Lactate Utilization Is Regulated by the FadR-Type Regulator LldR in Pseudomonas aeruginosa

Gao, Chao; Hu, Chunhui; Zheng, Zhaojuan; Ma, Cuiqing; Jiang, Tianyi; Dou, Peipei; Zhang, Wen; Che, Bin; Wang, Yujiao; Lv, Min; Xu, Ping

doi:10.1128/jb.06579-11

Cited by 52 publications

(76 citation statements)

References 34 publications

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“…However, the MIC M s for 98% of the P. aeruginosa strains with citric and lactic acids were at pH 4.53 and pH 5.28, respectively [35]. The result obtained for P. aeruginosa [35] may well be different since P. aeruginosa is known to utilize lactate [45,46]. The 100% inhibition range for all four undissociated organic acids, acetic, citric, lactic and propionic for the 145 Salmonella strains extends from 2.29 mM undissociated citric acid to 19.0 mM undissociated acetic acid.…”

Section: Beier Et Almentioning

confidence: 64%

Interactions of Organic Acids with Salmonella Strains from Feedlot Water-Sprinkled Cattle

Beier¹,

Callaway²,

Andrews³

et al. 2017

J Food Chem Nanotechnol

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Section: Beier Et Almentioning

confidence: 64%

Interactions of Organic Acids with Salmonella Strains from Feedlot Water-Sprinkled Cattle

Beier¹,

Callaway²,

Andrews³

et al. 2017

J Food Chem Nanotechnol

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“…Therefore, bacterial genetic operons represent a rich source for designing new whole‐cell biosensors for application in industrial bioprocessing. In particular, operons to utilize lactate as a carbon source have been described in Escherichia coli (Aguilera et al, 2008), Cornyebacterium (Gao et al, 2008), Pseudomonas (Gao et al, 2012), and Bacillus subtilis (Chai et al, 2009). Any of these can be used to design a whole‐cell biosensor by co‐opting the existing genetic elements to sense lactate and activate gene expression and using them to express a reporter protein in response to lactate induction.…”

Section: Introductionmentioning

confidence: 99%

Whole‐cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production

Goers

Ainsworth

Goey

et al. 2017

Biotech & Bioengineering

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Many high‐value added recombinant proteins, such as therapeutic glycoproteins, are produced using mammalian cell cultures. In order to optimize the productivity of these cultures it is important to monitor cellular metabolism, for example the utilization of nutrients and the accumulation of metabolic waste products. One metabolic waste product of interest is lactic acid (lactate), overaccumulation of which can decrease cellular growth and protein production. Current methods for the detection of lactate are limited in terms of cost, sensitivity, and robustness. Therefore, we developed a whole‐cell Escherichia coli lactate biosensor based on the lldPRD operon and successfully used it to monitor lactate concentration in mammalian cell cultures. Using real samples and analytical validation we demonstrate that our biosensor can be used for absolute quantification of metabolites in complex samples with high accuracy, sensitivity, and robustness. Importantly, our whole‐cell biosensor was able to detect lactate at concentrations more than two orders of magnitude lower than the industry standard method, making it useful for monitoring lactate concentrations in early phase culture. Given the importance of lactate in a variety of both industrial and clinical contexts we anticipate that our whole‐cell biosensor can be used to address a range of interesting biological questions. It also serves as a blueprint for how to capitalize on the wealth of genetic operons for metabolite sensing available in nature for the development of other whole‐cell biosensors. Biotechnol. Bioeng. 2017;114: 1290–1300. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

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“…The L-lactateresponsive regulator LldR negatively regulates expression of the lldPRD operon through binding to two operator sites containing inverted repeats with the putative consensus sequence 59-AATTGGCA-N 1 -TGCCAATT-39 (Aguilera et al, 2008). The lldPDE operon of Pseudomonas aeruginosa XMG comprises genes encoding the lactate permease LldP, the L-lactate dehydrogenase LldD and the D-lactate dehydrogenase LldE (Kemp, 1972;Gao et al, 2012a). The lldPDE operon is located adjacent to the lldR gene, which codes for a GntR-type regulator LldR.…”

Section: Introductionmentioning

confidence: 99%

“…The lldPDE operon is located adjacent to the lldR gene, which codes for a GntR-type regulator LldR. Both L-lactate and D-lactate can interfere with the binding of the LldR repressor to its operator, which contains an inverted repeat sequence 59-TGGT-N 3 -ACCA-39 (Gao et al, 2012a). In the non-pathogenic Gram-positive bacterium Corynebacterium glutamicum ATCC 13032, the L-lactate utilization operon lldPD comprises the L-lactate permease gene lldP and the L-lactate dehydrogenase gene lldD (Stansen et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Transcriptional regulation of the l-lactate permease gene lutP by the LutR repressor of Bacillus subtilis RO-NN-1

Chiu¹,

Lin²,

Shaw³

2014

Microbiology

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The Bacillus subtilis lutABC operon encodes three iron-sulfur-containing proteins required for Llactate utilization and involved in biofilm formation. The transcriptional regulator LutR of the GntR family negatively controls lutABC expression. The lutP gene, which is situated immediately upstream of lutR, encodes an L-lactate permease. Here, we show that lutP expression can be strongly induced by L-lactate and is subject to partial catabolite repression by glucose. Disruption of the lutR gene led to a strong derepression of lutP and no further induction by L-lactate, suggesting that the LutR repressor can also negatively control lutP expression. Electrophoretic mobility shift assay revealed a LutR-binding site located downstream of the promoter of lutA or lutP and containing a consensus inverted repeat sequence 59-TCATC-N 1 -GATGA-39. Reporter gene analysis showed that deletion of each LutR-binding site caused a strong derepression of lutA or lutP. These results indicated that these two LutR-binding sites can function as operators in vivo. Moreover, deletion analysis identified a DNA segment upstream of the lutP promoter to be important for lutP expression. In contrast to the truncated LutR of laboratory strains 168 and PY79, the full-length LutR of the undomesticated strain RO-NN-1, and probably many other B. subtilis strains, can directly and negatively regulate lutP transcription. The absence or presence of the N-terminal 21 aa of the full-length LutR, which encompass a small part of the predicted winged helix-turn-helix DNA-binding motif, may probably alter the DNA-binding specificity or affinity of LutR.

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Lactate Utilization Is Regulated by the FadR-Type Regulator LldR in Pseudomonas aeruginosa

Cited by 52 publications

References 34 publications

Interactions of Organic Acids with Salmonella Strains from Feedlot Water-Sprinkled Cattle

Interactions of Organic Acids with Salmonella Strains from Feedlot Water-Sprinkled Cattle

Whole‐cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production

Transcriptional regulation of the l-lactate permease gene lutP by the LutR repressor of Bacillus subtilis RO-NN-1

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