximum uptake is observed during the exponential growth phase (34), and glucose has been shown to increase biotin uptake slightly.…”

Section: Vol 71 2005 Biotinylation Facilitates the Uptake Of Large mentioning

confidence: 99%

Biotinylation Facilitates the Uptake of Large Peptides by Escherichia coli and Other Gram-Negative Bacteria

Walker¹,

Altman

2005

Appl Environ Microbiol

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Gram-negative bacteria such as Escherichia coli can normally only take up small peptides less than 650 Da, or five to six amino acids, in size. We have found that biotinylated peptides up to 31 amino acids in length can be taken up by E. coli and that uptake is dependent on the biotin transporter. Uptake could be competitively inhibited by free biotin or avidin and blocked by the protonophore carbonyl m-chlorophenylhydrazone and was abolished in E. coli mutants that lacked the biotin transporter. Biotinylated peptides could be used to supplement the growth of a biotin auxotroph, and the transported peptides were shown to be localized to the cytoplasm in cell fractionation experiments. The uptake of biotinylated peptides was also demonstrated for two other gram-negative bacteria, Salmonella enterica serovar Typhimurium and Pseudomonas aeruginosa. This finding may make it possible to create new peptide antibiotics that can be used against gram-negative pathogens. Researchers have used various moieties to cause the illicit transport of compounds in bacteria, and this study demonstrates the illicit transport of the largest known compound to date.The outer membrane of gram-negative bacteria functions as a molecular sieve and allows only very small molecules to passively diffuse into the cell. Porins in the outer membrane allow the transport of larger molecules and may be specific or nonspecific in their molecular recognition. Nonspecific porins such as OmpF, OmpC, and PhoE allow the rapid passage of hydrophilic molecules (27,28). Other porins allow the transport of specific molecules. The peptide permeases, for example, have a specificity for oligopeptides. The uptake of oligopeptides is dependent upon size, hydrophobicity, and charge (5,26,32).It is well documented that Escherichia coli cannot take up large peptides and that the size exclusion limit for porin-mediated peptide transport is 650 Da or the size of a penta-or hexapeptide (31, 33). The size exclusion limit for peptide uptake in other gram-negative organisms such as Salmonella enterica serovar Typhimurium has also been determined and found to be similar to that in E. coli (31,33). In contrast to gram-negative bacteria, gram-positive bacteria can transport much larger peptides. For example, Lactococcus lactis has been shown to take up peptides more than 18 residues in length or 2,140 Da in size (10) while Bacillus megaterium can transport molecules up to 10,000 Da in size (40).This study provides evidence that large biotinylated peptides can be readily transported into gram-negative bacteria such as E. coli. While conducting an in vivo screen for randomly encoded peptides that could inhibit the growth of Staphylococcus aureus, we performed a test to confirm that potential peptides resulting from the screen would be readily taken up, as expected, by this gram-positive organism. A biotinylated 10-amino-acid peptide was added extracellularly to growing cultures of S. aureus and an E. coli control, which should not have been able to take up the 1,534-Da peptide...

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“…Here, conserved hydrophobic residues in helices 2 and 3 of the N-terminal domain of the EcfS subunit, containing the highly conserved AφφφA signature (where φ is mostly a hydrophobic residue) [30], form hydrophobic and hydrogen bonding interactions with conserved hydrophobic residues in trans-membrane helices 1-4 and cytoplasmic helix 6 of EcfT [27,39]. Hydrophobic residues in helix 6 and loops L3 and L5 of EcfS also participate in the interactions with EcfT [27], enforcing the predicted role of EcfT as a stabiliser of the complex [26,27,39,42,53]. This structural data also provides a molecular explanation as to how EcfT of Class II transporters can interact with multiple EcfS components.…”

Section: Intersubunit Interactionsmentioning

confidence: 79%

Mechanisms of Biotin Transport

Polyak¹

2015

Biochem Anal Biochem

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Biotin uptake: influx, efflux and countertransport in Escherichia coli K12

Cited by 36 publications

References 10 publications

Development of Biotin-Prototrophic and -Hyperauxotrophic Corynebacterium glutamicum Strains

Development of Biotin-Prototrophic and -Hyperauxotrophic Corynebacterium glutamicum Strains

Biotinylation Facilitates the Uptake of Large Peptides by Escherichia coli and Other Gram-Negative Bacteria

Mechanisms of Biotin Transport

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