Identification and Functional Characterization of the<i>Lactococcus lactis</i>CodY-Regulated Branched-Chain Amino Acid Permease BcaP (CtrA)

Hengst, Chris D. den; Groeneveld, Maarten; Kuipers, Oscar P.

doi:10.1128/jb.188.9.3280-3289.2006

Cited by 58 publications

(78 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3A). Although the affinity for other amino acids was not determined, BrnQ appears to be a branched-chain amino acid transporter, as suggested previously (19) and similar to the characterized homologue in Lactobacillus delbrueckii (32). Recently, the yrfD gene, present in strain IL1403 but not in strain MG1363, was proposed to encode the precursor/product exchanger AguD, which would mediate the uptake of extracellular agmatine in exchange for intracellular putrescine, the product of the AgDI pathway (9).…”

Section: Secondary Amino Acid Transporters Of Lactococcus Lactismentioning

confidence: 81%

“…Most of the knowledge still originates from extensive transport studies of whole cells and membrane vesicles derived from wild-type L. lactis strains that were performed in the late 1980s and early 1990s (8,14,16,17,37). Only two amino acid transport systems have since been cloned for expression and functional characterization: the ABC-type glutamate/glutamine transporter GlnPQ, encoded by glnP and glnQ (20), and BcaP, a secondary branched-chain amino acid transporter, encoded by bcaP (19). Here 13 genes of L. lactis MG1363 and 1 gene of L. lactis IL1403 (aguD), with all but one (brnQ) encoding members of the APC family (21) and annotated either as unknown or as amino acid transporters based on homology, were cloned and overexpressed in L. lactis NZ9000.…”

Section: Discussionmentioning

confidence: 99%

“…BrnQ was proposed previously to be a branched-chain amino acid transporter (19,32) but is not related in amino acid sequence to the APC family of transporters. Instead, BrnQ belongs to the LIVCS family of branched-chain amino acid/cation symporters (TC 2.A.26) (21).…”

Section: Secondary Amino Acid Transporters Of Lactococcus Lactismentioning

confidence: 99%

“…The putative amino acid transporter genes, except for ctrA (also known as bcaP), which was well characterized as a branched-chain amino acid transporter previously (19), were expressed in the MG1363-derived strain NZ9000 using the NICE expression system (24). Cells expressing the genes were screened for enhanced uptake by comparing the uptake of 14 amino acids and putrescine (only in case of yrfD) to the uptake by control cells harboring the empty vector pNZ8048.…”

Section: Secondary Amino Acid Transporters Of Lactococcus Lactismentioning

confidence: 99%

“…Only a few amino acid transporters have since been cloned and characterized. For L. lactis, only bcaP and glnPQ were cloned and experimentally shown to encode a branched-chain amino acid transporter (19) and an ATP-driven glutamate/glutamine transporter (20), respectively. Other genes have been annotated as amino acid transporters but mainly on the basis of their homology with other known transporters or their genetic context.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Cloning, Expression, and Functional Characterization of Secondary Amino Acid Transporters of Lactococcus lactis

2013

View full text Add to dashboard Cite

Fourteen genes encoding putative secondary amino acid transporters were identified in the genomes of Lactococcus lactis subsp. cremoris strains MG1363 and SK11 and L. lactis subsp. lactis strains IL1403 and KF147, 12 of which were common to all four strains. Amino acid uptake in L. lactis cells overexpressing the genes revealed transporters specific for histidine, lysine, arginine, agmatine, putrescine, aromatic amino acids, acidic amino acids, serine, and branched-chain amino acids. Substrate specificities were demonstrated by inhibition profiles determined in the presence of excesses of the other amino acids. Four knockout mutants, lacking the lysine transporter LysP, the histidine transporter HisP (formerly LysQ), the acidic amino acid transporter AcaP (YlcA), or the aromatic amino acid transporter FywP (YsjA), were constructed. The LysP, HisP, and FywP deletion mutants showed drastically decreased rates of uptake of the corresponding substrates at low concentrations. The same was observed for the AcaP mutant with aspartate but not with glutamate. In rich M17 medium, the deletion of none of the transporters affected growth. In contrast, the deletion of the HisP, AcaP, and FywP transporters did affect growth in a defined medium with free amino acids as the sole amino acid source. HisP was essential at low histidine concentrations, and AcaP was essential in the absence of glutamine. FywP appeared to play a role in retaining intracellularly synthesized aromatic amino acids when these were not added to the medium. Finally, HisP, AcaP, and FywP did not play a role in the excretion of accumulated histidine, glutamate, or phenylalanine, respectively, indicating the involvement of other transporters.

show abstract

Section: Secondary Amino Acid Transporters Of Lactococcus Lactismentioning

confidence: 81%

Section: Discussionmentioning

confidence: 99%

Section: Secondary Amino Acid Transporters Of Lactococcus Lactismentioning

confidence: 99%

Section: Secondary Amino Acid Transporters Of Lactococcus Lactismentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Cloning, Expression, and Functional Characterization of Secondary Amino Acid Transporters of Lactococcus lactis

2013

View full text Add to dashboard Cite

show abstract

Nutritional requirements and media development for Lactococcus lactis IL1403

Aller

Adamberg

Timarova

et al. 2014

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Lactic acid bacteria are extensively used in food technology and for the production of various compounds, but they are fastidious in nutrient requirements. In order to elucidate the role of each component precisely, defined multicomponent media are required. This study focuses on determining nutrient auxotrophies and minimizing media components (amino acids, vitamins, metal ions, buffers and additional compounds) for the cultivation of Lactococcus lactis subsp. lactis IL1403, using microtitre plates and test tubes. It was shown that glutamine and asparagine were the most important media components for achieving higher biomass yields while the branched-chain amino acids were necessary to increase specific growth rate. The amino acid and glucose ratio was reduced to achieve minimal residual concentration of amino acids in the medium after the growth of cells, whereas the specific growth rate and biomass yield of cells were not considerably affected. As the percentage of each consumed amino acid compared to initial amount is larger than measurement error, these optimized media are important for achieving more precise data about amino acid utilization and metabolism.

show abstract

Microbial methionine transporters and biotechnological applications

Mohany

Totti

Naylor

et al. 2021

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Methionine (Met) is an essential amino acid with commercial value in animal feed, human nutrition, and as a chemical precursor. Microbial production of Met has seen intensive investigation towards a more sustainable alternative to the chemical synthesis that currently meets the global Met demand. Indeed, efficient Met biosynthesis has been achieved in genetically modified bacteria that harbor engineered enzymes and streamlined metabolic pathways. Very recently, the export of Met as the final step during its fermentative production has been studied and optimized, primarily through identification and expression of microbial Met efflux transporters. In this mini-review, we summarize the current knowledge on four families of Met export and import transporters that have been harnessed for the production of Met and other valuable biomolecules. These families are discussed with respect to their function, gene regulation, and biotechnological applications. We cover methods for identification and characterization of Met transporters as the basis for the further engineering of these proteins and for exploration of other solute carrier families. The available arsenal of Met transporters from different species and protein families provides blueprints not only for fermentative production but also synthetic biology systems, such as molecular sensors and cell-cell communication systems. Key points • Sustainable production of methionine (Met) using microbes is actively explored. • Met transporters of four families increase production yield and specificity. • Further applications include other biosynthetic pathways and synthetic biology.

show abstract

Identification and Functional Characterization of theLactococcus lactisCodY-Regulated Branched-Chain Amino Acid Permease BcaP (CtrA)

Cited by 58 publications

References 67 publications

Cloning, Expression, and Functional Characterization of Secondary Amino Acid Transporters of Lactococcus lactis

Cloning, Expression, and Functional Characterization of Secondary Amino Acid Transporters of Lactococcus lactis

Nutritional requirements and media development for Lactococcus lactis IL1403

Microbial methionine transporters and biotechnological applications

Contact Info

Product

Resources

About